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+arg_1	round_1	ann_1	arg_2	round_2	ann_2	annotation_name	is_attacks
+I have tried to address your principle concern by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	As a separate recommendation, the paper needs smoother transitions between topics on pages 2-7. The authors should add one or two lines smoothing the flow for the reader, rather than relying on subject headings.	1	2	admsci5030125_boyarkin	1
+I have tried to improve the information density of the paper from being to verbiose.	2	1	The paper is on the one hand quite verbose, but with respect to the peer review incomplete. Possible risks in peer review are many more than the ones listed in Table 3.	1	2	admsci5030125_boyarkin	1
+I have added the results of the stakeholder analysis to improve the communication our view of the PRP.	2	1	The paper can become stronger if it limits itself to the Peer Review Ring Incident and will look at the various aspects of that scenario including risk reducing measures.	1	2	admsci5030125_boyarkin	1
+I have tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	It didn't become clear to me as a reader where the specific results of the 13 steps of the CIRA application are and where these are described. What are the conclusions about the root cause? Or are there multiple root causes? And what definition of root cause is used?	1	2	admsci5030125_boyarkin	1
+I have elaborated on all the areas you commented, and clarified the CIRA methodology and hopefully my contribution.	2	1	there is not enough explanation about the CIRA and the person(s) who developed this method. Also there is not enough clarification and elaboration on the differences of your work on CIRA and your customizations comparing to the original methodology.	1	2	admsci5030125_boyarkin	1
+Thank you for comments on the grammar issues, these are now fixed. Abbrevation issues should be fixed.	2	1	There are some typos, punctuations and grammatical errors in the paper,	1	2	admsci5030125_boyarkin	1
+I have also tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	There are some consistency issues in your writing. This does not look professional in scientific papers.	1	2	admsci5030125_boyarkin	1
+I have included all your suggested changes together with a major spellcheck.	4	1	There is a need for some editing; e.g. grammatical and punctuation corrections, elements in the text.	3	2	admsci5030125_boyarkin	1
+I have tried to address your principle concern by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	Given the authors stated goal of promoting the CIRA technique and approach to demonstrating its uses, I would recommend re-formatting the paper as a pedagogical tool. Rather than claiming to validate the method, focus on a detailed step by step examination of its implementation using a known outcome.	1	2	admsci5030125_makarova	1
+I have tried to improve the information density of the paper from being to verbiose.	2	1	The paper is on the one hand quite verbose, but with respect to the peer review incomplete.	1	2	admsci5030125_makarova	1
+I have added the results of the stakeholder analysis to improve the communication our view of the PRP.	2	1	The paper can become stronger if it limits itself to the Peer Review Ring Incident and will look at the various aspects of that scenario including risk reducing measures.	1	2	admsci5030125_makarova	1
+I have tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	What are the conclusions about the root cause? Or are there multiple root causes? And what definition of root cause is used?	1	2	admsci5030125_makarova	1
+I have elaborated on all the areas you commented, and clarified the CIRA methodology and hopefully my contribution.	2	1	Also there is not enough clarification and elaboration on the differences of your work on CIRA and your customizations comparing to the original methodology.	1	2	admsci5030125_makarova	1
+I have included all your suggested changes together with a major spellcheck.	4	1	There is a need for some editing; e.g. grammatical and punctuation corrections, elements in the text.	3	2	admsci5030125_makarova	1
+Agree that it should not be an issue. Therefore, I recalibrated the piece to note that co-operatives are not third sector organizations, but instead have similar features that could allow the institutional analyst fertile ground for innovations in governance.	2	1	the third sector, its definition and description is missing. I would not have thought of this as a major issue unless Evers and Laville (2004) The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector. Maybe taken for granted here? The least we should expect is a definition.	1	2	admsci5030148_devetyaro	1
+I agree that this is not a case study; I made the mistake in haste. I have therefore changed the paper from a case study to a discussion paper. Additionally, I note in the paper that I perform an archival analysis of media accounts in an effort to demonstrate how the utilization of the ODPs for analysizing secondary data may better position the analyst to perform rigorous, interogative fieldwork.	2	1	the case description and analysis is based only on media accounts - no actual first hand information to inform the analysis. I find this less than satisfying. It should be possible to interview a few key individuals about the case to make sure the medis picture is corrected where this is needed. Media does not always get it right. There is not enough evidence to prove anything about CEC.	1	2	admsci5030148_devetyaro	1
+See the DISCUSSION and CONCLUSION sections, where I have added additional analyses.	2	1	I also have problems with the depth of the discussion, and I feel the conclusion does not match the promises of the abstract.	1	2	admsci5030148_devetyaro	1
+Corrected and accounted for.	2	1	Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders)	1	2	admsci5030148_devetyaro	1
+I do not believe this is a necessary reference as I cite the International Co-operative Alliance principles, and the Rochdale Principles are somewhat commonly understood.	2	1	Bottom of page 9: Missing reference to Co-operative principles by Rochdale Society.	1	2	admsci5030148_devetyaro	1
+Accounted for.	2	1	USA context concerning cooperatives (upper half page 10) should be highlighted even more for readers to be able to compare to their own context (for instance in Europe)	1	2	admsci5030148_devetyaro	1
+Addressed in the text.	2	1	The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?	1	2	admsci5030148_devetyaro	1
+Corrected.	2	1	are references 27 and 33 used in the text? (could not find them) Good luck in further research and writing on this subject.	1	2	admsci5030148_devetyaro	1
+I have changed the headings for these sections to better reflect the content. They read as follows: 2.	4	1	The literature part is improved by changing the headline, putting it in the right setting. The change illustrates how headlines can change the apperance of the text contents. However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles? If they are - the connection need explainnig. If they are not, the main headling Chapter 2 should be reformulated again.	3	2	admsci5030148_devetyaro	1
+This is general knowledge, thereby not necessitating a reference. If the reviewers disagree, I would add a reference.	4	1	I still miss a source reference at the bottom of page 9 (Rochdale Society).	3	2	admsci5030148_devetyaro	1
+I have added commentary about moral hazard and information asymetries.	4	1	On top of page 11 there is still no examples of adversarial effects removed by co-operative.	3	2	admsci5030148_devetyaro	1
+Agreed. Agreed. I have added additional elaboration.	4	1	The comments added in the beginning of Chapter 3 on methodology are important and improves the text a lot. The author openly acknowledges that the basis for a complete picture is lacking. I do not expect the complete picture, but should have comments on limitations and to what degree the conclusions and learnings are possible to generalize.	3	2	admsci5030148_devetyaro	1
+Commentary added, nothing the need for additiona interogation.	4	1	"Conclusion: middle of page 16: ""we cannot know the motivations behind.."" Have you tried to talk to them? I believe it is possible to get to know."	3	2	admsci5030148_devetyaro	1
+Commentary added for further clarity.	4	1	"Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"	3	2	admsci5030148_devetyaro	1
+"I agree with your points about the over-emphasis on principle-agent types of governance arrangements. The Cornforth (2004) article reoriented my thinking (thank you). I recognized that it was not ""P-A"" I was critiquing, but the unitarist tradition. That approach helped with definitional rigor"	2	1	Furthermore, there is no discussion of principal-agent literature in a third sector context (see, for example, Jegers, 2009), which is quite surprising since the proposition to go beyond the traditional principal-agent literature is one of the main ideas of the manuscript.	1	2	admsci5030148_devetyaro	1
+"I believe the recalibration of the core critique from ""P-A"" to unitarism addresses the commentary, here."	2	1	Furthermore, attempts have already been made to broaden the principal-agent approach in nonprofit organizations (Ben-Ner et al., 2012; Van Puyvelde et al., 2012, Coule, 2015), leading me to the question what this manuscript actually adds to the literature.	1	2	admsci5030148_devetyaro	1
+"A further recalibration. I adusted the commentary to address ""pluralistic"" governance as opposed to stakeholder governance."	2	1	In the introduction, the author also mentions underdeveloped modes of stakeholder governance in the third sector. I do not agree with this view. Several stakeholder governance frameworks for third sector organizations have already been developed in the literature (Young, 2011; Van Puyvelde et al., 2012; Wellens and Jegers, 2014).	1	2	admsci5030148_devetyaro	1
+I editted the paper for consistenct, in line with your commentary.	2	1	The author uses “stakeholder-controlled firm”, “third sector organizations”, “nonprofit organizations”, “social enterprise”, and “co-operative” without clearly defining these types and mentioning which nonprofit type is actually the focus of the study.	1	2	admsci5030148_devetyaro	1
+The definition used is from the official representative body, the International Co-operative Alliance.	2	1	I am also not convinced by the definition of co-operatives mentioned in the paper.	1	2	admsci5030148_devetyaro	1
+Acknowledged. First, the paper is no longer listed as a case study, but as a discussion piece. Second, I note in the paper that I am not making the claim that the co-operative model is part of the third sector.	2	1	Furthermore, given that the world of third sector organizations is very rich and heterogeneous, one should be careful by discussing third sector organizations in general, especially since the case study seems to focus on co-operative enterprises. Instead of being vague and using a mixed terminology, the manuscript should clearly distinguish which nonprofit type is the subject of the paper, and be very careful in generalizing the findings of this study to third sector governance, which mistakenly happens in the conclusion (p. 15). Given the case study, I would focus on the governance of co-operatives	1	2	admsci5030148_devetyaro	1
+"Acknowledged. The ""Literature Review"" section now falls under a more accurate heading of ""Introducing the Ostrom Design Principles."""	2	1	The literature review is completely missing.	1	2	admsci5030148_devetyaro	1
+I believe that this then addresses the issues that fall in line with your critiques about the theoretical foundation.	2	1	The literature review is completely missing.	1	2	admsci5030148_devetyaro	1
+As per your suggestions, I did focus on the co-operative model I believe that Cornforth’s paper (2004) might be a good starting point.	2	1	Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.	1	2	admsci5030148_devetyaro	1
+Very helpful! Thank you!	2	1	I believe that Cornforth’s paper (2004) might be a good starting point.	1	2	admsci5030148_devetyaro	1
+Note that the paper no longer claims this to be a case study, and instead positions itself as a discussion paper, performing a secondary archival analysis of media accounts.	2	1	In addition, a more extensive literature review and comparison with previous principal-agent literature and stakeholder governance models (especially Van Puyvelde et al., 2012 and Coule, 2015) may give the manuscript a more solid theoretical foundation. As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.	1	2	admsci5030148_devetyaro	1
+Thank you.	2	1	The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.	1	2	admsci5030148_devetyaro	1
+I agree with your critique. I have adjusted accordingly. I did this by separating the co-operative model from the third sector, and instead noted that there exists commonalities which allows for comparative learning and innovations in governance.	2	1	The characterisation of third sector governance is too crude. For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors. The comments at the start of the literature review might apply to foundations and charities, but are less true of association and mutuals (let alone cooperatives). Also, where there are accounting regulations (like SORP), there are mechanism that keep associations and charities accountable to their donors.	1	2	admsci5030148_devetyaro	1
+Note that throughout the document I incorporate Coule's unitary-pluralist dimensions.	2	1	Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.	1	2	admsci5030148_devetyaro	1
+Agreed and accounted for.	2	1	Secondly, I just can't agree that third sector governance is derived from for-profit principles. It is - mostly - derived from philanthropic principles (or mutual principles inside the social economy). There is, however, some common practices, but not for the reasons described. Just as the wealthy entrepreneur seeks to control the private enterprise through shareholdings, so the wealthy philanthropist seeks to control the non-profit enterprise through trust law. In both cases, Principle-Agent assumptions apply, but not because both are based on for-profit assumptions. These apply in both cases because funding is sought from wealthy providers with the power to frame laws that require their investees to use the money for the purposes stimpulated by them. In the social economy (associations, mutuals and cooperatives) the legal structures and accountabilities are different (and are captured well in the paper). Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal structures and accountability mechanisms reflect this.	1	2	admsci5030148_devetyaro	1
+Agreed. I misrepresented the analysis. It is not a case study, but instead an cursory archival analysis and exploration, utilizing secondary sources.	2	1	This is weak, perhaps too weak for an academic paper. Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable. The use of media sources, while not ideal, might be adequate if the author(s) can show systematic and comprehensive collection of them. However, the credibility of the paper and findings would be enormously enhanced is some primary interview data could be collected and presented. Is there a way to interview members who participated in action (or access sources written by them)?	1	2	admsci5030148_devetyaro	1
+Agreed. The Analysis, Discussiom, and Conclusion sections are more fleshed out.	2	1	The framework for this section exists but feels a bit superficial and under-developed. I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).	1	2	admsci5030148_devetyaro	1
+Agreed, and accounted for.	2	1	Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification. Stick to commenting on the usefulness and value of ODPs, and draw out any theoretical contributions regarding the design principles themselves, or their value as a theoretical perspective for governance research. Some well grounded comments on their use in other governance research are merited right at the end.	1	2	admsci5030148_devetyaro	1
+I hope that the revisions in the abstract an introduction better clarify the purpose.	2	1	I do not understand the purpose of the paper.	1	2	admsci5030148_devetyaro	1
+Agreed. I accounted for this throughout the document. Terminology is used in a definitionally consistent manner.	2	1	The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.	1	2	admsci5030148_devetyaro	1
+Note that I adjusted the paper, and recalibrated it from a case study, to a discussion paper that utilizes an archival analysis of secondary sources. I do this in order to demonstrate how the Ostrom Design Principles may be used by the researcher to better contextualize the subject being studied so they may enter the field prepared to deeply interrogate.	2	1	The style is more journalistic and free opinion than rigourous research.	1	2	admsci5030148_devetyaro	1
+Noted and accounted for throughout the document.	2	1	Public enterprises, third sector cooperatives are all mixedup.	1	2	admsci5030148_devetyaro	1
+Acknowledged. I cleaned and honed the Introduction, and added a more robust Conclusion.	2	1	The conceptual framework is not clearly presented and I do not see what are the contributions except some free statements at the end not really related to the analysis.	1	2	admsci5030148_devetyaro	1
+Therefore, I recalibrated the piece to note that co-operatives are not third sector organizations, but instead have similar features that could allow the institutional analyst fertile ground for innovations in governance.	2	1	the third sector, its definition and description is missing. The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector.	1	2	admsci5030148_makarova	1
+I have therefore changed the paper from a case study to a discussion paper. Additionally, I note in the paper that I perform an archival analysis of media accounts in an effort to demonstrate how the utilization of the ODPs for analysizing secondary data may better position the analyst to perform rigorous, interogative fieldwork.	2	1	the case description and analysis is based only on media accounts - no actual first hand information to inform the analysis.	1	2	admsci5030148_makarova	1
+See the DISCUSSION and CONCLUSION sections, where I have added additional analyses.	2	1	I also have problems with the depth of the discussion, and I feel the conclusion does not match the promises of the abstract.	1	2	admsci5030148_makarova	1
+Corrected and accounted for.	2	1	Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders)	1	2	admsci5030148_makarova	1
+I do not believe this is a necessary reference as I cite the International Co-operative Alliance principles, and the Rochdale Principles are somewhat commonly understood.	2	1	Bottom of page 9: Missing reference to Co-operative principles by Rochdale Society.	1	2	admsci5030148_makarova	1
+Accounted for.	2	1	USA context concerning cooperatives (upper half page 10) should be highlighted even more for readers to be able to compare to their own context (for instance in Europe)	1	2	admsci5030148_makarova	1
+Addressed in the text.	2	1	The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?	1	2	admsci5030148_makarova	1
+Corrected.	2	1	are references 27 and 33 used in the text? (could not find them) Good luck in further research and writing on this subject.	1	2	admsci5030148_makarova	1
+I have changed the headings for these sections to better reflect the content.	4	1	However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles? If they are - the connection need explainnig. If they are not, the main headling Chapter 2 should be reformulated again.	3	2	admsci5030148_makarova	1
+This is general knowledge, thereby not necessitating a reference. If the reviewers disagree, I would add a reference.	4	1	I still miss a source reference at the bottom of page 9 (Rochdale Society).	3	2	admsci5030148_makarova	1
+I have added commentary about moral hazard and information asymetries.	4	1	On top of page 11 there is still no examples of adversarial effects removed by co-operative.	3	2	admsci5030148_makarova	1
+Agreed. Agreed. I have added additional elaboration.	4	1	The author openly acknowledges that the basis for a complete picture is lacking. I do not expect the complete picture, but should have comments on limitations and to what degree the conclusions and learnings are possible to generalize.	3	2	admsci5030148_makarova	1
+Commentary added, nothing the need for additiona interogation.	4	1	"Conclusion: middle of page 16: ""we cannot know the motivations behind.."" Have you tried to talk to them? I believe it is possible to get to know."	3	2	admsci5030148_makarova	1
+Commentary added for further clarity.	4	1	"Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"	3	2	admsci5030148_makarova	1
+I agree with your points about the over-emphasis on principle-agent types of governance arrangements.	2	1	The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation. Similarly, in the introduction, it is stated that governance research on third sector organizations predominantly fixates analysts on the board-management dynamic.	1	2	admsci5030148_makarova	1
+"I believe the recalibration of the core critique from ""P-A"" to unitarism addresses the commentary, here."	2	1	In the abstract, the author also states that the over-reliance on the principal-agent model introduces two challenges. However, a solid foundation for this argument is missing.	1	2	admsci5030148_makarova	1
+"A further recalibration. I adusted the commentary to address ""pluralistic"" governance as opposed to stakeholder governance."	2	1	In the introduction, the author also mentions underdeveloped modes of stakeholder governance in the third sector. I do not agree with this view. Several stakeholder governance frameworks for third sector organizations have already been developed in the literature (Young, 2011; Van Puyvelde et al., 2012; Wellens and Jegers, 2014).	1	2	admsci5030148_makarova	1
+I editted the paper for consistenct, in line with your commentary.	2	1	The author uses “stakeholder-controlled firm”, “third sector organizations”, “nonprofit organizations”, “social enterprise”, and “co-operative” without clearly defining these types and mentioning which nonprofit type is actually the focus of the study.	1	2	admsci5030148_makarova	1
+The definition used is from the official representative body, the International Co-operative Alliance.	2	1	I am also not convinced by the definition of co-operatives mentioned in the paper.	1	2	admsci5030148_makarova	1
+As per your suggestions, I did focus on the co-operative model I believe that Cornforth’s paper (2004) might be a good starting point.	2	1	Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.	1	2	admsci5030148_makarova	1
+I agree with your critique. I have adjusted accordingly.	2	1	The characterisation of third sector governance is too crude.	1	2	admsci5030148_makarova	1
+Note that throughout the document I incorporate Coule's unitary-pluralist dimensions.	2	1	Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.	1	2	admsci5030148_makarova	1
+Agreed and accounted for.	2	1	I just can't agree that third sector governance is derived from for-profit principles.	1	2	admsci5030148_makarova	1
+Agreed. I misrepresented the analysis. It is not a case study, but instead an cursory archival analysis and exploration, utilizing secondary sources.	2	1	Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.	1	2	admsci5030148_makarova	1
+Agreed. The Analysis, Discussiom, and Conclusion sections are more fleshed out.	2	1	The framework for this section exists but feels a bit superficial and under-developed. I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).	1	2	admsci5030148_makarova	1
+Agreed, and accounted for.	2	1	Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.	1	2	admsci5030148_makarova	1
+I hope that the revisions in the abstract an introduction better clarify the purpose.	2	1	I do not understand the purpose of the paper.	1	2	admsci5030148_makarova	1
+Agreed. I accounted for this throughout the document. Terminology is used in a definitionally consistent manner.	2	1	The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.	1	2	admsci5030148_makarova	1
+Note that I adjusted the paper, and recalibrated it from a case study, to a discussion paper that utilizes an archival analysis of secondary sources.	2	1	The style is more journalistic and free opinion than rigourous research.	1	2	admsci5030148_makarova	1
+Noted and accounted for throughout the document.	2	1	Public enterprises, third sector cooperatives are all mixedup.	1	2	admsci5030148_makarova	1
+Acknowledged. Acknowledged. I cleaned and honed the Introduction, and added a more robust Conclusion.	2	1	The conceptual framework is not clearly presented and I do not see what are the contributions except some free statements at the end not really related to the analysis.	1	2	admsci5030148_makarova	1
+We fundamentally reorganized our manuscript so that its story and structure might be much clearer in the revision. In Section 2.2 we explicitly presented three hypotheses that are tested and discussed in later sections using information on six cluster cases. Section 5 also was reorganized so that the relationship of local cluster management to basic conditions and to national cluster polity, respectively, could be tested for each country using some cases of clusters and cluster management.	2	1	the analysis of the data is weak, and the paper remains largely descriptive. Would suggest to use these case studies as a source of ideas on how policies and cluster initative management migth relate to each other; that would make this a more interesting paper.	1	2	admsci5040213_boyarkin	1
+We revised and simplified Figure 1 by deleting cluster performance and its relationships with other factors because in fact we do not address them in our manuscript.	2	1	The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included.	1	2	admsci5040213_boyarkin	1
+We read and cited in our manuscript the suggested and other references that correspond to [25], [31], [32], [33], [34], [35] and [36] (especially in Section 2.2) in revising our conceptual framework.	2	1	"There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_boyarkin	1
+This manuscript addresses the relationship of national cluster policy and its local implementation, and not the policy organization within the government. Therefore, we slightly changed the titles of the manuscript and Section 5, and revised related terms throughout the manuscript, using the terms such as “local implementation” or “local management”.	2	1	The language the authors use is here a bit imprecise - they seem to be talking about the management of cluster initiatives, not about the management of the policies themselves (which is more a question of how the program oversight is organized within government; also an interesting but different question).	1	2	admsci5040213_boyarkin	1
+The revised version went through a professional, native English editing.	4	1	I would encourage you to have a native speaker look at it, mainly to clarify the language in some places.	3	2	admsci5040213_boyarkin	1
+Indeed, it is not a big surprise that the characteristics of national cluster policies and their local implementation are consistent, but still it is our (small but significant) contribution to concretely show the consistency in international comparison with original interviews of cluster managers. It is beyond the scope of our manuscript to answer the question as to which model is better for the cluster performance, so we leave it as a future research agenda.	4	1	Content-wise to me the key observation is the - potentially systematic - connection between the way cluster policies arestructure and the organization of cluster initiatives. This is not a big suprise, and it doesn't really answer the question asto which model is better (only that there are internally consistent models that are different). But it is still a usual observation to make.	3	2	admsci5040213_boyarkin	1
+We revised the concept and its explanation, and added some references in Section 2.2 and in Introduction. For more details, please see below.	2	1	the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2. It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices. The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters. As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope.	1	2	admsci5040213_boyarkin	1
+In this manuscript, we do not intend to demonstrate direct performance effect of cluster conditions, cluster policy, and cluster management. This is beyond the scope of our paper. It would be an important research topic to challenge the “technocratic faith”, but we cut the discussion about the factors of cluster performance from the text and Figure 1 in order to avoid any misunderstanding. Instead, we added a detailed discussion about “initial conditions” (now “basic conditions”) of clusters in Section 2.2 and 3.1 (a new section).	2	1	the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2. It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices. The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters. As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope.	1	2	admsci5040213_boyarkin	1
+In page 4 below in Section 2.2, we added a brief explanation of the “initial” (now “basic”) conditions of clusters. There we suggested that they comprise various regional characteristics and that also R8 scientific or industrial focus may differ across clusters within life science or biotechnology. In the new Section 3.1, we explained the differences of basic conditions in more detail.	2	1	‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance. Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp. regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.	1	2	admsci5040213_boyarkin	1
+It is true that in some cases local cluster organization had been established and had started its activity before the focal cluster policy started. Because of limited information, we could not sufficiently consider cluster and policy dynamics over time, but referred to the lack of dynamic approach in our study in the conclusion.	2	1	At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.	1	2	admsci5040213_boyarkin	1
+* We recognized that we did not concretely explain the initial (we changed the term to “basic” in the revision) conditions of clusters in the previous version. Moreover, we recognize that it is important to discuss in detail how these countries differ regarding basic conditions and why the national cluster policies differ across them. Therefore, we fundamentally changed the structure of our manuscript and set up a new Section 3.1 to discuss these issues in detail. Here, we cited the suggested book chapter and paper, but mainly used statistical evidence from OECD Science, Technology and Industry Scoreboard on (different) innovation systems to characterize basic conditions of clusters and to distinguish between private- and public-driven clusters. We summarized the discussion in Table 1 (new).	2	1	A conceptual and/or theoretical perspective is needed to shed light at the differences found. in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up. The authors should include a discussion on how these countries differ e.g.	1	2	admsci5040213_boyarkin	1
+Maybe “intellectual cluster” is a specific jargon in the Japanese policy. Following your suggestion, and according to the EU reports we cited in our manuscript, we deleted the adjective “intellectual” for clusters and just use “clusters”, because it does neither affect our concept nor findings.	2	1	The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies. According to the definition on the top of page 4, the term denotes a cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper. Even so, the term ‘intellectual’ does not look fully appropriate here, which may be due to its translation from a Japanese original. ‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters. At the least, it would appear sound to qualify these six cases as science-based clusters, but then this would apply to the biotech industry and all its clusters in general.	1	2	admsci5040213_boyarkin	1
+We did not include USA in the international comparison because, as you correctly suggested, there is no comparable national cluster policy there. However, we use USA in Table 1 on basic conditions as the baseline reference for the three countries.	2	1	The choice of clusters is generally well explained on pp. 2-3. When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!	1	2	admsci5040213_boyarkin	1
+We recognize that the term “representative” is misleading. We replaced it with “outstanding” in the revised manuscript.	2	1	When explaining the choice of regional cases, the term ‘representative’ appears misleading. It seems that the most prominent or successful cases have been selected, at least for Germany and France. How can these be ‘representative’ for whatever population of biotech clusters in these countries?	1	2	admsci5040213_boyarkin	1
+We added detailed information about the number and types of interview partners and interview time on page 9 at the end of Section 2.3 (Research Methodology).	2	1	When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number. Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e. representatives of the respective national cluster programmes, have not been surveyed as well.	1	2	admsci5040213_boyarkin	1
+* We did not interview the officers in charge of cluster policy in the government because information on national cluster policy was sufficiently available from cluster managers and second sources including ministry’s websites and because we are more interested in the relationship of cluster policy with basic conditions and local management than in the cluster policy itself.	2	1	When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number. Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e. representatives of the respective national cluster programmes, have not been surveyed as well.	1	2	admsci5040213_boyarkin	1
+With 3 references, we discussed in the (newly inserted) third paragraph of Section 2.3 the advantages of comparative case studies across countries focusing on biotechnology.	2	1	some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_boyarkin	1
+The classification between top-down and bottom-up clusters is misleading. In the conceptual framework we distinguish between public-driven and private-driven clusters, so that in the revised version we focused on this type of distinction and cut all descriptions on top-down and bottom-up clusters.	2	1	sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between. Compared to the Japanese cases, the French and German cases are clearly more bottom-up, but they still display clear – if not dominant – elements of top-down governance and public agency. This dichotomy should be refined and the classification of cases qualified as, e.g., ‘relatively bottom-up’.	1	2	admsci5040213_boyarkin	1
+Referring to Kiese (2013) and Sternberg et al. (2010), but also other papers such as Crespy et al. (2007) for France, we added that, especially for Germany, we should not underestimate the roles of states and local authorities in the cluster policy. Finally, in the conclusion, we stressed the lack of multilevel governance approach as a limitation, which should be more explicitly considered in future research.	2	1	For the German cases, the authors may have underestimated public agency, since they did not investigate the role of state governments (Bavaria, Baden-Württemberg) within the country’s system of multilevel governance, which is key to understand cluster policies in Germany (cf. KIESE 2013). Furthermore, the state government of Bavaria may be a regional government, but it is certainly not a local authority (like the city of Munich; page 15, line 29) in Germany’s system of multilevel governance.	1	2	admsci5040213_boyarkin	1
+We recognized that this section has several problems, but only this section describes and compares local cluster management based on our own interviews, so that this is the core part of our manuscript. Therefore, we fundamentally changed this section in the following way. First, we have some reference in the revised version, whereas this section is mainly based on our interviews. Second, at the beginning of this section, we explained the relevance of this section in our concept, especially with regard to our (new) hypotheses. Third, more importantly, we reorganized this section to a comparison of cluster management across three countries, focusing on the selection procedure of joint R&D R11 projects (and cutting other information such as monitoring process and support programs). In this way, we made it clear what we do in this section: to check the consistency of local cluster management (regarding project selection) with the cluster’s basic conditions and the national policy. We believe that we could clarify the relevance of the content of this section with the preceding part and the conclusion of this manuscript.	2	1	Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment. This is probably the result of a rather weak empirical basis, provided it draws on interviews with a total of six cluster managers. Critical issues such as the cluster management organisation’s budget and finance have not been addressed. The relevance of the information presented here for the following discussion and conclusion remain far from clear. In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.	1	2	admsci5040213_boyarkin	1
+Cluster Organization’s governance and financing, which was already described in another section in the previous version, is now explained even earlier, in Section 3.2.	2	1	Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment. This is probably the result of a rather weak empirical basis, provided it draws on interviews with a total of six cluster managers. Critical issues such as the cluster management organisation’s budget and finance have not been addressed. The relevance of the information presented here for the following discussion and conclusion remain far from clear. In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.	1	2	admsci5040213_boyarkin	1
+We cut the most part of the discussion in the conclusion (which was mostly redundant with the contents of preceding sections), and added policy implication and some limitations (lack of multilevel governance and dynamic perspectives, possible selection bias of target clusters, and the descriptive and qualitative nature of the study) combined with future research perspectives.	2	1	As also elaborated above, the conclusion should go beyond a mere summary to include methodological reflection, implications for policy (learning), as well as a more extensive discussion of further research perspectives.	1	2	admsci5040213_boyarkin	1
+We cited this paper in footnote 4 in page 3.	2	1	Page 3, line 3: The authors might want to link to the literature on differentiated knowledge bases, which assigns an analytical (science-based) knowledge base to the biotech industry (cf. ASHEIM ET AL. 2011).	1	2	admsci5040213_boyarkin	1
+We agree with the suggestion and changed “few” to “no”. We also checked all the other terms “few” in the manuscript and treated them in the suggested way.	2	1	Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references. If ‘few’ means ‘no’, then please say so straightaway.	1	2	admsci5040213_boyarkin	1
+We changed the sentence (now page 6, line 22) in the following way: “may enhance the incentives of member firms to succeed and partially prevent moral hazard and crowding out”.	2	1	Page 6, line 25: I would suggest softening the statement that matched funding ‘prevents moral hazard and crowding out’. It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.	1	2	admsci5040213_boyarkin	1
+Based on the argument in Section 2.2, we presented three hypotheses in page 7, which are repeatedly addressed in later sections and the conclusion.	2	1	Section 2 contains a number of “expectations”, which could be flagged out and numbered as hypotheses. Taking these up in the empirical discussion could help the reader’s orientation.	1	2	admsci5040213_boyarkin	1
+We found no evidence on policy learning between Germany and France.	2	1	Page 8, lines 1-2: This is interesting indeed. Is there comparable evidence on policy learning between Germany and France? Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.	1	2	admsci5040213_boyarkin	1
+We changed the phrase to “a series of national cluster policy” (now page 18, line 5) at the beginning of Section 4.2.	2	1	Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.	1	2	admsci5040213_boyarkin	1
+We noticed in footnote 18 in page 18 that some cluster regions may have applied for this program twice or three times after failing in the previous competition.	2	1	Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.	1	2	admsci5040213_boyarkin	1
+We cited these studies in Introduction and in Section 4.3 (page 18, footnotes 20 and 21).	2	1	Section 3.3: There is a little bit of literature on French national cluster policy that deserves a brief review or at least reference here	1	2	admsci5040213_boyarkin	1
+We cut the previous Table 2 (on each target cluster and cluster management) in the revision and also the related sentence.	2	1	Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.	1	2	admsci5040213_boyarkin	1
+We agree with you. However, according to the MEXT’s definition, clusters need not involve local high-tech ventures. R&D collaboration between local public research institute and private firms outside of the cluster area is also an important part of a cluster for MEXT. Regarding the aim of local authorities, which includes the development of a cluster around a research organization or hospital, indeed cluster policy has not produced any outcomes yet.	2	1	Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’? If the aim of public policy is to grow a cluster around a research organisation or hospital, this does not seem to produce any meaningful outcomes as yet.	1	2	admsci5040213_boyarkin	1
+We used “prominent” and “excellent” instead of “famous”.	2	1	Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’. This should be backed up with evidence (data) and preferably rephrased, such as ‘recognised research universities’, or ‘elite’ with reference to the German federal government’s programme of university excellence (Exzellenzinitiative).	1	2	admsci5040213_boyarkin	1
+We cut this sentence.	2	1	Page 15, line 27: Repetition – the IZB incubator was already mentioned on line 9.	1	2	admsci5040213_boyarkin	1
+We cited this paper in footnote 14 in Section 3.2. (3) to mention that Trinational BioValley has hardly had any impact in terms of intensifying cross-border interaction.	2	1	Section 4.5: Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that KOSCHATZKY (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.	1	2	admsci5040213_boyarkin	1
+We changed the layout of the tables enlarging the font.	2	1	The layout of the tables may be improved, especially the spacing of the text.	1	2	admsci5040213_boyarkin	1
+In the revision, we write the funding amount for Japan in Yen, but give the converged amount in Euro in parentheses. We use the most recent exchange rate for convenience, and explain it where it is first mentioned (note to Table 1 in page 16 and footnote 17 in page 17).	2	1	The paper contains funding information in Yen and Euros. This should be harmonised, or preferably conversions given in brackets or footnotes for the reader’s convenience.	1	2	admsci5040213_boyarkin	1
+We added a discussion on the national systems of innovation in the first two paragraphs of Section 3.1 (p. 9) referring to the suggested seminal works. Moreover, in the remaining part of this section (pp. 10-11), we added some sentences with supporting references on the underlying institutional differences.	4	1	Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators. If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences. This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation. A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).	3	2	admsci5040213_boyarkin	1
+At the beginning of Section 3.2, as well as at the end of the description of each cluster, we mitigated the dichotomist nature in the typology of cluster development by some qualification. For example, we changed the formulation from “characterized as public-driven clusters” to “characterized as more public-driven than private-driven clusters”.	4	1	Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique. I would still like to see a qualification stating that private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.	3	2	admsci5040213_boyarkin	1
+We added a new reference on the development in Japan (Okubo and Tomiura 2010) but deleted the sentence about France because we could not show a reference in English.	4	1	P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.	3	2	admsci5040213_boyarkin	1
+According to this suggestion, we dropped footnote 14 with the related reference.	4	1	P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.” In response, the authors now cite this paper in footnote 14 in Section 3.2. to support the view that BioValley has hardly had any impact in terms of intensifying cross-border interaction. This reformulation is incorrect as the Koschatzky (2000) paper pre-dated the BioValley initiative and does not look at INTERREG either. Unfortunately, my claim regarding the limited impact of BioValley on cross-border networking draws on conversations with practitioners and a field trip to the region, but cannot be supported by a reference as far as I know. I would therefore suggest to drop this footnote altogether.	3	2	admsci5040213_boyarkin	1
+We added the information sources under the table.	4	1	P. 16: Sources should be stated underneath table 2.	3	2	admsci5040213_boyarkin	1
+We replaced “policy” by “programs” in this sentence.	4	1	P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.	3	2	admsci5040213_boyarkin	1
+In Conclusion, we presented again three hypotheses in a new paragraph in Conclusion after the first paragraph (p. 22). Then, in the following paragraph (p. 23) we added a discussion on how differences in cluster policies and cluster management are linked to those in national systems of innovation, with three sentences.	4	1	P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?	3	2	admsci5040213_boyarkin	1
+We fundamentally reorganized our manuscript so that its story and structure might be much clearer in the revision. In Section 2.2 we explicitly presented three hypotheses that are tested and discussed in later sections using information on six cluster cases. Section 5 also was reorganized so that the relationship of local cluster management to basic conditions and to national cluster polity, respectively, could be tested for each country using some cases of clusters and cluster management.	2	1	But the analysis of the data is weak, and the paper remains largely descriptive. Would suggest to use these case studies as a source of ideas on how policies and cluster initative management migth relate to each other; that would make this a more interesting paper.	1	2	admsci5040213_devetyaro	1
+We revised and simplified Figure 1 by deleting cluster performance and its relationships with other factors because in fact we do not address them in our manuscript.	2	1	"The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included. There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_devetyaro	1
+We read and cited in our manuscript the suggested and other references that correspond to [25], [31], [32], [33], [34], [35] and [36] (especially in Section 2.2) in revising our conceptual framework.	2	1	"The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included. There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_devetyaro	1
+This manuscript addresses the relationship of national cluster policy and its local implementation, and not the policy organization within the government. Therefore, we slightly changed the titles of the manuscript and Section 5, and revised related terms throughout the manuscript, using the terms such as “local implementation” or “local management”.	2	1	The language the authors use is here a bit imprecise - they seem to be talking about the management of cluster initiatives, not about the management of the policies themselves (which is more a question of how the program oversight is organized within government; also an interesting but different question).	1	2	admsci5040213_devetyaro	1
+Indeed, it is not a big surprise that the characteristics of national cluster policies and their local implementation are consistent, but still it is our (small but significant) contribution to concretely show the consistency in international comparison with original interviews of cluster managers. It is beyond the scope of our manuscript to answer the question as to which model is better for the cluster performance, so we leave it as a future research agenda.	4	1	Content-wise to me the key observation is the - potentially systematic - connection between the way cluster policies arestructure and the organization of cluster initiatives. This is not a big suprise, and it doesn't really answer the question asto which model is better (only that there are internally consistent models that are different). But it isstill a usual observation to make.	3	2	admsci5040213_devetyaro	1
+We revised the concept and its explanation, and added some references in Section 2.2 and in Introduction. For more details, please see below.	2	1	However, the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2.	1	2	admsci5040213_devetyaro	1
+In this manuscript, we do not intend to demonstrate direct performance effect of cluster conditions, cluster policy, and cluster management. This is beyond the scope of our paper. It would be an important research topic to challenge the “technocratic faith”, but we cut the discussion about the factors of cluster performance from the text and Figure 1 in order to avoid any misunderstanding. Instead, we added a detailed discussion about “initial conditions” (now “basic conditions”) of clusters in Section 2.2 and 3.1 (a new section).	2	1	As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope. The performance of clusters is also affected by many exogenous influences not captured in the simple model.	1	2	admsci5040213_devetyaro	1
+In page 4 below in Section 2.2, we added a brief explanation of the “initial” (now “basic”) conditions of clusters. There we suggested that they comprise various regional characteristics and that also R8 scientific or industrial focus may differ across clusters within life science or biotechnology.	2	1	Furthermore, ‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance. Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp. regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.	1	2	admsci5040213_devetyaro	1
+It is true that in some cases local cluster organization had been established and had started its activity before the focal cluster policy started. Because of limited information, we could not sufficiently consider cluster and policy dynamics over time, but referred to the lack of dynamic approach in our study in the conclusion.	2	1	At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.	1	2	admsci5040213_devetyaro	1
+We recognized that we did not concretely explain the initial (we changed the term to “basic” in the revision) conditions of clusters in the previous version. Moreover, we recognize that it is important to discuss in detail how these countries differ regarding basic conditions and why the national cluster policies differ across them. Therefore, we fundamentally changed the structure of our manuscript and set up a new Section 3.1 to discuss these issues in detail. Here, we cited the suggested book chapter and paper, but mainly used statistical evidence from OECD Science, Technology and Industry Scoreboard on (different) innovation systems to characterize basic conditions of clusters and to distinguish between private- and public-driven clusters. We summarized the discussion in Table 1 (new).	2	1	However, it remains rather descriptive and does not systematically attempt to explain the differences found between the three countries.	1	2	admsci5040213_devetyaro	1
+Maybe “intellectual cluster” is a specific jargon in the Japanese policy. Following your suggestion, and according to the EU reports we cited in our manuscript, we deleted the adjective “intellectual” for clusters and just use “clusters”, because it does neither affect our concept nor findings.	2	1	According to the definition on the top of page 4, the term denotes a cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper. Even so, the term ‘intellectual’ does not look fully appropriate here, which may be due to its translation from a Japanese original. ‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters. At the least, it would appear sound to qualify these six cases as science-based clusters, but then this would apply to the biotech industry and all its clusters in general.	1	2	admsci5040213_devetyaro	1
+We did not include USA in the international comparison because, as you correctly suggested, there is no comparable national cluster policy there. However, we use USA in Table 1 on basic conditions as the baseline reference for the three countries.	2	1	When explaining the choice of regional cases, the term ‘representative’ appears misleading.	1	2	admsci5040213_devetyaro	1
+We recognize that the term “representative” is misleading. We replaced it with “outstanding” in the revised manuscript.	2	1	When explaining the choice of regional cases, the term ‘representative’ appears misleading.	1	2	admsci5040213_devetyaro	1
+We added detailed information about the number and types of interview partners and interview time on page 9 at the end of Section 2.3 (Research Methodology).	2	1	When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number.	1	2	admsci5040213_devetyaro	1
+We did not interview the officers in charge of cluster policy in the government because information on national cluster policy was sufficiently available from cluster managers and second sources including ministry’s websites and because we are more interested in the relationship of cluster policy with basic conditions and local management than in the cluster policy itself.	2	1	Furthermore, some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_devetyaro	1
+With 3 references, we discussed in the (newly inserted) third paragraph of Section 2.3 the advantages of comparative case studies across countries focusing on biotechnology.	2	1	Furthermore, some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_devetyaro	1
+The classification between top-down and bottom-up clusters is misleading. In the conceptual framework we distinguish between public-driven and private-driven clusters, so that in the revised version we focused on this type of distinction and cut all descriptions on top-down and bottom-up clusters.	2	1	However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between.	1	2	admsci5040213_devetyaro	1
+Referring to Kiese (2013) and Sternberg et al. (2010), but also other papers such as Crespy et al. (2007) for France, we added that, especially for Germany, we should not underestimate the roles of states and local authorities in the cluster policy. Finally, in the conclusion, we stressed the lack of multilevel governance approach as a limitation, which should be more explicitly considered in future research.	2	1	However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between.	1	2	admsci5040213_devetyaro	1
+We recognized that this section has several problems, but only this section describes and compares local cluster management based on our own interviews, so that this is the core part of our manuscript. Therefore, we fundamentally changed this section in the following way. First, we have some reference in the revised version, whereas this section is mainly based on our interviews. Second, at the beginning of this section, we explained the relevance of this section in our concept, especially with regard to our (new) hypotheses. Third, more importantly, we reorganized this section to a comparison of cluster management across three countries, focusing on the selection procedure of joint R&D R11 projects (and cutting other information such as monitoring process and support programs). In this way, we made it clear what we do in this section: to check the consistency of local cluster management (regarding project selection) with the cluster’s basic conditions and the national policy. We believe that we could clarify the relevance of the content of this section with the preceding part and the conclusion of this manuscript.	2	1	Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment.	1	2	admsci5040213_devetyaro	1
+Cluster Organization’s governance and financing, which was already described in another section in the previous version, is now explained even earlier, in Section 3.2.	2	1	Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment.	1	2	admsci5040213_devetyaro	1
+We cut the most part of the discussion in the conclusion (which was mostly redundant with the contents of preceding sections), and added policy implication and some limitations (lack of multilevel governance and dynamic perspectives, possible selection bias of target clusters, and the descriptive and qualitative nature of the study) combined with future research perspectives.	2	1	As also elaborated above, the conclusion should go beyond a mere summary to include methodological reflection, implications for policy (learning), as well as a more extensive discussion of further research perspectives.	1	2	admsci5040213_devetyaro	1
+We cited this paper in footnote 4 in page 3.	2	1	Page 3, line 3: The authors might want to link to the literature on differentiated knowledge bases, which assigns an analytical (science-based) knowledge base to the biotech industry (cf. ASHEIM ET AL. 2011).	1	2	admsci5040213_devetyaro	1
+We agree with the suggestion and changed “few” to “no”. We also checked all the other terms “few” in the manuscript and treated them in the suggested way.	2	1	Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references. If ‘few’ means ‘no’, then please say so straightaway.	1	2	admsci5040213_devetyaro	1
+We changed the sentence (now page 6, line 22) in the following way: “may enhance the incentives of member firms to succeed and partially prevent moral hazard and crowding out”.	2	1	Page 6, line 25: I would suggest softening the statement that matched funding ‘prevents moral hazard and crowding out’. It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.	1	2	admsci5040213_devetyaro	1
+Based on the argument in Section 2.2, we presented three hypotheses in page 7, which are repeatedly addressed in later sections and the conclusion.	2	1	Section 2 contains a number of “expectations”, which could be flagged out and numbered as hypotheses. Taking these up in the empirical discussion could help the reader’s orientation.	1	2	admsci5040213_devetyaro	1
+We found no evidence on policy learning between Germany and France.	2	1	Page 8, lines 1-2: This is interesting indeed. Is there comparable evidence on policy learning between Germany and France? Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.	1	2	admsci5040213_devetyaro	1
+We changed the phrase to “a series of national cluster policy” (now page 18, line 5) at the beginning of Section 4.2.	2	1	Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.	1	2	admsci5040213_devetyaro	1
+We noticed in footnote 18 in page 18 that some cluster regions may have applied for this program twice or three times after failing in the previous competition.	2	1	Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.	1	2	admsci5040213_devetyaro	1
+We cited these studies in Introduction and in Section 4.3 (page 18, footnotes 20 and 21).	2	1	Section 3.3: There is a little bit of literature on French national cluster policy that deserves a brief review or at least reference here (e.g., BRETTE/CHAPPOZ 2007, DURANTON ET AL. 2010, LONGHI/ROCHHIA 2013).	1	2	admsci5040213_devetyaro	1
+We cut the previous Table 2 (on each target cluster and cluster management) in the revision and also the related sentence.	2	1	Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.	1	2	admsci5040213_devetyaro	1
+We agree with you. However, according to the MEXT’s definition, clusters need not involve local high-tech ventures. R&D collaboration between local public research institute and private firms outside of the cluster area is also an important part of a cluster for MEXT. Regarding the aim of local authorities, which includes the development of a cluster around a research organization or hospital, indeed cluster policy has not produced any outcomes yet.	2	1	Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’? If the aim of public policy is to grow a cluster around a research organisation or hospital, this does not seem to produce any meaningful outcomes as yet.	1	2	admsci5040213_devetyaro	1
+We used “prominent” and “excellent” instead of “famous”.	2	1	Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’. This should be backed up with evidence (data) and preferably rephrased, such as ‘recognised research universities’, or ‘elite’ with reference to the German federal government’s programme of university excellence (Exzellenzinitiative).	1	2	admsci5040213_devetyaro	1
+We cut this sentence.	2	1	Page 15, line 27: Repetition – the IZB incubator was already mentioned on line 9.	1	2	admsci5040213_devetyaro	1
+We cited this paper in footnote 14 in Section 3.2. (3) to mention that Trinational BioValley has hardly had any impact in terms of intensifying cross-border interaction.	2	1	Section 4.5: Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that KOSCHATZKY (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.	1	2	admsci5040213_devetyaro	1
+We changed the layout of the tables enlarging the font.	2	1	The layout of the tables may be improved, especially the spacing of the text.	1	2	admsci5040213_devetyaro	1
+In the revision, we write the funding amount for Japan in Yen, but give the converged amount in Euro in parentheses. We use the most recent exchange rate for convenience, and explain it where it is first mentioned (note to Table 1 in page 16 and footnote 17 in page 17).	2	1	The paper contains funding information in Yen and Euros. This should be harmonised, or preferably conversions given in brackets or footnotes for the reader’s convenience.	1	2	admsci5040213_devetyaro	1
+The revised version went through a professional, native English editing. Thank you for your recommendation. We provided minor corrections to our manuscript based on the following comments. The revised version went through a professional, native English editing.	4	1	I wish to congratulate the authors for their careful and thoughtful revision. All remarks have been considered, most have been fully incorporated and the paper has gained substantially from revision. As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing. Since this may be managed by the editorial office, I do not see the need for another review.	3	2	admsci5040213_devetyaro	1
+We added a discussion on the national systems of innovation in the first two paragraphs of Section 3.1 (p. 9) referring to the suggested seminal works. Moreover, in the remaining part of this section (pp. 10-11), we added some sentences with supporting references on the underlying institutional differences.	4	1	Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators. If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences. This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation. A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).	3	2	admsci5040213_devetyaro	1
+At the beginning of Section 3.2, as well as at the end of the description of each cluster, we mitigated the dichotomist nature in the typology of cluster development by some qualification. For example, we changed the formulation from “characterized as public-driven clusters” to “characterized as more public-driven than private-driven clusters”.	4	1	Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique. I would still like to see a qualification stating that private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.	3	2	admsci5040213_devetyaro	1
+We added a new reference on the development in Japan (Okubo and Tomiura 2010) but deleted the sentence about France because we could not show a reference in English.	4	1	P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.	3	2	admsci5040213_devetyaro	1
+According to this suggestion, we dropped footnote 14 with the related reference.	4	1	P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.” In response, the authors now cite this paper in footnote 14 in Section 3.2. to support the view that BioValley has hardly had any impact in terms of intensifying cross-border interaction. This reformulation is incorrect as the Koschatzky (2000) paper pre-dated the BioValley initiative and does not look at INTERREG either. Unfortunately, my claim regarding the limited impact of BioValley on cross-border networking draws on conversations with practitioners and a field trip to the region, but cannot be supported by a reference as far as I know. I would therefore suggest to drop this footnote altogether.	3	2	admsci5040213_devetyaro	1
+We added the information sources under the table.	4	1	P. 16: Sources should be stated underneath table 2.	3	2	admsci5040213_devetyaro	1
+We replaced “policy” by “programs” in this sentence.	4	1	P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.	3	2	admsci5040213_devetyaro	1
+In Conclusion, we presented again three hypotheses in a new paragraph in Conclusion after the first paragraph (p. 22). Then, in the following paragraph (p. 23) we added a discussion on how differences in cluster policies and cluster management are linked to those in national systems of innovation, with three sentences.	4	1	P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?	3	2	admsci5040213_devetyaro	1
+We added Freud’s heritage and contemporary research e.g. Jung’s contribution to the Big 5.	2	1	Why Jung? The authors have selected Jung’s archetypes as the theoretical foundation for the paper.	1	2	admsci6020005_makarova	1
+We took great effort introducing above mentioned perspectives into our paper.	2	1	This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.	1	2	admsci6020005_makarova	1
+We decided to call them both gurus because David obviously is the one but our paper refers to Jennifer.	2	1	The abstract states: “Aaker is seen by many as the branding guru” and refers to Aaker (1997) in the introductory paragraph. Are you referring to David Aaker (the father) or Jennifer Aaker (the daughter)?	1	2	admsci6020005_makarova	1
+We added Allen & Olson and other limitations such as criticism on the conceptual, methodological and substantive level as well as confusing user profiles (e.g., upper class) with brand characteristics and weak discriminatory power.	2	1	Indeed, Aaker (1997) is a highly-cited work. Are there any other limitations or criticisms the reader should be informed of? However, you may want to tell readers how brands can convey aspects of personality via advertising (see: Allen and Olson 1995).	1	2	admsci6020005_makarova	1
+We revised the statement in the sense that it is part and not different from it.	2	1	Page 2, line 62: I disagree with the claim that “above models can be seen as extensions of Aaker’s (1997) brand personality model.” The models you listed are variants of well-known hierarchy of effects models.	1	2	admsci6020005_makarova	1
+We fully agree and deleted the claim to be Grounded Theory.	2	1	Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition.	1	2	admsci6020005_makarova	1
+We cited Roberts (2010) who, based on extensive research, found different leading archetypes depending on the product category. We added our thoughts as scope for further research in chapter 4.We added Matzler et al. (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) as research on customer-brand relationship.	2	1	The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.	1	2	admsci6020005_makarova	1
+The survey was conducted face-to-face in the classroom and students asked personally.	2	1	Evidence of claim? Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way. In contrast the 55 Asian students knew the chains.” Was brand familiarity actually measured, or was it assumed?	1	2	admsci6020005_makarova	1
+We added more screenshots and background on the commercials.	2	1	However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).	1	2	admsci6020005_makarova	1
+We added the questions that participants were asked. Everything was based on pencil and paper.	2	1	Clarity and elaboration of Page 5+: The methods section lacks clarity – it would be very difficult for a researcher to replicate the procedure. Please provide greater detail of the scales used, the anchor terms, and how the survey/rating task was administered (online? Paper and pencil?).	1	2	admsci6020005_makarova	1
+Changed in the term ‘manifestation.’ Findings: Who rated the three personality traits?	2	1	“Archetype Articulate Charisma” (heading title in Table 3): This is not clear. Either define the term in the text or use phrasing that is clear to the reader.	1	2	admsci6020005_makarova	1
+The students.	2	1	Findings: Who rated the three personality traits? The student/respondents? How many items were used? Why do you not report reliability measures for the items and scale?	1	2	admsci6020005_makarova	1
+Principal Component with Varimax rotation.	2	1	Factor analysis: Have you reported principal components, or a rotated solution?	1	2	admsci6020005_makarova	1
+We changed the wording. In fact, the cited Cinderalla is a prime example of blended archetype.	2	1	Interpretation: The interpretation of Archetypes seems to assume that the types are mutually-exclusive. However, this is not really the case.	1	2	admsci6020005_makarova	1
+We cleaned up our description of the experimental design by adding the questions that we asked, the scale we used etc.	2	1	I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some.	1	2	admsci6020005_makarova	1
+We tried our best to address this problem, see the revised version of our paper.	2	1	I think that the relevance of the research to practitioners as well as academics needs to include some current findings about ads, their placement and effectiveness.	1	2	admsci6020005_makarova	1
+We added Freud’s heritage and contemporary research e.g. Jung’s contribution to the Big 5. We took the liberty to refer to Kant and Goethe re the origin of the term ‘archetype’ which may give it a more philosophical touch. We would have loved to substantiate the archetype concept by neurophysiological research but were not convinced about the reliability of claimed research findings so we added it under scope for further research (chapter: Implications).	2	1	Why Jung? The authors have selected Jung’s archetypes as the theoretical foundation for the paper. While Jungian approaches have been used by marketing and consumer researchers in the past (e.g., Hirschman), the reasons for this choice need to be justified in the context of other alternatives. Jungian approaches suffer from two core limitations: (a) the assertion of universality – which the authors note, and (b) the psychoanalytic heritage of the ideas (which presupposes a certain view of motivation) is disavowed by most contemporary psychologists.	1	2	admsci6020005_perova	1
+We took great effort introducing above mentioned perspectives into our paper. It stretches across the whole paper. We also mentioned the difference between semiotic and formalist approach.	2	1	Literary archetypes: An alternative approach to studying the archetypal aspects of brand image can be found by scholars who adopt a literary or cultural view of archetypes, such as the ones advanced by Joseph Campbell or Northrop Frye (see, for example: Stern 1989). One could make the argument that using archetypes in advertising has greater affinities to mythology, literature and communications. Work on narrative theory and characterization in advertising also aligns with the archetype approach (see, for example: Mulvey and Medina 2003; Padgett and Allen 1997; Scott 1994; Stern 1988). This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.	1	2	admsci6020005_perova	1
+We decided to call them both gurus because David obviously is the one but our paper refers to Jennifer.	2	1	The abstract states: “Aaker is seen by many as the branding guru” and refers to Aaker (1997) in the introductory paragraph. Are you referring to David Aaker (the father) or Jennifer Aaker (the daughter)?	1	2	admsci6020005_perova	1
+We added Allen & Olson and other limitations such as criticism on the conceptual, methodological and substantive level as well as confusing user profiles (e.g., upper class) with brand characteristics and weak discriminatory power.	2	1	Indeed, Aaker (1997) is a highly-cited work. Are there any other limitations or criticisms the reader should be informed of? There has been some backlash to the application of brand personality to inanimate objects. This may be of concern, because the paper uses hotels as an example. Personally, I agree with you – brand personality is a key facet of brand identity. However, you may want to tell readers how brands can convey aspects of personality via advertising (see: Allen and Olson 1995). This is highly relevant to answering the “so what?” question that is addressed in the implications section.	1	2	admsci6020005_perova	1
+We revised the statement in the sense that it is part and not different from it.	2	1	Page 2, line 62: I disagree with the claim that “above models can be seen as extensions of Aaker’s (1997) brand personality model.” The models you listed are variants of well-known hierarchy of effects models. Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).	1	2	admsci6020005_perova	1
+We fully agree and deleted the claim to be Grounded Theory.	2	1	Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition. Grounded theory is based on the idea of immersion in data to induct new conceptual categories. I see the present study as using existing typologies and categories – there are no new concepts that emerge from the research that can be applied directly to future research.	1	2	admsci6020005_perova	1
+We cited Roberts (2010) who, based on extensive research, found different leading archetypes depending on the product category. We added our thoughts as scope for further research in chapter 4.We added Matzler et al. (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) as research on customer-brand relationship.	2	1	Sample size: A critical view would state that the present research relies on a very small sample of advertisements (n=2) to make its claims. After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity. The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression. Though such studies are rare, good interpretive and quantitative examples can be found in the literature. At a minimum, this exploratory paper should set a more specific and ambitious plan for systematic future research on the topic. For example, Padgett and Mulvey (2007) illustrate a method to identify personal values conveyed by 16 ad campaigns (another complementary dimension of personified brands) and Padgett and Mulvey (2009) demonstrate a novel way to characterize customer-brand relationship archetypes. Imagine a conversation with these authors: what do you contribute to the ongoing conversation?	1	2	admsci6020005_perova	1
+The survey was conducted face-to-face in the classroom and students asked personally. The two Asian hotel chains are globally not very prominent. Most Western executives stay at well-known global hotel chains (e.g., Accor, Hilton) because they have a corporate contract. Shangri-La and Banyan Tree are not that visible in Western countries. This stays in contrast to Asian executives.	2	1	Evidence of claim? Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way. In contrast the 55 Asian students knew the chains.” Was brand familiarity actually measured, or was it assumed? The claim is very absolute – I wonder if any of them ever travelled internationally – presumably EMBAs would be more worldly or cosmopolitan than this… (d)	1	2	admsci6020005_perova	1
+We added more screenshots and background on the commercials. Whereas the BT commercials features beautiful pictures that are sequence-wise interchangeable, the BT commercial unfolds and has it climax at the end with the wolves warming the traveler.	2	1	Figure 3: Because you are studying commercials (which unfold over time), I don’t think the single screenshots really add much value to the paper. However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).	1	2	admsci6020005_perova	1
+We added the questions that participants were asked. Everything was based on pencil and paper.	2	1	Clarity and elaboration of Page 5+: The methods section lacks clarity – it would be very difficult for a researcher to replicate the procedure. Please provide greater detail of the scales used, the anchor terms, and how the survey/rating task was administered (online? Paper and pencil?).	1	2	admsci6020005_perova	1
+Changed in the term ‘manifestation.’ Findings: Who rated the three personality traits?	2	1	“Archetype Articulate Charisma” (heading title in Table 3): This is not clear. Either define the term in the text or use phrasing that is clear to the reader. Please don’t expect them to consult original sources by Jung!	1	2	admsci6020005_perova	1
+The students. We added that part.	2	1	Findings: Who rated the three personality traits? The student/respondents? How many items were used? Why do you not report reliability measures for the items and scale?	1	2	admsci6020005_perova	1
+Principal Component with Varimax rotation. We added in the text.	2	1	Factor analysis: Have you reported principal components, or a rotated solution? More details are required.	1	2	admsci6020005_perova	1
+We changed the wording. In fact, the cited Cinderalla is a prime example of blended archetype.	2	1	Interpretation: The interpretation of Archetypes seems to assume that the types are mutually-exclusive. However, this is not really the case. A close reading of some of the cited work (i.e., Mark and Pearson 1991) recognizes that archetypes can be blended.	1	2	admsci6020005_perova	1
+We hope that we have given full consideration to all comments. We found them very useful and they increased the value of our paper considerably.	2	1	I hope my suggestions are given full consideration, as I believe that they will assist them in achieving this important objective.	1	2	admsci6020005_perova	1
+We cleaned up our description of the experimental design by adding the questions that we asked, the scale we used etc. We included the example of David Beckham. In this context we cited Matzler et al (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) who looked into the archetype-brand-consumer match. We are delighted that our reviewer sees the link (as the authors do) between Aaker and Jung’s images of Freedom, Social, Order and Ego.	2	1	I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some. Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes. It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego. There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.	1	2	admsci6020005_perova	1
+We tried our best to address this problem, see the revised version of our paper. However, this is an uphill task. It goes in the direction of advertising effectiveness. We would have loved to cite some reliable sources on advertising effectiveness e.g. by means of neurophysiological evidence based research but tried in vain.	2	1	I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some. Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes. It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego. There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.	1	2	admsci6020005_perova	1
+line 76 and 79: Sentence about two farms with financial support is deleted from the aim.	2	1	In the introduction, (Line 76 79), you present the objectives, but those are not clear, I respectfully suggest focusing on your hypothesis 2.	1	2	agriculture12060868_makarova	1
+"Line 78: ""designed to meet behavioural needs...."" is deleted Overall, the objective now is focused on the hypotheses."	2	1	Line 78: You wrote, “designed to meet the basic behavioral needs of the sow at farrowing”, but this is not tested on the paper.	1	2	agriculture12060868_makarova	1
+l 80-83 is now deleted from the objectives 4.	2	1	Lines 80 – 83 moves to discussion section 4.	1	2	agriculture12060868_makarova	1
+Figure 1 must was lost from the last document, and we are so sorry for this. The figure is now included in the new document.	2	1	Figure 1 is missing 5.	1	2	agriculture12060868_makarova	1
+More explanation is included in the figure text of figure 2	2	1	Figure 2 needs more description, maybe include which wall… Please describe units of the dimensions 6.	1	2	agriculture12060868_makarova	1
+Corrections are made in the text of figure 4.	2	1	Figure 4. Please indicate which is “sowconf” and which is “trad”. In the left graph the units are in m2, but in right side it does not have unit, seems ar?	1	2	agriculture12060868_makarova	1
+l 206, l225 Info is changed according to the reviewers request.	2	1	Line 206: I don’t understand why you use just 10 o 12 sows, 50% primi and multiparous, but in line 225 mentioned that “there were no secure information about sow parity” please be consistent.	1	2	agriculture12060868_makarova	1
+Insted of inserting P-values in the text when reporting significant differences for the ones that are in tables, we have included more references to the stat-tables.	2	1	Please include P values when report significant differences.	1	2	agriculture12060868_makarova	1
+This is now included.	2	1	Figure 1 does not appear in the pdf that this reviewer could download. Only the Figure Caption appears.	1	2	agriculture12060868_makarova	1
+Stillborn piglets vs piglets with no milk: more details are included about this in the description of the M/M	2	1	It is not totally clear to the reviewer how stillborn piglets were evaluated as compared to “piglets with no milk in stomach”.	1	2	agriculture12060868_makarova	1
+224-225: confusion about parity is corrected,	2	1	Line 224-225, it states that that no information/security on the productivity data on whether it was primiparous or multiparous sow. Again this creates a bit of confusion. Sows were recorded, 50% primiparous and 50% multiparous to evaluate behaviour, and results are discussed accorging to that (lines 320-321). Why productivity data was not available?	1	2	agriculture12060868_makarova	1
+Figure 5 and 7. There is always effects of batches in these types of studies. This is also why we prefer to have several batches, but the main effect should not be biased by the batch effect in the present study, as the main finding are quite clear even in our small data set.	2	1	Figure 5 and 7 display the overall causes of mortality for the three batches. However, differences in batches were found. Could this overall causes be biased by this batch effect?	1	2	agriculture12060868_makarova	1
+mean + se is given in all tables and figures, Replicates are now included and abbreviations explained.	2	1	Please add the important information to the footnote or figure legend of the tables and figures. Such as: abbreviations, replicates n=?, mean±SD/SE, ect.	1	2	agriculture12060868_makarova	1
+Italic for P-values in the text. Done!	2	1	Please italic the P value throughout the paper.	1	2	agriculture12060868_makarova	1
+Figure 1 was no included by an error in the last MS. It is now inserted again.	2	1	Lines 129-133, where is the figure 1?	1	2	agriculture12060868_makarova	1
+Batches and number of litters are added to the figure legends.	2	1	Line 263, please add “(3 batches with TRAD and 2 batches with SOWCOMF)” to the figure legends.	1	2	agriculture12060868_makarova	1
+Space before and after sign is added.	2	1	Line 275-276, Space needed before and after “±”.	1	2	agriculture12060868_makarova	1
+A new and improved version of figure 6 is now included.	2	1	The quality of the Figrue 6 need to be improved.	1	2	agriculture12060868_makarova	1
+We have inserted * to denote differences instead of letters.	2	1	Figure 7, using “*” to substitute the different letters to express the difference.	1	2	agriculture12060868_makarova	1
+Table 4 is inserted again in the writing form.	2	1	Table 4, please using the writing way to present the table.	1	2	agriculture12060868_makarova	1
+The meaning of ***: this is explained.	2	1	Figure8, do not understand the meaning of “***” here?	1	2	agriculture12060868_makarova	1
+Quality of figure 7 is improved.	2	1	Figure 7, please improve the perspective of the figure.	1	2	agriculture12060868_makarova	1
+We better clarified this in the method section (L101, L105-106). 2000 is the year of the establishment of Conservation and Safety Network. In this frame, seeds were collected and maintained in long term storage conditions by Arsial in the subsequent years.	2	1	lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?	1	2	agronomy12061433_makarova	1
+Between 8-10 grams (about 2000 seeds) we don’t think is a detail to insert in the manuscript.	2	1	How many seeds were collected?	1	2	agronomy12061433_makarova	1
+Yes, we clarified (L116).	2	1	line 115: Did one repetition mean one plant?	1	2	agronomy12061433_makarova	1
+We added these details (L132 and L136) The results are described and discussed in detail.	2	1	line 130: How many fruits were used as a sample of representative fruits?	1	2	agronomy12061433_makarova	1
+We prefer to keep the introduction in order to give a broad overview to readers	2	1	The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.	1	2	agronomy12061433_makarova	1
+"The cluster analysis is based on phenotypic traits, this analysis revealed specific clusters of ""Cornetto di Pontecorvo"" and similar types, although they were not clearly separated at the hierarchical level. The genotyping analysis provided more in-depth insights. We addressed this in the discussions section (see L485 and subsequent)."	2	1	In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green? Any explanation?	1	2	agronomy12061433_makarova	1
+Product is correct since refer to This crop (singular), we specify (as a food and non-food product)	2	1	“products”	1	2	agronomy12061433_makarova	1
+We agree and better positioned the table 3 after the sentence Line 385-388: the figure legend does not provide enough information on this figure #Answer:	2	1	Line 245: would be better to show the value of each agronomic trait mentioned here	1	2	agronomy12061433_makarova	1
+Figure 4 caption has been improved	2	1	Line 385-388: the figure legend does not provide enough information on this figure	1	2	agronomy12061433_makarova	1
+We agree and modified the sentence (see L439)	2	1	Line 435: it may not be worth to mention the sweet pepper as the first 30 crops Line 468: “GBS”?	1	2	agronomy12061433_makarova	1
+We change referring to ddRAD seq (see L473) Conclusion:	2	1	Line 468: “GBS”? Genotype by sequencing?	1	2	agronomy12061433_makarova	1
+In order to answer to this question, we improved the conclusion (L524-529)	2	1	What kind of broader impact of this work could be?	1	2	agronomy12061433_makarova	1
+We better clarified this in the method section (L101, L105-106). 2000 is the year of the establishment of Conservation and Safety Network. In this frame, seeds were collected and maintained in long term storage conditions by Arsial in the subsequent years.	2	1	lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?	1	2	agronomy12061433_perova	1
+Between 8-10 grams (about 2000 seeds) we don’t think is a detail to insert in the manuscript.	2	1	How many seeds were collected?	1	2	agronomy12061433_perova	1
+Yes, we clarified (L116).	2	1	line 115: Did one repetition mean one plant?	1	2	agronomy12061433_perova	1
+We added these details (L132 and L136)	2	1	line 135: How many fruits were used as a bulk of representative fruits?	1	2	agronomy12061433_perova	1
+We have carefully considered comments and thoughtful suggestions, revising the manuscript accordingly. All amendments suggested have been included. Additional corrections through the text have been done.	2	1	Conclusion: What kind of broader impact of this work could be?	1	2	agronomy12061433_perova	1
+We prefer to keep the introduction in order to give a broad overview to readers	2	1	The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.	1	2	agronomy12061433_perova	1
+"The cluster analysis is based on phenotypic traits, this analysis revealed specific clusters of ""Cornetto di Pontecorvo"" and similar types, although they were not clearly separated at the hierarchical level. The genotyping analysis provided more in-depth insights. We addressed this in the discussions section (see L485 and subsequent)."	2	1	In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green? Any explanation?	1	2	agronomy12061433_perova	1
+Product is correct since refer to This crop (singular), we specify (as a food and non-food product)	2	1	Line 37: “products”	1	2	agronomy12061433_perova	1
+We agree and better positioned the table 3 after the sentence	2	1	Line 245: would be better to show the value of each agronomic trait mentioned here	1	2	agronomy12061433_perova	1
+Figure 4 caption has been improved	2	1	Line 385-388: the figure legend does not provide enough information on this figure	1	2	agronomy12061433_perova	1
+We agree and modified the sentence (see L439)	2	1	Line 435: it may not be worth to mention the sweet pepper as the first 30 crops	1	2	agronomy12061433_perova	1
+We change referring to ddRAD seq (see L473)	2	1	Line 468: “GBS”? Genotype by sequencing?	1	2	agronomy12061433_perova	1
+In order to answer to this question, we improved the conclusion (L524-529)	2	1	Conclusion: What kind of broader impact of this work could be?	1	2	agronomy12061433_perova	1
+The content of the two papers Reviewer 1 mentions here focussed on the impact of the pandemic on management changes in dogs and on dog owner experiences at a single time point during the most restrictive ‘lockdown’. The current paper is different to either of these, and indeed any other published paper, in that it focusses on changes in separation related behaviour over time, across two different surveys in a longitudinal cohort. The paper is about separation-related behaviour, not about the pandemic. The changes in leaving routine that occurred during the pandemic afforded the opportunity to study what impact this had on a large population of pet dogs over time, that would otherwise have been impossible to capture at such scale. The topic is unique and important in that it adds evidence for an association that has previously been supposed or observed by individual practitioners, but not demonstrated in the published literature, and certainly not on this scale.	2	1	A catchy topic, but its content is unoriginal, bringing nothing new to the pre-pandemic science. It seems that the authors of this study have already tackled this topic exhaustively in their previous articles	1	2	ani12040482_perova	1
+Yes, this was purposeful, it provides a summary of the aims & methods to introduce the discussion as not everyone reads papers in a linear fashion. We have not made any changes based on this statement.	2	1	The entire first paragraph of Discussion repeats what has already been written in earlier chapters (line 372-379).	1	2	ani12040482_perova	1
+There is a difference between something that is considered to be “well-known” or folk-knowledge and demonstrable evidence. The sentence quoted here is not untrue, or exaggerated, however, if the Editor wishes us to explore this further, we would kindly request the Reviewer provide us with examples of publications where this has been empirically demonstrated, as we have not found any in our own searches.	2	1	"Line 382-384 „To our knowledge, this is the first empirical evidence to show a link between changes in dogs’ leaving routines and risk of SRBs.” The authors take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed. And changing the routine of the day is a well-known factor influencing the dog's welfare and behavior, including the occurrence of separation anxiety, so the above-quoted sentence should be deleted because it is untrue. The authors have hardly studied all the publications on how dogs behave when daily routines were changed, so the term ""first study"" is exaggerated."	1	2	ani12040482_perova	1
+Given the lack of constructive feedback in these statements, no changes have been made based on these comments.	2	1	Line 444 „This study has provided a unique and valuable insight into…..” the authors also mark their only unique role in the study, which does not bring new content.	1	2	ani12040482_perova	1
+Thank you, in line 16 and 32 we have added “when restrictions had eased” to explain why we surveyed in October 2020.	2	1	My only suggestion is that making it clear why February and October were chosen as survey-points may be beneficial. This is discussed later on but highlighting here would be useful.	1	2	ani12040482_perova	1
+Thank you, we have now expanded this to read: “In addition to being a welfare concern for the dog, SRBs are often unwanted or problematic for the owner and/or neighbours such as in the case of excessive vocalisation, which may lead to noise complaints, or in terms of house soiling and destruction of property, which may be particularly problematic for people living in rental accommodation. In such cases, these issues may increase risk of relinquishment [7]” (see new lines 56-59) It may be helpful to discuss why there is poor owner adherence to programmes to reduce SRB (Line 66-67).	2	1	Aa little more mention of issues of SRBs from owner perspective (e.g. noise complaints and issues with housing; property damage; decline of human-pet bond) could be beneficial (line 56-57).	1	2	ani12040482_perova	1
+The following text has now been added beginning at line 69: “Whilst the precise reasons for poor adherence to advice for preventing or reducing SRBs have not been documented, it is at least partly attributed to the relative difficulty of adhering to specific aspects of these programmes, with the easier pieces of advice followed more often than the parts that take more commitment and consideration [13].” I wasn't sure if line 79-80 was needed- about dogs having returned to pre-pandemic pattern, I think it is a little dismissive of your own research which is valuable and applicable beyond the pandemic.	2	1	It may be helpful to discuss why there is poor owner adherence to programmes to reduce SRB (Line 66-67).	1	2	ani12040482_perova	1
+Thank you for your comment, I think we were pre-judging that people would think it was just another covid paper, and we are glad to see you agree with us that it is valuable beyond the pandemic. This line has now been deleted.	2	1	I wasn't sure if line 79-80 was needed- about dogs having returned to pre-pandemic pattern, I think it is a little dismissive of your own research which is valuable and applicable beyond the pandemic.	1	2	ani12040482_perova	1
+The findings were all extremely similar, so it felt quite repetitive to discuss them all. We actually had more detail in an early draft then trimmed it out. However, we have now added some more of this detail as requested (see lines 100 to 112), whilst trying to limit repetition.	2	1	Line 96-97- some more detail on the findings of these various surveys would be beneficial similarly to Bowen's study, just so provide a clear background/discussion of literature setting scene for your study.	1	2	ani12040482_perova	1
+We presumed that at some point owners would pop out of the house, to take out the bin or go to the garage etc.. Such leaving events are likely to be experienced differently by some dogs, as the owners won’t likely give off the same cues as when they leave for an extended period of time, such as picking up house keys, or leaving through a different door. This has been explained now in lines 179-83.	2	1	It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g. justification for this/why this time duration used.	1	2	ani12040482_perova	1
+Thank you, this seemed ok when we submitted it, but must have gone wrong when uploaded but we’ve corrected it now.	2	1	Line 131: There was an error with referencing wanted to flag.	1	2	ani12040482_perova	1
+We’ve amended description of the study periods in a few places, including between lines 132-142, which we hope addresses this query now.	2	1	It may be useful to spell out a little more here why these survey periods were chosen here, this is covered elsewhere but think could be made clearer here and earlier.	1	2	ani12040482_perova	1
+One of the other reviewers also commented on this and asked for Figure 1 and much of the text to be moved to the supplemental material, so It’s also further explained in the supplementary material how it links to working and in lines 140-142 in the main text which now explains that many businesses were open again in October.	2	1	Figure 1- Perhaps just double-check this is referred to in the text. Think either way this figure would benefit a bit of context on why was included/what was showing in context of this study as currently info is a bit generic and didn't totally tie into people's working from home etc.	1	2	ani12040482_perova	1
+This was due to the errors you mentioned below. We used the Word automatic referencing and somehow that seemed to get broken when the document was uploaded to the online system. It’s been done manually now.	2	1	However, I don't think the figures were referred to in the text (unless that was the errors highlighted below?)	1	2	ani12040482_perova	1
+See above.	2	1	There were some errors where reference source not found which would want to look into (line 246; 265; 274; 292; 305; 323 etc).	1	2	ani12040482_perova	1
+Agreed, this was how we interpreted why the owner was reporting this change. We have rephrased this section now.	2	1	Line 366-369= I'm not sure it's totally fair to interpret this (the dog not always bringing toy when greeting them) as an anthropomorphic interpretation on the part of the owner- the anthropomorphic interpretation seems to be from authors of manuscript not owner? They do not seem to state dog is favouring them less unless text was omitted here.	1	2	ani12040482_perova	1
+Thank you for your review. We have expanded the discussion to address these points, however due to another reviewer saying the paper was already too long, we have kept this brief. A section detailing relevance to practitioners has been added to lines 545-551.	2	1	I felt the discussion about changes in greeting behaviour as an early indicator was interesting and could be expanded. Some more detail on the application of the findings to practitioners/owners would be beneficial. I do think this is of interest to a range of stakeholders.	1	2	ani12040482_perova	1
+Yes, this is correct. This is stated in line 232-233 or the original submission, but it seems we omitted it from the Methods section. Thank you for spotting that. Text has been added to lines 198-200 to explain this.	2	1	P6 L176 I am assuming you excluded dogs that were not left alone in February (or were there none?)	1	2	ani12040482_perova	1
+Within the results table (Table 2) the direction of the estimate for this result is negative. Whilst we agree that it would be more impactful worded the other way around, we worry that changing the way it is described won’t match up to the data as it was coded and reported. We haven’t changed this wording for these reasons.	2	1	P10 L307-308 Could you make this a positive statement to show highlight the impact of amount of change on SRB: Dogs whose days left alone per week increased the most…If the data support that angle, I think it’s a stronger statement than what is written.	1	2	ani12040482_perova	1
+Yes, and no, depending on your perspective. Whilst this was a significant predictor in the modelling, in reality 55% of dogs that were being left again changed from SRB+ to SRB-. So whilst 45% of dogs still showed it, a large proportion did not.	2	1	1. That dogs showing SRB prior to lockdown were more likely to show it after. Suggests the behavior is fairly robust and dogs having shown it at one point continue to be at risk (without knowing what interventions were taken)	1	2	ani12040482_perova	1
+Thank you for these thoughtful comments, we agree that we were a bit overly focussed on the results that supported the studies hypothesis and neglected to discuss the findings related to the dogs whose SRB appeared to have resolved. The number and percentage are detailed in the results section (lines 282-283 of the original submission and 298-298 in this one), but we failed to discuss these in the Discussion. We ran two models, one focussing on the dogs who started SRB- to see how many changed to SRB+ or remained SRB- and what factors influenced this, and the other focussing on dogs that started SRB+ to see what factors influence whether they changed to SRB- or remained SRB+. These models therefore, cover the comparisons you suggest (unless we’ve misinterpreted your comments). We have now added some discussion of the dogs who began the pandemic SRB+ however, as this was lacking from the first version (see the paragraph beginning at line 524).	2	1	2. Can you address the dogs that improved between Feb and Oct? You report the number of new dogs showing SRB but can you report on dogs that went the other way? What was the actual # and %? It seems you have compared within dogs that showed SRB in baseline and other models for dogs that did not. Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing. This comparison seems essential—if some dogs get worse but some get better, than the argument that changes due to COVID might bring about new SRB becomes weaker.	1	2	ani12040482_perova	1
+Thank you for this comment, we have discussed this further in the Discussion now (see lines 517-523)	2	1	3.60 One of the challenges in this is we don’t know background rates of change for dogs with SRB. That is, would the same % of dogs changed (in either direction) regardless of changes in owner schedules? We might see some dogs become sensitized over time and develop SRB (or something else changes in their life to precipitate it), while others might habituate or their behavior undergo extinction. As such, we can’t say for certain that these changes are due to changes in owner schedule. You note that the pseudo-R2 was small but I think you could go into more details and note the possibility that it was unrelated too. However, your results that the greater the change → the greater the new risk of SRB does lend support to the idea that this change in management might be a factor. I would suggest highlighting and discussing this outcome more.	1	2	ani12040482_perova	1
+Thank you, we have changed the title to: Impact of changes in time left alone on separation-related behaviour in UK pet dogs	2	1	Title: change “Leaving Pattern” to “Time Spent Away”; leaving pattern is less clear (it could be frequency of leaving each day)	1	2	ani12040482_perova	1
+This is how it was phrased in the survey, so this has not been changed.	2	1	Line 214: if possible (might not be possible if this was how it was phrased in the survey), change “different to before” to “different from before”	1	2	ani12040482_perova	1
+Thank you, this edit has been applied.	2	1	Lines 383-384: Change “dogs’ leaving routines” to “dogs’ time left alone”	1	2	ani12040482_perova	1
+It seemed unfeasible to expect a dog to never be left for even a minute or two, for example if the owner pops outside to take out the bin or to get something from the garage/shed etc.	2	1	Line 151: how was the criterion of “at least 5 minutes” chosen?	1	2	ani12040482_perova	1
+The wording given to the owners is all presented here, and in the supplementary material. No further definitions for behaviours or vocalisations were given. Pined, whined and cried were all mentioned, as all are words owner tend to use to describe similar dog vocalisations. As with all owner awareness of SRBs, it is fundamentally challenging unless they use recording equipment to view their dogs when they leave them or are otherwise able to view their dogs when alone such as by looking through a window. As this is a limitation of the method used, the appropriate place to mention this would be in the Discussion, not the Methods, so we have not discussed this limitation there.	2	1	My main question when reading this section concerned how owners would know that certain separation-related behaviors occurred when they were away? There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing? This topic is not addressed until the Discussion section (line 395) but should be addressed here in the Methods and Materials section. Were behaviors, including different vocalizations, defined for owners? For example, what is the difference between pined, whined, and cried?	1	2	ani12040482_perova	1
+The wording of the question asked about the dog being left alone, without human company (see line 173), so it applied to anyone, and specifically meant when no-human was present in the house with the dog.	2	1	Also, I might have missed it, but did all of these owners live alone with their dog(s), meaning no other people in the household? If other people were present, did all of them have to be away or just the dog’s owner? This information might be presented in your other paper, but it would be good to include this basic information here as well, so readers can better understand your methods and findings.	1	2	ani12040482_perova	1
+This has now been explained, however this section has been moved entirely to the supplementary material on request of other reviewers.	2	1	Line 166: define “devolved nations” for readers unfamiliar with the term.	1	2	ani12040482_perova	1
+Not strictly speaking, no. Dogs who were not left in February were excluded from the analysis. So, whilst there are 0 of them, that’s because of methodological choice not because there weren’t any dogs left alone. A note explaining this has been added to the figure legend.	2	1	Figure 2: should the blank box for “Feb, Not at all” be 0.0%?	1	2	ani12040482_perova	1
+There were issues in the figure referencing when we uploaded the manuscript to the submission system. This has now been corrected.	2	1	Figure 4 is not called out in the text of the Results section.	1	2	ani12040482_perova	1
+For some reason I was convinced I’d read it was optional for longer discussions, but having double checked, you are indeed correct. Thank you. A brief conclusion section has now been added.	2	1	I believe Animals requires a Conclusions section.	1	2	ani12040482_perova	1
+Thank you, we have corrected the typo.	2	1	Line 100: insert “the” after “in”	1	2	ani12040482_perova	1
+This has been corrected	2	1	Line 355: delete the first “the”	1	2	ani12040482_perova	1
+Thank you for your comments. The detail was provided to anchor the study in the background of what was occurring at the time, but we agree it is not necessary to the reader to see in the main text. These details and Figure 1 have been moved to the Supplementary material.	2	1	However, currently, the paper seems too long, and it goes into too much detail about how the pandemic took place in the UK. This information would have been interesting if data on the mood, stress level, general mental health of the owners had also been collected in connection with the pandemic and the links with dog behaviour have been investigated. But the questionnaire only asked how long the dogs had been alone and what they were doing during this time. Therefore, the detailed description of the pandemic is unnecessary and could be moved to the Supplemental material, together with Figure 1.	1	2	ani12040482_perova	1
+We agree this was completely missed from the Discussion. It wasn’t intentional, we were just overly focussed on the results that related to the research hypothesis. Discussion around this has been added now to lines 524-544.	2	1	I think that according to the statistics, the strongest finding is that dogs who showed SRB in February or May had increased odds of having SRB during the subsequent measures which is not surprising. What is surprising though is that 55.7% of the dogs who were reported to show SRB in February (before the pandemic) were clear in October which is good news. However, the authors do not focus on this result and do not explain this finding.	1	2	ani12040482_perova	1
+Agreed, we have altered this part of the simple summary and abstract and included discussion of this in lines 539-544 now as well as into the conclusions, thank you.	2	1	L18-20 „Whether dogs showed SRBs or not changed considerably over the months of the study, and one in ten dogs were found to have developed new SRBs in October, that they didn’t show before the pandemic” Overall, the percentage of dogs showing SRBs decreased during the study, and this should be clarified in the sentence above. What I see in Figure 4 and in the text is that approx. half of the dogs from the SRB group moved to the Clear group in October.	1	2	ani12040482_perova	1
+Leaving time has been changed throughout to ‘time left alone’ L33 10th instead of “10th” Thank you, we’ve corrected the typo.	2	1	L22 and elsewhere: dogs’ “leaving time” or “leaving hours” I think this expression is misleading because it is not the dogs that have left home, but the owners. But this is the opinion of a person using English as a second language.	1	2	ani12040482_perova	1
+Thank you for pointing this out. We define SRB in the opening lines of the Introduction (45-50), however you are right that in places the terms can get confusing. Section 3.7.3. details behaviours owners reported as associated with separation, that are not what class as SRBs. Those were names ‘other’ separation behaviour but this has now been clarified, hopefully, by remaining the section ‘Other’ separation-linked behaviour, and then in the text it is now stated they are not SRBs. The term ‘separation anxiety’ was used in the introduction where it referenced a study that used that specific term. This has now been put into quote marks to be clear it is their term, not ours. The one mention of separation-distress has also now been changed.	2	1	L45 Separation behaviour, separation-related behaviour, separation distress, separation anxiety terms are used interchangeably throughout the text, although these terms do not necessarily describe the same behaviours. Please provide a clear definition of the behaviour aimed to study and stick to one term.	1	2	ani12040482_perova	1
+Thank you, we’ve corrected the typo.	2	1	L92 delete the comma after Bowen et al.’s.	1	2	ani12040482_perova	1
+This has now been discussed further in lines 98-100.	2	1	L92-92 28.5% of dogs reported to have SRPs – this is a much higher number than mentioned in previous studies and needs to be discussed.	1	2	ani12040482_perova	1
+This has now been moved to the beginning of the methods section.	2	1	L118 Materials and Methods This section (after the Ethics) should begin with the Subjects section. It is confusing to read about the surveys without knowing the sample sizes per survey and the demographics.	1	2	ani12040482_perova	1
+Agreed, this has been amended and condensed, thank you.	2	1	L127-137 The dates of data collections have been already mentioned in L121-126, so there is no need for repeating them. The whole section is not relevant to the main question.	1	2	ani12040482_perova	1
+Thank you, this seemed ok until the manuscript was uploaded but we’ve corrected it now.	2	1	L131-132 and elsewhere: (Error! Reference source not found.) This error occurs 7 times in the text, please correct.	1	2	ani12040482_perova	1
+We asked about the dogs’ management and behaviour over the previous 7 days so as to limit recall bias and anchor the data to a specific snapshot of time, rather than being more general. This was first asked for the first survey which was specifically looking at changes in management and behaviour during the strict lockdown period, so needed to be time specific. Questions were then repeated in the follow-up survey. Wording has been added to lines 165-169 now to clarify this.	2	1	L148 Please explain why a period of 7 days were asked to be reported	1	2	ani12040482_perova	1
+this has been done now.	2	1	L165-167 Figure 1. This Figure should be moved to the Supplement material as it tells nothing about the dogs’ behaviour.	1	2	ani12040482_perova	1
+Thank you for spotting this, we have updated both the methods (L243-247) and results section (L370-374) with further detail of inter-rater reliability checks.	2	1	L216-218 “The text was then coded by two researchers” I could not find agreement data reported later.	1	2	ani12040482_perova	1
+At the end of survey 1 there was the option for people to provide their demographic details, which not everyone did. This was done to conform to GDPR by asking only the most necessary questions. Given that the research question here is related to the dogs’ behaviour change over time versus leaving hours, it wasn’t deemed relevant to include analysis of the owner demographics.	2	1	L238 Do the authors have information about the Owner demographics? What was the proportion of females? Was there a relationship between age and leaving hours? Or explain why this information has not been used.	1	2	ani12040482_perova	1
+This has now been done.	2	1	L232-242 These sections should be moved to the Materials and Methods as a “Subjects” section	1	2	ani12040482_perova	1
+For the purpose of this study it was required that we had baseline data on the dogs’ leaving behaviour so that we could classify them as SRB+ or – before the pandemic began. Owners of dogs who were not left alone in a typical week in February, were not asked about their dogs’ separation-related behaviour, so we did not have these data for them which is why they were excluded. New text has been added to the Subjects section of the methodology to explain how many owners answered the follow up questionnaire, the response rate and what percentage and number of dogs were not left home alone before the pandemic. The new section beginning at line 146 now reads: “A total of 2,425 people completed the follow-up survey for the same dog they scored in the original survey (a 51.9% response rate), of which 2,285 had previously provided data to indicate whether their dog was left alone or not in February 2020. Of these, 15.3% dogs (n=305) were not left alone at all in an average week in February, and as such, these dogs did not have baseline data for their behaviour when left so were excluded from further analysis.” Additionally, the number of people we emailed the invitation to (n=4,670) has been added to line 139.	2	1	L232 How many owners have filled in the questionnaire altogether? What was the proportion of owners who did not leave their dog at home before the pandemic? Why were they excluded? It is perfectly feasible that some owners did not leave their homes because they were for example, ill, but they did so during the pandemic. What was the proportion of owners who indicated that they would like to participate in the follow-up study? Among them, what was the proportion of owners who has not responded to the request afterward?	1	2	ani12040482_perova	1
+For the 1407 dogs who were clear of SRB in February, they changed from being 100% clear to being 90.1% clear, with 1 in 10 of these dogs developing SRB they didn’t have previously. However, because of the dramatic improvement seen in the dogs that had SRB before the pandemic, the overall prevalence of SRB in the population decreased from 22% in February, to 17.2% in October (reported in the paragraph above). The numbers have been clarified in this section to help future readers, and additional discussion has been added around the overall decrease in SRB prevalence in both the abstract and in the discussion (lines 524-544).	2	1	L278-280 “There were 1,407 dogs who were clear of SRB at baseline in February 2020. When looking at SRB status in October, of 1,187 dogs who were left alone in October, 117 (9.9%) were reported to have shown at least one SRB”. In other words, 1407/1807 (78%) of dogs were clear of SRB in February and 90.1% in October. This is good news, and it is unclear for me why is it reported as bad news. The next section (l281-283) emphasizes that 55.7% of the dogs who were reported to show SRB in February were clear in October. This is a big improvement, I think.	1	2	ani12040482_perova	1
+Thank you, we’re glad you liked it! Further explanation to help interpret the figure has now been added.	2	1	L285 Figure 4. This is a great figure, thank you for including it. However, an explanation would be useful for those readers who are not familiar with this type of depicting.	1	2	ani12040482_perova	1
+Thank you for spotting that!	2	1	L291 The odds ratio (OR) is 5.38 and not 4.38 in Table 1.	1	2	ani12040482_perova	1
+This was explained further down in the section on the multi-variable model, but I have now moved the explanation to here, where it’s first mentioned and added some additional text also.	2	1	L293-295 “The final variable associated with October SRB status was the difference in the number of days dogs were left alone for between the February baseline and lockdown”.	1	2	ani12040482_perova	1
+Thank you, this has been added now.	2	1	OR was 0.81 here. Please explain the relationship between the variables.	1	2	ani12040482_perova	1
+Thank you! Looks like I read the z-value when I was typing. I promise this has been proof read, but it’s so easy to miss things, thank you so much!	2	1	L298 Table 1 legend: Add what bold indicates L307 The odds ratio (OR) is 4.97 in Table 1 and not	1	2	ani12040482_perova	1
+Since there were no non-significant variables it wasn’t necessary to distinguish the results.	2	1	L307 The odds ratio (OR) is 4.97 in Table 1 and not 3.60 Thank you!	1	2	ani12040482_perova	1
+This has been corrected, thank you.	2	1	L314 Table 2: bold is not used here although it was used in Table 1	1	2	ani12040482_perova	1
+Thank you for this, we were familiar with the paper but had missed that link. New text has been added here now to discuss this.	2	1	L333 table 3: same: bold is not used here	1	2	ani12040482_perova	1
+Agreed, this has been explicitly stated here now.	2	1	L382 Please refer to Flannigan and Dodman (2001) here (owner’s work schedule affects separation problems). Flannigan, G., & Dodman, N. H. (2001). Risk factors and behaviors associated with separation anxiety in dogs. Journal of the American Veterinary Medical Association, 219(4), 460-466.	1	2	ani12040482_perova	1
+Thank you so much for your time and these comments, they have really helped to strengthen the paper and we really appreciate it.	2	1	L395-397 “This value is likely to be an underestimate” The underestimation has already occurred during the baseline.	1	2	ani12040482_perova	1
+In order to address your feedback fully, we felt it necessary to split Figure 7. The data contained in the original Figure 7 is now split between Figure 7 (deals with CD14 and CD68 only) and Figure 8 (deals with CD11b and CD36).	2	1	Figure 7b is not very credible, when U937 become macrogphages adhere to the plate and assume the morphology of a macrophage. the figure is not clear and not well visible.	1	2	antiox11040683_makarova	1
+Fig 7b. We have removed the Giemsa images and replaced with phase contrast images to better show cell morphology under our light microscope. We also include representative flow cytometric overlayed histograms, which display CD14 and CD68 expression changes between our Vector control and WT / CXXS BCAT1 cells. Included on these histograms is the mean fold MFI change (ratio).	2	1	The authors should also show the flow cytometers panels related to cd68 and cd14.	1	2	antiox11040683_makarova	1
+Fig S4B. This is fully compensated.	2	1	Small comment, the supplementary Fig. S4B would need some compensation work....this would be better to correct before final approval for publication ( up to editor).	1	2	antiox11040683_makarova	1
+Fig 4d. This is going in the correct direction.	2	1	Also the gating strategy on Fig. 4d ( is going in right direction), what I am missing here is the exclusion of dead cells?).	1	2	antiox11040683_makarova	1
+Thank you for taking time to review are work and for giving us the opportunity to comment on the use of U937 cells as a limitation for the work presented. We have included a section dedicated to addressing this in the discussion section, along with supporting references.	2	1	The data presented by the authors are robust and interesting but are based on a single AML cell line, which is not a bonafide AML cell line (derived from the pleural effusion of a 37-year-old, White, male patient with histiocytic lymphoma, according to ATCC).	1	2	antiox11040683_makarova	1
+"We agree that transient analysis is important in designing a robot joint. However, the transient analysis requires relevant parameters of the robot arm be determined besides the parameters of the joint. The following sentence has been added at the end of Conclusions: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors"""	2	1	Transient analysis followed by reliability experiments/analysis/modeling of the proposed structure would be interesting and critical in aspect of joint application contrains. In my opoinion is should be added or explained in context of scheduled follow-up activities.	1	2	app12052642_perova	1
+The mentioned reference is focused on an analytical method for modeling large deflections in compliant mechanisms, while this work presents a method of enhancing the tensile strength of lamina emergent joints by using double-layer materials. As compared to Ref. [19] which focuses on concepts and demonstrations, this work mainly focuses on analytical modeling of one design concept and provides finite element and experimental validations. The following sentences have been added	2	1	The article being evaluated is an extension of the above cited reference, and the authors would be well advised as to summarize this article, in part, and to delineate what has changed as of 2019, i.e. put this in a paragraph and to explain the subsequent evolution since 2019.	1	2	app12052642_perova	1
+"As to the second comment, Ref. [34] has been new added in the reference list and the following sentence has been added at the end of the second paragraph of Introduction: ""Ref. [34] made a robot using double-layer LEMs, which is driven by magnetic force. This paper further discussed how to adjust the motion by changing different parameters of the mechanism"". The following sentence has been added at the end of Conclusion: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors."""	2	1	Secondly what is missing in the article is in terms of application a clear delineation as to what benefits robotics accrue by the developments summarized in this article . What new capacities and abilities so created by this evolutionary jump Aside from that the article is solidly researched and is useful as a source of technology development	1	2	app12052642_perova	1
+We have rewritten the conclusions and abstract, and added additional keywords.	2	1	Keywords are not sufficient after the abstract.	1	2	app12052642_perova	1
+"at the beginning of the last paragraph of Introduction: ""In this work, based on the design concept of improving the tensile stiffness of LET joints in Ref. [19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint. A theoretical model is derived for the DL-LET joint and verified by FEA and experimental results."" The following sentences have been added in the first paragraph of Introduction to mention the base paper and the related work (more references have been added): ""Finding suitable compliant joints [7] can be a key step in designing LEMs. Jacobsen et al. [8] first proposed the design concept of utilizing torsional deflection to obtain out-of-plane motions from planar structures, leading to a new group of compliant joints for LEMs called Lamina Emergent Torsional (LET) joints""."	2	1	Work relevant to the research topic is not incorporated, i.e., the work done by other researchers / authors in this field are not incorporated.	1	2	app12052642_perova	1
+The references have been reformatted according to standard journal format.	2	1	References are not written in the standard journal format.	1	2	app12052642_perova	1
+We have added more relevant references in this version.	2	1	Work relevant to the research topic is not incorporated, i.e., the work done by other researchers / authors in this field are not incorporated.	1	2	app12052642_perova	1
+The contribution of this work with respect to the relevant work is further described in Introduction (as marked in the manuscript).	2	1	Please add more details regarding paper’s novelty, it is not very clear what are the novelties of this paper.	1	2	app12052642_perova	1
+The theoretical part has been extended in the modeling section (page 3).	2	1	Please add more details of how the theory from the first sections is applied in the results section and better detail the theoretical part, in this version the theoretical part is almost inexistent.	1	2	app12052642_perova	1
+The modeling section (page 3) has been extended to make it clear how the parameters were obtained.	2	1	Please detail how the parameters were obtained.	1	2	app12052642_perova	1
+The recommended and other relevant references have been added.	2	1	The state of the art it is very poor regarding representative papers, maybe the author could add the following publications: o Hybrid Data-Driven Fuzzy Active Disturbance Rejection Control for Tower Crane Systems, European Journal of Control, vol. 58, pp. 373-387-11, 2021. o Event-Triggered Adaptive Fuzzy Control for Stochastic Nonlinear Systems with Unmeasured States and Unknown Backlash-Like Hysteresis, IEEE Transactions on Fuzzy Systems, doi 10.1109/TFUZZ.2020.2973950, pp. 1–19, 2020.	1	2	app12052642_perova	1
+Subsection 3.2 has been rewritten to add more details regarding the obtained results.	2	1	Please add more details regarding the obtained results.	1	2	app12052642_perova	1
+The following sentence has been added at the end of first paragraph of Conclusions to clarify disadvantage of DL-LET: “The DL-LET joint is vulnerable to delamination of the two layers.”	2	1	Add the both the advantages and the disadvantages of the proposed method. In the proposed manuscript only the advantages are presented.	1	2	app12052642_perova	1
+Thank you for your explanation.	3	2	"We agree that transient analysis is important in designing a robot joint. However, the transient analysis requires relevant parameters of the robot arm be determined besides the parameters of the joint. The following sentence has been added at the end of Conclusions: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors"""	2	1	app12052642_perova	1
+The authors answered to all my concern in the second version of the paper. In conclusion the paper deserves to be acceped as contribution in Applied Science journal.	3	2	The following sentence has been added at the end of first paragraph of Conclusions to clarify disadvantage of DL-LET: “The DL-LET joint is vulnerable to delamination of the two layers.”	2	1	app12052642_perova	1
+We have added all the new references suggested by Referee 2 on page 3 of the new manuscript. These includethe ones mentioned in our reply to Referee 1’s point 4 above, and all also the references: Corney98,Mekhov09,leroux10,Mekhov12,Niedenzu13,Lee14.	2	1	Other examples of many-atom-cavity continuous observation papers: W. Niedenzu, S. Schütz, H. Habibian, G. Morigi, and H. Ritsch, Phys. Rev.  88, 033830 (2013). I. B. Mekhov and H. Ritsch, Phys. Rev. Lett. 102, 020403 (2009). J. F. Corney and G. J. Milburn, Phys. Rev. A 58, 2399 (1998). I. D. Leroux, M. H. Schleier-Smith, and V. Vuletić, Phys. Rev. Lett. 104, 073602 (2010).	1	2	atoms4010002_makarova	1
+Concerning the referee’s other point, about the suitability of this system to measure weak forces, we first note that the use of Bloch oscillations without a cavity is proving to be an excellent way to measure gravity (e.g. the experiments by G.M. Tino’s group).	2	1	The role of sensing may in the end be less important and could be downplayed a bit in the manuscript.	1	2	atoms4010002_makarova	1
+Concerning the question of whether we have taken the effects of higher Bloch bands into effect, our numerical calculations do take this into account. The question of higher Bloch bands is a little subtle when it comes to our analytical calculation done in the Wannier-Stark picture. In the presence of the force, the true eigenstates of the system are no longer the Bloch states but the localized Wannier-Stark states. In the analytical calculation we restrict ourselves to just the ground band Wannier-Stark states. However, even with this approximation we find very good agreement between the numerical and analytical calculations.	2	1	What is the role of excitations to higher Bloch bands?	1	2	atoms4010002_makarova	1
+Done. We have re-written the first part of the abstract as: “In this paper we give a new description, in terms of optomechanics, of previous work on the problem of an atomic Bose-Einstein condensate interacting with the optical lattice inside a laser-pumped optical cavity and subject to a bias force such as gravity \cite{Ven09,Ped09,Gol14}. An atomic wave packet in a tilted lattice undergoes Bloch oscillations; in a highfinesse optical cavity the backaction of the atoms on the light leads to a time-dependent modulation of the intracavity lattice depth at the Bloch frequency which can in turn transport the atoms up or down the lattice.”	2	1	(1) It should be somehow mentioned more clearly (maybe in the abstract) that the idea is not completely new but work presents a new and more intutive picture of the system dynamics.	1	2	atoms4010002_makarova	1
+We have added the following sentences on p3 (and four new references, including the one to Gangl’s work, in the bibliography): “A number of experiments have already demonstrated how the light transmitted by a cavity can be used to track the motion of atoms trapped inside \cite{Hood98,Hood00,Pinkse00}, and in particular, a theoretical analysis of the information stored in the frequency spectrum has been given in \cite{Gangl00}, showing that atomic motion introduces sidebands either side of the pump frequency. In our case, Bloch oscillations at angular frequency $\omega_{B}$ generates sidebands separated from the pump frequency by $\pm \omega_{B}$ (and harmonics thereof in the strong coupling regime).Because the Bloch frequency is proportional to the applied force $\omega_{B}= F d/\hbar$, where $d=\lambda/2$ is thelattice period, a detection of the spectrum of the transmitted light gives $F$ directly.”	2	1	(2) It is very helpful to connect cavity output spectra and forces. To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40. This should be mentioned.	1	2	atoms4010002_makarova	1
+Actually, we do not claim that oscillation frequencies in general will be unaffected by the backaction. Rather, we claim that one particular frequency, the Bloch frequency, will be unaffected.	2	1	(3) The authors claim there should be no backaction of the field on the oscillation frequencies. This is somehow in contradiction with the eigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.	1	2	atoms4010002_makarova	1
+Concerning the change in wavelength (and hence change in lattice constant) due to the refractive index of the gas, this effect is tiny. We consider it in section 6, on p17 and show that the correction to the refractive index is of order 10^-9 for our parameters. This is to be distinguished from the effective change in the cavity length which, due to the resonance effect, is substantial and is precisely what lies behind the amplitude modulation of the lattice.	2	1	Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.	1	2	atoms4010002_makarova	1
+Yes we agree that there will be extra noise due to the open nature of the cavity, and we already refer to this on p3 of the Introduction: “The disadvantage of working in a cavity is that quantum measurement backaction, in the form of random fluctuations in the cavity field due to photons spontaneously leaking out of the cavity, heats up the cold atoms and limits the coherence time of the measurement \cite{Ven13}.” where the cited paper is one of ours [Prasanna Venkatesh, B.; O’Dell, D. H. J.; Bloch oscillations of cold atoms in a cavity: Effects of quantum noise. Phys. Rev. A 2013, 88, 013848] where we directlycalculated the heating effect for Bloch oscillating atoms in a cavity within the Bogoliubov approximation.	2	1	(a) any measurement of the output field will generate backaction onto the condensate (Mekhov, J. Phys. B: Atomic, Molecular and Optical Physics, 45(10), 102001). This will generate extra noise, but might also help as it reduces atom number uncertainty in the cavity.	1	2	atoms4010002_makarova	1
+The coherence time for the many-particle Bloch oscillating state is in fact very hard to calculate (mainly because even the coherent part is time-dependent rather than stationary). It can, however, be estimated and so we have added in the following sentences on p3:“\textcolor{red}{The coherence time is particularly hard to calculate in the Bloch oscillating case \cite{Ven13} due to the time dependence introduced by the Bloch oscillations, especially in the presence of many particles, but it can be roughly estimated to be $\tau=\tau_{\mathrm{sp}}/(1+C)$ \cite{Ven09} at cavity resonance, where$\tau_{\mathrm{sp}}^{-1}=2 \gamma \vert \alpha \vert^2 \Omega_{0}^2/\Delta_{a}^2$ is the spontaneous emission rate at an antinode. The factors $\vert \alpha \vert^2$ and$\Delta_{a}$ are the mean number of cavity photons and the detuning of the laser from atomic resonance, respectively, and will be properly defined in the next section. The numerical value of $\tau$ for the parameters considered in this paper will be given in Section \ref{sec:metrology}. Of course, Bose-Einstein condensates can be continuously measured and used for sensingwithout a cavity, e.g.\ \cite{Ruostekoski98,Dalvit02,Saba05,Lee12,Java13}, but the cavity case is particularly interesting because it allows for a strong atom-light interaction even in the quantum regime.} “In addition, in Section 6 (paragraph 4, Pg 18) we have provided further details required to estimate the coherence time for the parameters used in the paper. We find thatcoherence times of up to a couple of seconds (many thousands of Bloch oscillation periods) are possible	2	1	(b) There will be heating limiting the available measurement time.	1	2	atoms4010002_makarova	1
+We have added all the new references suggested by Referee 2 on page 3 of the new manuscript. These includethe ones mentioned in our reply to Referee 1’s point 4 above, and all also the references: Corney98,Mekhov09,leroux10,Mekhov12,Niedenzu13,Lee14.	2	1	There are other papers that link BECs, sensing and continuous quantum measurement process (in the absence of a cavity) M. Saba, T. A. Pasquini, C. Sanner, Y. Shin, W. Ketterle, and D. E. Pritchard, Science 307, 1945 (2005).	1	2	atoms4010002_perova	1
+Concerning the referee’s other point, about the suitability of this system to measure weak forces, we first note that the use of Bloch oscillations without a cavity is proving to be an excellent way to measure gravity (e.g. the experiments by G.M. Tino’s group). The sensitivity of their measurements, after many years of development, are only one order of magnitude above our estimated sensitivity, and their measurements take one hour whereas ours should take one second which means it might have applications in rather different situations (e.g. local gravity mapping while flying). Our work is very much still in progress and this paper will not, we hope, be our final word on this subject. Indeed, we point out a possible way forward in the last section that builds on the insight gained from the optomechanics analogy. It seems likely that the homodyne phase measurement regime, where backaction can be reduced, will likely be more favourable when it comes to absolute sensitivity than photon counting type measurement we focus on at the moment. For these reasons we would like to keep the measurement aspects of the paper intact: we would like to advertise them and, perhaps, be a little provocative.	2	1	The link to the sensing of weak forces (in this case gravity) is interesting, but I am not convinced whether this is a practical approach. The estimated accuracy is $10^6$ that does not sound great.	1	2	atoms4010002_perova	1
+Concerning the question of whether we have taken the effects of higher Bloch bands into effect, our numerical calculations do take this into account. The question of higher Bloch bands is a little subtle when it comes to our analytical calculation done in the Wannier-Stark picture. In the presence of the force, the true eigenstates of the system are no longer the Bloch states but the localized Wannier-Stark states. In the analytical calculation we restrict ourselves to just the ground band Wannier-Stark states. However, even with this approximation we find very good agreement between the numerical and analytical calculations.	2	1	What is the role of excitations to higher Bloch bands?	1	2	atoms4010002_perova	1
+Concerning the question of radiationpressure forces, we take them into account on the atoms but not on the mirrors.	2	1	Have the effects of radiation pressure forces, etc., taken into account?	1	2	atoms4010002_perova	1
+Done. We have re-written the first part of the abstract as: “In this paper we give a new description, in terms of optomechanics, of previous work on the problem of an atomic Bose-Einstein condensate interacting with the optical lattice inside a laser-pumped optical cavity and subject to a bias force such as gravity \cite{Ven09,Ped09,Gol14}. An atomic wave packet in a tilted lattice undergoes Bloch oscillations; in a highfinesse optical cavity the backaction of the atoms on the light leads to a time-dependent modulation of the intracavity lattice depth at the Bloch frequency which can in turn transport the atoms up or down the lattice.”	2	1	(1) It should be somehow mentioned more clearly (maybe in the abstract) that the idea is not completely new but work presents a new and more intutive picture of the system dynamics.	1	2	atoms4010002_perova	1
+We have added the following sentences on p3 (and four new references, including the one to Gangl’s work, in the bibliography): “A number of experiments have already demonstrated how the light transmitted by a cavity can be used to track the motion of atoms trapped inside \cite{Hood98,Hood00,Pinkse00}, and in particular, a theoretical analysis of the information stored in the frequency spectrum has been given in \cite{Gangl00}, showing that atomic motion introduces sidebands either side of the pump frequency. In our case, Bloch oscillations at angular frequency $\omega_{B}$ generates sidebands separated from the pump frequency by $\pm \omega_{B}$ (and harmonics thereof in the strong coupling regime).Because the Bloch frequency is proportional to the applied force $\omega_{B}= F d/\hbar$, where $d=\lambda/2$ is thelattice period, a detection of the spectrum of the transmitted light gives $F$ directly.”	2	1	(2) It is very helpful to connect cavity output spectra and forces. To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40. This should be mentioned.	1	2	atoms4010002_perova	1
+Actually, we do not claim that oscillation frequencies in general will be unaffected by the backaction. Rather, we claim that one particular frequency, the Bloch frequency, will be unaffected. The Horak paper shows that Bloch energies (i.e. eigenmodes for a particle in a periodic potential…not to be confused with the Bloch oscillation frequency) will be changed by the backaction. This is because the presence of atoms in the cavity shifts the cavity resonance and hence the depth of the intracavity lattice: the Bloch energies are dependent on the potential depth. This change in potential depth due to backaction is fully included in our treatment. However, a change in the depth of a periodic potential does not change the Bloch frequency because this depends on the lattice’s spatial period, not the depth. Because this key aspect of our treatment can clearly lead to confusion we have modified the following sentence in the conclusions on p19, and added the reference to Horak’s paper: “Chief among these is that the backaction does not alter the frequency of the Bloch oscillations. By contrast, in the harmonic oscillator case there is the so-called optical spring effect which gives a dependence of oscillator frequency on field amplitude and detuning. To be clear, other motional frequencies are altered: because the intracavity lattice depth is modulated by the backaction this will affect certain types of atomic motion, for example the oscillation frequency of an atom about the bottom of one of the potential minima \cite{Horak00}. Nevertheless, the Bloch oscillation frequency is robust against this depth modulation because it only depends on the lattice period, not its depth.” Concerning the change in wavelength (and hence change in lattice constant) due to the refractive index of the gas, this effect is tiny. We consider it in section 6, on p17 and show that the correction to the refractive index is of order 10^-9 for our parameters. This is to be distinguished from the effective change in the cavity length which, due to the resonance effect, is substantial and is precisely what lies behind the amplitude modulation of the lattice.	2	1	(3) The authors claim there should be no backaction of the field on the oscillation frequencies. Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil. This is somehow in contradiction with the eigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.	1	2	atoms4010002_perova	1
+Yes we agree that there will be extra noise due to the open nature of the cavity, and we already refer to this on p3 of the Introduction: “The disadvantage of working in a cavity is that quantum measurement backaction, in the form of random fluctuations in the cavity field due to photons spontaneously leaking out of the cavity, heats up the cold atoms and limits the coherence time of the measurement \cite{Ven13}.” where the cited paper is one of ours [Prasanna Venkatesh, B.; O’Dell, D. H. J.; Bloch oscillations of cold atoms in a cavity: Effects of quantum noise. Phys. Rev. A 2013, 88, 013848] where we directlycalculated the heating effect for Bloch oscillating atoms in a cavity within the Bogoliubov approximation. Following the referee’s suggestion we have added the reference to Mekhov’s paper.The coherence time for the many-particle Bloch oscillating state is in fact very hard to calculate (mainly because even the coherent part is time-dependent rather than stationary). It can, however, be estimated and so we have added in the following sentences on p3:“\textcolor{red}{The coherence time is particularly hard to calculate in the Bloch oscillating case \cite{Ven13} due to the time dependence introduced by the Bloch oscillations, especially in the presence of many particles, but it can be roughly estimated to be $\tau=\tau_{\mathrm{sp}}/(1+C)$ \cite{Ven09} at cavity resonance, where$\tau_{\mathrm{sp}}^{-1}=2 \gamma \vert \alpha \vert^2 \Omega_{0}^2/\Delta_{a}^2$ is the spontaneous emission rate at an antinode. The factors $\vert \alpha \vert^2$ and$\Delta_{a}$ are the mean number of cavity photons and the detuning of the laser from atomic resonance, respectively, and will be properly defined in the next section. The numerical value of $\tau$ for the parameters considered in this paper will be given in Section \ref{sec:metrology}. Of course, Bose-Einstein condensates can be continuously measured and used for sensingwithout a cavity, e.g.\ \cite{Ruostekoski98,Dalvit02,Saba05,Lee12,Java13}, but the cavity case is particularly interesting because it allows for a strong atom-light interaction even in the quantum regime.} “In addition, in Section 6 (paragraph 4, Pg 18) we have provided further details required to estimate the coherence time for the parameters used in the paper. We find thatcoherence times of up to a couple of seconds (many thousands of Bloch oscillation periods) are possible	2	1	(b) There will be heating limiting the available measurement time. as the time scale can be faster than om_recoil, it could limit the measurement time to a fairly short period. Some rough estimate on the time scales should be added. In particular for the blue detuned case not only the average momentum but also the momentum spread will grow.	1	2	atoms4010002_perova	1
+The review article by W Nakel and C. T. Whelan you refer to do indeed tackle the relativistic regime but at the expense of a huge amount of numerical work.	2	1	The authors did not mention this method for discussions in the manuscript. In earlier publications [Physics Reports 315 (1999) 409 and related references], a fully relativistic distorted-wave Born approximation (rDWBA) has been developed to describe the relativistic ionization processes.	1	2	atoms4010010_makarova	1
+The work of W Nakel and C. T. Whelan is devoted to study heavy atoms but Our work is aimed to study analytically the hydrogen atom, we believe that such a comparison is impossible.	2	1	The RPWBA method in the manuscript should be compared with the previous rDWBA theory.	1	2	atoms4010010_makarova	1
+We are not in a position to compare with these works since they are devoted to heavy atoms.	2	1	Again in earlier publications [Physics Reports 315 (1999) 409 and related references], there are existing experiments of triple differential cross sections for relativistic K-shell ionization of Ag, Au, and so on. The authors did not mention these experimental works in the manuscript.	1	2	atoms4010010_makarova	1
+Points (i) and (ii) have been corrected and changed following to the referee’s amendments.	2	1	(ii) There are repeated Bibliography like ref. 11 and ref. 31.	1	2	atoms4010010_makarova	1
+A major revision of the paper has been carried out to take all the comments, suggestions and english corrections into account.	2	1	In addition, I do believe that several figures could be combined in order to minimize journal space.	1	2	atoms4010010_makarova	1
+The changes is made in the text.	2	1	"Page 9:* ""experimental one"" -> ""experimental ones"""	1	2	atoms4010010_makarova	1
+The review article by W Nakel and C. T. Whelan you refer to do indeed tackle the relativistic regime but at the expense of a huge amount of numerical work. So, we think that presenting for the first time a work that incorporates fully analytical results and QED formalism is interesting to the scientific community. The work of W Nakel and C. T. Whelan is devoted to study heavy atoms but Our work is aimed to study analytically the hydrogen atom, we believe that such a comparison is impossible. Another difference that makes comparison difficult is that the work of W Nakel and C. T. Whelan uses coplanar asymmetric geometry while in our work we use coplanar symmetric geometry.	2	1	The authors did not mention this method for discussions in the manuscript. The RPWBA method in the manuscript should be compared with the previous rDWBA theory. At least the authors should mention the differences of the two methods and how far the RPWBA can be compared to rDWBA? In earlier publications [Physics Reports 315 (1999) 409 and related references], a fully relativistic distorted-wave Born approximation (rDWBA) has been developed to describe the relativistic ionization processes.	1	2	atoms4010010_perova	1
+We are fully aware that the mainstream trend is to study heavy atoms. But some fundamental processes have not until now been investigated fully and analytically and it is our very humble belief that such a fundamental work must be presented to the international scientific community. We are not in a position to compare with these works since they are devoted to heavy atoms.	2	1	Again in earlier publications [Physics Reports 315 (1999) 409 and related references], there are existing experiments of triple differential cross sections for relativistic K-shell ionization of Ag, Au, and so on. The authors did not mention these experimental works in the manuscript. It would be nice if the RPWBA method in the manuscript can be compared with the experiments.	1	2	atoms4010010_perova	1
+Points (i) and (ii) have been corrected and changed following to the referee’s amendments.	2	1	The authors have not carefully proofread this manuscript. Evidences of this includes (i) some errors in English typing, e.g. line 27 and line 119: it should be ‘plane wave …’ instead of ‘plan wave …’. Line between 33 and 34: it should be ‘The scattered positron and ejected electron …’ instead of ‘The scattered electron and ejected positron …’. (ii) There are repeated Bibliography like ref. 11 and ref. 31.	1	2	atoms4010010_perova	1
+As you requested, we have made all necessary changes in our manuscript. A major revision of the paper has been carried out to take all the comments, suggestions and english corrections into account. We believe that the paper has been significantly improved.	2	1	In addition, I do believe that several figures could be combined in order to minimize journal space.	1	2	atoms4010010_perova	1
+The changes is made in the text.	2	1	"Page 9:* ""experimental one"" -> ""experimental ones"""	1	2	atoms4010010_perova	1
+We added references to the Wolf and Tourrencin and A. Peters, K.Y. Chung and S. Chu articles. The latter of these articles does not address the quantum corrections 2.	2	1	The first example is the result announced in Equation (4), concerning the phase shift due to the gravity gradient gamma. This effect was first calculated by Wolf and Tourrencin Physics Letters A 251 (1999) pp 241–246 and discussed with more details in A. Peters, K.Y. Chung and S. Chu, Metrologia 2001, 38, 25–61.	1	2	atoms4020014_perova	1
+The referee implies that the leading term is not a quantum correction owing to the cancellation of a numerator proportional to hbar and a denominator in the spatial phase factor that is also proportional to hbar. This is, is some respect, a question of semantics, and has a long history in discussions of quantum effects in atom interferometry. Basically this term is a Doppler shift, which is clearly of classical origin, with no need to impose arguments related to cancellations of hbars in numerators and denominators. In the approach we follow using the Wigner representation, there are no numerators and denominators; the leading term has no hbar.	2	1	Moreover, the authors call this effect a quantum correction probably because it is proportional to hbar but it would be more clear to explain that, the leading term of the gravitation phase being independent of hbar because of a cancellation (hbar appears in the numerator and denominator of the quantity giving the phase), the term linear in gamma is also proportional to the recoil velocity which itself is proportional to hbar.	1	2	atoms4020014_perova	1
+The density matrix in the Wigner representation has been widely used in problems related to the recoil effect and laser cooling. For some mysterious reason it hasn’t used in much in problems involving atom interferometry. Of course there are hundreds of articles (maybe more) and books on the Wigner representation; however here we are concerned mainly with its application to calculations involving atom interferometry. The only examples we know of are the ones referred to in the paper.	2	1	A second example is the equation 9b for Wigner function: the associated quotation is ref. 17 by one of the authors and M.A. Kasevich in Phys Rev A (2006). A brief search on the web proves that a similar equation was used by Hongyi Fan Phys Rev A 656, 064102 (2002) who quotes two references dating from 1984 and a book published in 1994. I am not sufficiently aware of the use f Wigner function but I am sure that there are better and more useful references to quote in addition to ref 17!	1	2	atoms4020014_perova	1
+The article by Fam is devoted to the use of Wigner representation for the two-particle density matrix, which is outside the scope of our article. Moreover, in contrast to that article, we do not use an expansion of the equation for the Wigner distribution in powers of hbar. For both these reasons, we do not include a reference on this article 5.	2	1	A second example is the equation 9b for Wigner function: the associated quotation is ref. 17 by one of the authors and M.A. Kasevich in Phys Rev A (2006). A brief search on the web proves that a similar equation was used by Hongyi Fan Phys Rev A 656, 064102 (2002) who quotes two references dating from 1984 and a book published in 1994. I am not sufficiently aware of the use f Wigner function but I am sure that there are better and more useful references to quote in addition to ref 17!	1	2	atoms4020014_perova	1
+As far as we can tell, the phi_Q term is not related directly to the spreading of the wave-packet. It arises as a quantum correction to phase factors during periods of free evolution. At this point, we do not have any other physical interpretation. We do not know yet how this term would manifest itself in the other approaches (path-integrals, ABCD theorem and etc.). This remains an interesting topic for future work, but is outside the scope of our article.	2	1	In my opinion, phi_Q is the real quantum correction due to the spreading of the wavepacket during propagation. If I am correct, this should be explained.	1	2	atoms4020014_perova	1
+We added a paragraph re-emphasizing the choice of x_C, x_qC and x_QC just after Equation (99); these terms do not have to the same leading terms in T^2.	2	1	The meaning of the three x_C, x_qC and x_QC (Equation (99)) is far from obvious but it is very surprising that the leading term in T^2 is different?	1	2	atoms4020014_perova	1
+Several additional paragraphs have been added near the figures to help explain the plots appearing in these figures. For the stationary atom case, the limits on y_{m0} are determined from inequality (106) and it is indeed possible for y_{m0} to be less than y_{min}. The manner in which we arrived at the regions of validity of the approximations is now described in more detail.	2	1	The paper contains only one figure with 28 panels presenting the results of the calculations. I think that this figure is very difficult to read! I do not understand clearly why results are plotted with y < y_min if this not physical: please explain!	1	2	atoms4020014_perova	1
+In the article we obtained results that are specific for the parameters of the atom interferometer and test mass’ shape and trajectory. We think for the each case one should generate one figure 3 to answer all questions regarding the validity and role of different parts of the phase and approximate expressions for them. Instead of using a lot of dashed curves, we decided to use a panel of figures. Although each plot is difficult to read at “normal” magnification, the online reader can easily magnify any plot when accessing the article in PDF format. We have added a note to this effect.	2	1	The paper contains only one figure with 28 panels presenting the results of the calculations. I think that this figure is very difficult to read! I do not understand clearly why results are plotted with y < y_min if this not physical: please explain!	1	2	atoms4020014_perova	1
+We added a schematic figure of the geometry of our problem.	2	1	A figure explaining the geometry of the experiment would also help.	1	2	atoms4020014_perova	1
+In the paragraph after Equation (30) we added a list of requirements that will insure that the signal does not depend critically on the initial atomic state phase space distribution. These requirements minimize the corrections arising from diffraction to which the referee refers 11.	2	1	The process is calculated as if the gravitational forces can be neglected during the diffraction laser pulses. This is probably an excellent approximation but please comment it!	1	2	atoms4020014_perova	1
+Difficulties related to stray interferometers exist and are important, but are not addressed in our paper, as the referee points out. We are assuming that the effects can be minimized in a fountain geometry using a variety of experimental techniques. In any event, a discussion of these effects is beyond what we are able to discuss in this article.	2	1	The fountain is assumed to be exactly symmetric (see line after Equation (100)). It is well known that with symmetric fountains diffraction with retro-reflected laser beams produces stray interferometers... Minor Remarks	1	2	atoms4020014_perova	1
+The word “potential” has been changed to “acceleration” in the sentence after Equation (1).	2	1	I noted an error after Equation (1): ... the gravitational potential is constant... I assume that the authors meant acceleration g, not the potential.	1	2	atoms4020014_perova	1
+We know that this paper is “equation-rich”, but we have tried to provide some physical guidance for the reader.	2	1	The paper is quite difficult to read, because of very many bulky equations. I do not see how to reduce the number of equations but I think that some more comments would help the reader.	1	2	atoms4020014_perova	1
+A figure has been added to clarify the geometry of the problem. Moreover, additional discussion of the trajectories involved are now given in paragraphs that were added to help explain the plots given in Figures 2 and 3.	2	1	The relative location of the test mass to the cold atom cloud should be clearly stated during the calculation, as the gravitational force is a vector. If the test mass is not located in the direction of the evolution trajectory of the atom cloud, the applicability of some formulas (e.g. Equation (3)) need to be verified. It will be helpful to the readers if a figure is provided to show the locations.	1	2	atoms4020014_perova	1
+The wave function cannot be used when we know only some statistical properties of the initial phase space distribution.	2	1	The cloud is assumed to be characterized by a Wigner distribution function. What are the advantages to use Wigner distribution function instead of wave function?	1	2	atoms4020014_perova	1
+We thank the referee for pointing out this additional reference and have added it to the references.	2	1	In the Introduction, quite a lot of papers are cited concerning the applications of atom interferometer in varies fields. Just for the note of the authors, there are some new applications of atom interferometer, e.g., in the accurate measurement of the quadratic Zeeman coefficient [J. Phys. B: At. Mol. Opt. Phys., 47(1): 015001, 2014].	1	2	atoms4020014_perova	1
+As suggested by the reviewer, the non-pertinent references has been checked throughout the manuscript and unnecessary references are removed too.	2	1	The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.	1	2	biom12060834_makarova	1
+As suggested by the reviewer, now the title include the production optimization too and the new title is “Optimization and characterization of a novel exopolysaccharide from Bacillus haynesii CamB6 for food applications”.	2	1	Optimization of production constitutes an important part not mentioned in the title.	1	2	biom12060834_makarova	1
+As suggested by the reviewer, the acceptability for food application has now been mentioned with appropriate refences (Nicolaus et al., 2010; Kambourova et al., 2018; Gongi et al., 2022) in the revised version of the manuscript.	2	1	Acceptability for food applications was not discussed.	1	2	biom12060834_makarova	1
+According to reviewer´s suggestion, the language of the manuscript is thoroughly revised.	2	1	Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.	1	2	biom12060834_makarova	1
+As suggested by the reviewer, the consistent of the word had been checked throughout.	2	1	EPS vs. EPSs in plural form should be consistent throughout.	1	2	biom12060834_makarova	1
+Constant abbreviation had been checked throughout in the revised version of the manuscript.	2	1	Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.	1	2	biom12060834_makarova	1
+Accoding to the suggestion of the reviewer, Table 1 is revised and both coded – noncoded values are now included.	2	1	l. 160 There is no coded value in Table 1.	1	2	biom12060834_makarova	1
+Both the tables (table 1 and table 2) have been placed in the proper place where it expected in the revised manuscript.	2	1	l. 165 Table 2 where Table 1 expected	1	2	biom12060834_makarova	1
+The suggested correction has been done in the revised version of the manuscript	2	1	l. 169-170 thirty instead of thirteen ?	1	2	biom12060834_makarova	1
+The corrected has been made now.	2	1	l. 177 i instead of 0 in second term?	1	2	biom12060834_makarova	1
+The revision has been made as suggested.	2	1	l. 228 GPC defined on l. 231	1	2	biom12060834_makarova	1
+as suggested by the reviewer, PEG has been defined in the revised version of the manuscript	2	1	l. 230 PEG undefined	1	2	biom12060834_makarova	1
+The volume is now added in the revised version of the manuscript.	2	1	l. 247 Volume of 0.2 mM ethanolic DPPH solution?	1	2	biom12060834_makarova	1
+Suggestion has been included in the revised version of the manuscript throughout.	2	1	255, 286, and 441 et al.	1	2	biom12060834_makarova	1
+This to kindly mention to the reviewer, the map is the collection site of the EPS producing Bacillus haynesii CamB6. This is a first such report from this collection site, the map is necessary to keep in the manuscript.	2	1	Figure 1 What is the significance of the tick mark labels on the maps? Are these maps really necessary?	1	2	biom12060834_makarova	1
+Figure 2 has been modified as suggested by the reviewer.	2	1	Figure 2 No x-axis label Units not specified	1	2	biom12060834_makarova	1
+As suggested by the reviewer, Figure 3 is now modified with specified units of the axes and also in the figure legend.	2	1	Figure 3	1	2	biom12060834_makarova	1
+All the Suggestion has been included in the figure legend of revised version of the manuscript	2	1	417 and 432 30.0 instead of 3.0 ?	1	2	biom12060834_makarova	1
+To avoid the redundancy of the figures as suggested by the reviewer, figure 4 is removed from the revised version of the manuscript.	2	1	Units are not specified on the axes labels or in the legend. Figure 4 This figure is redundant as it gives the same information as Figure 3. Tick label values for glucose and yeast extract are different.	1	2	biom12060834_makarova	1
+As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript	2	1	433, 434, 454, 458, and 507 First section 3.6 (and its subsections) should be section 3.5.	1	2	biom12060834_makarova	1
+This has been checked now and revised.	2	1	l. 471, 600, 678, and 718 Reference format is different; corresponding references could be missing in the list.	1	2	biom12060834_makarova	1
+We performed 2D-NMR analysis , which allows us to determine exactly the α-Manp linkage, which is the major sugar component, and the β- structure of Galp and Glucp.	2	1	l. 480 Why was linkage analysis not performed?	1	2	biom12060834_makarova	1
+This is a heteropolysaccharide composed of 3 different sugars as determined for HPLC, which is highly common in this kind of polysaccharides isolated from bacterial strains (10.3390/foods11020156)	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	1
+There are several earlier report where different concentration of yeast extract is added to the culture media for optimized EPS production, and the resultant EPS also concisted of mannan.	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	1
+Several 2D-NMR spectra were performed to confirm 1H and 13C chemical shift of EPS structure, and their analysis was added to the manuscript.	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	1
+It was corrected in manuscript	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	1
+A new discussion of 1D and 2D NMR spectra was added and corrected.	2	1	l. 500 The chemical shift range is more than from 1 to 5. ll.	1	2	biom12060834_makarova	1
+According to the suggestion, section numbering has been checked thoroughly in the revised version of the manuscript.	2	1	l. 537 No section 3.6.2	1	2	biom12060834_makarova	1
+As suggested by the reviewer, the necessary revision in done in the manuscript.	2	1	l. 636 Glc and Gal	1	2	biom12060834_makarova	1
+As suggested by the reviewer, the information is corrected and revised now.	2	1	l. 683-684 Food-grade oils are not aliphatic and aromatic hydrocarbons.	1	2	biom12060834_makarova	1
+Suggestion has been included in the revised version of the manuscript	2	1	l. 725 possibility of future	1	2	biom12060834_makarova	1
+As suggested by the reviewer, minor errors throughout the manuscript has been revised now.	4	1	The English language was improved, but there are still errors.	3	2	biom12060834_makarova	1
+int. We agree with some of the comments that you made. However, we would like to remind you that the main purpose of this manuscript is not the complete elucidation of the EPS chemical structure but also the optimization of the EPS production, and characterization of the EPS functional properties. At the same time, we provided different chemical properties like: sugar identification and its molar ratio, molecular weight, functional group analysis and thermal stability of the EPS, and this journal has recently published other papers with similar or less chemical information on the EPS, for instance: Kuo, H.-C.; Liu, Y.-W.; Lum, C.-C.; Hsu, K.-D.; Lin, S.-P.; Hsieh, C.-W.; Lin, H.-W.; Lu, T.-Y. ; Cheng, K.- C. Ganoderma formosanum Exopolysaccharides Inhibit Tumor Growth via Immunomodulation. Int. J. Mol. Sci. 2021, 22, 11251. https://doi.org/10.3390/ijms222011251 Fetsiukh, A.; Conrad, J.; Bergquist, J.; Timmusk, S. Silica Particles Trigger the Exopolysaccharide Production of Harsh Environment Isolates of Growth-Promoting Rhizobacteria and Increase Their Ability to Enhance Wheat Biomass in Drought-Stressed Soils. Int. J. Mol. Sci. 2021, 22, 6201. https://doi.org/10.3390/ijms22126201.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+On the other side, we performed the 1D and 2D - NMR analysis as complementary information to the other techniques in the present work. Such analysis was performed to try to replace the traditional acid hydrolysis, methylation/acetylation and GC-MS used in glycoside analysis. On the opposite to NMR, we do not have access to this analysis, and we could not find it as a service.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+The HPLC and NMR results from the present work confirmed the heterogeneity of the EPS (3 different sugar moieties with different molar ratios), which agrees with the other reported mannan polysaccharides. The 1H and 13C NMR chemical shifts are comparable with literature reports of other similar EPS or oligosaccharides, where complex structures are identified by NMR too (Casillo et al., 2021; Chatterjee et al; 2018). Besides, our 2D-NMR analysis supported EPS heterogeneity, identifying different types of monomer linkages.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+Your mention of the non-uniform peak intensity confirming the three different sugar units found. This is because the signal intensity of NMR peaks is related to an -H abundance. Different monomer ratios are responsible for this variation and support our findings. Also, the 10 H1/H2 cross peaks observed in the COSY spectra supported our findings of EPS heterogeneity, given the following facts: 1) We found 3 different types of sugars by HPLC. 2) We identified by NMR at least 2 types of linkages α 1-2 Man-Man and alfa 1-4 Man-Man monosaccharides. 3) There are other 2 sugars for which the β- type of linkage was identified but not the exact sugar position. 4) Anomeric protons shifts from units at the reducing end of polysaccharide differs from the one in another position. In conclusion, there are different linkage possibilities (plus already identified) to different monosaccharides in different ring positions. This gives rise to a multiplicity of signals (not fully identified in this work), which could produce different H1/H2 correlations.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+About the COSY spectrum discussion, only a minor and partial discussion is presented that can be found below the Scheme in manuscript: “In the COSY spectrum, it was found its α-configuration by low-field H-1 signal at δ 5.34 and correlated with H-2 (δ 4.0), H-6 (δ 3.98 and 3.85), and H-5 (δ 4.12) (Speciale et al., 2022).”	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+Regarding the TOCSY and HMBC mentioned in the methods, they are now deleted from the manuscript, as suggested.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+Finally, we kindly request the reviewer to consider that this is the first approach to studying a Chilean hot spring EPS isolation and its functional characterization. We also mention that further work to fully reveal this EPS structure will be necessary using higher resolution NMR spectra combined with chemical techniques. This work has given partial structural elucidation, which can be considered a previous background for other research on this topic.	4	1	Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	1
+To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript. In short, they allow to clearly identify the D-mannose α-(1→2) and α-(1→4) linkages in EPS. These results suggest they are the prevailing units of the EPS backbone making it a branched one. Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.	2	1	To my deep regret the structural characteristic of EPS is the weak point of this publication.	1	2	biom12060834_makarova	1
+According to the composition of EPS no metal was found to be attached with the structure of purified EPS. Likewise, in this study the monomeric composition was analysed to demonstrate the structure of purified EPS, where no metal was found to be attached with the chemical structure of it. This is also mentioned and referred in the revised version of the manuscript.	2	1	It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.	1	2	biom12060834_makarova	1
+As suggested by the reviewer, this keyword is removed from the revised version of the manuscript.	2	1	"Line 42: It is premature to include the term ""structure"" in keywords at this stage."	1	2	biom12060834_makarova	1
+Thank you so much dear reviewer for mentioning this po The correction has been made now in the revised version of the manuscript.	2	1	Line 526-527: The proposal should be reformulated. According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.	1	2	biom12060834_makarova	1
+As suggested by the reviewer, the non-pertinent references has been checked throughout the manuscript and unnecessary references are removed too.	2	1	The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.	1	2	biom12060834_perova	1
+As suggested by the reviewer, now the title include the production optimization too and the new title is “Optimization and characterization of a novel exopolysaccharide from Bacillus haynesii CamB6 for food applications”.	2	1	Optimization of production constitutes an important part not mentioned in the title.	1	2	biom12060834_perova	1
+As suggested by the reviewer, the acceptability for food application has now been mentioned with appropriate refences (Nicolaus et al., 2010; Kambourova et al., 2018; Gongi et al., 2022) in the revised version of the manuscript.	2	1	Acceptability for food applications was not discussed.	1	2	biom12060834_perova	1
+According to reviewer´s suggestion, the language of the manuscript is thoroughly revised.	2	1	English language should be revised thoroughly. There are inconsistencies in singular-plural concordance between subject and verb as well as noun and pronoun. Verb tenses should be checked. Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.	1	2	biom12060834_perova	1
+As suggested by the reviewer, the consistent of the word had been checked throughout.	2	1	EPS vs. EPSs in plural form should be consistent throughout.	1	2	biom12060834_perova	1
+Constant abbreviation had been checked throughout in the revised version of the manuscript.	2	1	Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.	1	2	biom12060834_perova	1
+Accoding to the suggestion of the reviewer, Table 1 is revised and both coded – noncoded values are now included.	2	1	l. 160 There is no coded value in Table 1.	1	2	biom12060834_perova	1
+Both the tables (table 1 and table 2) have been placed in the proper place where it expected in the revised manuscript.	2	1	l. 165 Table 2 where Table 1 expected	1	2	biom12060834_perova	1
+The suggested correction has been done in the revised version of the manuscript	2	1	l. 169-170 thirty instead of thirteen ?	1	2	biom12060834_perova	1
+The corrected has been made now.	2	1	l. 177 i instead of 0 in second term?	1	2	biom12060834_perova	1
+The revision has been made as suggested.	2	1	l. 228 GPC defined on l. 231	1	2	biom12060834_perova	1
+as suggested by the reviewer, PEG has been defined in the revised version of the manuscript	2	1	l. 230 PEG undefined	1	2	biom12060834_perova	1
+The volume is now added in the revised version of the manuscript.	2	1	l. 247 Volume of 0.2 mM ethanolic DPPH solution?	1	2	biom12060834_perova	1
+Suggestion has been included in the revised version of the manuscript throughout.	2	1	255, 286, and 441 et al.	1	2	biom12060834_perova	1
+This to kindly mention to the reviewer, the map is the collection site of the EPS producing Bacillus haynesii CamB6. This is a first such report from this collection site, the map is necessary to keep in the manuscript.	2	1	Figure 1 What is the significance of the tick mark labels on the maps? Are these maps really necessary?	1	2	biom12060834_perova	1
+Figure 2 has been modified as suggested by the reviewer.	2	1	Figure 2 No x-axis label Units not specified	1	2	biom12060834_perova	1
+As suggested by the reviewer, Figure 3 is now modified with specified units of the axes and also in the figure legend.	2	1	Figure 3	1	2	biom12060834_perova	1
+All the Suggestion has been included in the figure legend of revised version of the manuscript	2	1	417 and 432 30.0 instead of 3.0 ?	1	2	biom12060834_perova	1
+To avoid the redundancy of the figures as suggested by the reviewer, figure 4 is removed from the revised version of the manuscript.	2	1	Units are not specified on the axes labels or in the legend. Figure 4 This figure is redundant as it gives the same information as Figure 3. Tick label values for glucose and yeast extract are different.	1	2	biom12060834_perova	1
+As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript	2	1	433, 434, 454, 458, and 507 First section 3.6 (and its subsections) should be section 3.5.	1	2	biom12060834_perova	1
+This has been checked now and revised.	2	1	l. 471, 600, 678, and 718 Reference format is different; corresponding references could be missing in the list.	1	2	biom12060834_perova	1
+We performed 2D-NMR analysis , which allows us to determine exactly the α-Manp linkage, which is the major sugar component, and the β- structure of Galp and Glucp.	2	1	l. 480 Why was linkage analysis not performed?	1	2	biom12060834_perova	1
+This is a heteropolysaccharide composed of 3 different sugars as determined for HPLC, which is highly common in this kind of polysaccharides isolated from bacterial strains (10.3390/foods11020156)	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	1
+There are several earlier report where different concentration of yeast extract is added to the culture media for optimized EPS production, and the resultant EPS also concisted of mannan. However, there is no report that the mannan came from yeast extract (DOI: 10.1016/j.ijbiomac.2019.09.139, DOI: 10.1023/B:WIBI.0000033068.45655.2a). The same applies for our study too. In addition, the yeast mannan have β-(1→4) linkage and the one found in this work have α-linkage type. This allows us to conclude they are different polysaccharides.	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	1
+Several 2D-NMR spectra were performed to confirm 1H and 13C chemical shift of EPS structure, and their analysis was added to the manuscript.	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	1
+It was corrected in manuscript	2	1	Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	1
+A new discussion of 1D and 2D NMR spectra was added and corrected.	2	1	l. 500 The chemical shift range is more than from 1 to 5. ll.	1	2	biom12060834_perova	1
+According to the suggestion, section numbering has been checked thoroughly in the revised version of the manuscript.	2	1	l. 537 No section 3.6.2	1	2	biom12060834_perova	1
+As suggested by the reviewer, the necessary revision in done in the manuscript	2	1	l. 636 Glc and Gal	1	2	biom12060834_perova	1
+As suggested by the reviewer, the information is corrected and revised now	2	1	l. 683-684 Food-grade oils are not aliphatic and aromatic hydrocarbons.	1	2	biom12060834_perova	1
+Suggestion has been included in the revised version of the manuscript	2	1	l. 725 possibility of future	1	2	biom12060834_perova	1
+As suggested by the reviewer, minor errors throughout the manuscript has been revised now.	4	1	The English language was improved, but there are still errors.	3	2	biom12060834_perova	1
+Dear reviewer, thanks for your valuable comments for improving the quality of this paper. We will answer in the following paragraphs because all comments were related to the same topic.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+We agree with some of the comments that you made. However, we would like to remind you that the main purpose of this manuscript is not the complete elucidation of the EPS chemical structure but also the optimization of the EPS production, and characterization of the EPS functional properties. At the same time, we provided different chemical properties like: sugar identification and its molar ratio, molecular weight, functional group analysis and thermal stability of the EPS, and this journal has recently published other papers with similar or less chemical information on the EPS, for instance: Kuo, H.-C.; Liu, Y.-W.; Lum, C.-C.; Hsu, K.-D.; Lin, S.-P.; Hsieh, C.-W.; Lin, H.-W.; Lu, T.-Y.; Cheng, K.- C. Ganoderma formosanum Exopolysaccharides Inhibit Tumor Growth via Immunomodulation. Int. J. Mol. Sci. 2021, 22, 11251. https://doi.org/10.3390/ijms222011251 Fetsiukh, A.; Conrad, J.; Bergquist, J.; Timmusk, S. Silica Particles Trigger the Exopolysaccharide Production of Harsh Environment Isolates of Growth-Promoting Rhizobacteria and Increase Their Ability to Enhance Wheat Biomass in Drought-Stressed Soils. Int. J. Mol. Sci. 2021, 22, 6201. https://doi.org/10.3390/ijms22126201.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+On the other side, we performed the 1D and 2D - NMR analysis as complementary information to the other techniques in the present work. Such analysis was performed to try to replace the traditional acid hydrolysis, methylation/acetylation and GC-MS used in glycoside analysis. On the opposite to NMR, we do not have access to this analysis, and we could not find it as a service.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+The HPLC and NMR results from the present work confirmed the heterogeneity of the EPS (3 different sugar moieties with different molar ratios), which agrees with the other reported mannan polysaccharides. The 1H and 13C NMR chemical shifts are comparable with literature reports of other similar EPS or oligosaccharides, where complex structures are identified by NMR too (Casillo et al., 2021; Chatterjee et al; 2018). Besides, our 2D-NMR analysis supported EPS heterogeneity, identifying different types of monomer linkages. Casillo, A., Fabozzi, A., Russo Krauss, I., Parrilli, E., Biggs, C. I., Gibson, M. I., … Corsaro, M. M. (2021). Physicochemical Approach to Understanding the Structure, Conformation, and Activity of Mannan Polysaccharides. Biomacromolecules, 22(4), 1445–1457. doi:10.1021/acs.biomac.0c01659. Chatterjee, S., Mukhopadhyay, S. K., Gauri, S. S., & Dey, S. (2018). Sphingobactan, a new α-mannan exopolysaccharide from Arctic Sphingobacterium sp. IITKGP-BTPF3 capable of biological response modification. International Immunopharmacology, 60, 84–95. doi:10.1016/j.intimp.2018.04.039 •	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+Your mention of the non-uniform peak intensity confirming the three different sugar units found. This is because the signal intensity of NMR peaks is related to an -H abundance. Different monomer ratios are responsible for this variation and support our findings. Also, the 10 H1/H2 cross peaks observed in the COSY spectra supported our findings of EPS heterogeneity, given the following facts: 1) We found 3 different types of sugars by HPLC. 2) We identified by NMR at least 2 types of linkages α 1-2 Man-Man and alfa 1-4 Man-Man monosaccharides. 3) There are other 2 sugars for which the β- type of linkage was identified but not the exact sugar position. 4) Anomeric protons shifts from units at the reducing end of polysaccharide differs from the one in another position. In conclusion, there are different linkage possibilities (plus already identified) to different monosaccharides in different ring positions. This gives rise to a multiplicity of signals (not fully identified in this work), which could produce different H1/H2 correlations.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+About the COSY spectrum discussion, only a minor and partial discussion is presented that can be found below the Scheme in manuscript: “In the COSY spectrum, it was found its α-configuration by low-field H-1 signal at δ 5.34 and correlated with H-2 (δ 4.0), H-6 (δ 3.98 and 3.85), and H-5 (δ 4.12) (Speciale et al., 2022).”	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+Regarding the TOCSY and HMBC mentioned in the methods, they are now deleted from the manuscript, as suggested.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+Finally, we kindly request the reviewer to consider that this is the first approach to studying a Chilean hot spring EPS isolation and its functional characterization. We also mention that further work to fully reveal this EPS structure will be necessary using higher resolution NMR spectra combined with chemical techniques. This work has given partial structural elucidation, which can be considered a previous background for other research on this topic.	4	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	1
+To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript. In short, they allow to clearly identify the D-mannose α-(1→2) and α-(1→4) linkages in EPS. These results suggest they are the prevailing units of the EPS backbone making it a branched one. Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.	2	1	To my deep regret the structural characteristic of EPS is the weak point of this publication. Fourier transform infrared spectroscopy (FTIR) and NMR spectroscopy data can only be regarded as preliminary. According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure. Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc. ), as well as two-dimensional NMR spectroscopy.	1	2	biom12060834_perova	1
+According to the composition of EPS no metal was found to be attached with the structure of purified EPS. Likewise, in this study the monomeric composition was analysed to demonstrate the structure of purified EPS, where no metal was found to be attached with the chemical structure of it. This is also mentioned and referred in the revised version of the manuscript.	2	1	EPC is isolated from a thermal spring of volcanic origin, which contains a set of various elements, including heavy metals. It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.	1	2	biom12060834_perova	1
+As suggested by the reviewer, this keyword is removed from the revised version of the manuscript.	2	1	"Line 42: It is premature to include the term ""structure"" in keywords at this stage."	1	2	biom12060834_perova	1
+Thank you so much dear reviewer for mentioning this point. The correction has been made now in the revised version of the manuscript.	2	1	Line 526-527: The proposal should be reformulated. According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.	1	2	biom12060834_perova	1
+This is not the case; HSA and the other fluids tested as inductors were added to the growth medium (LB) medium, and the cells were allowed to grow in these conditions for 5 h at 37ºC. After that, the cells were collected, and RNA was extracted. Since HSA does not reach the cytosol, at least not in an intact manner, hydrolysis seems not possible during cell growth.	2	1	It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	1
+It is also worth mentioning that the presence of HSA in the growth medium produced a reduction only in some mRNA species. These results speak against an unspecific hydrolyzing effect.	2	1	It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	1
+We also want to underscore that all RNA samples were checked after extraction. Agarose gels electrophoresis confirmed the integrity of RNA and lack of DNA contamination.	2	1	It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	1
+Finally, the article provided by the reviewer describes HSA hydrolysis of extracellular, not intracellular, RNA.	2	1	It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	1
+A follow-up project that will be submitted for publication in the near future extends the studies of the effects produced by human fluids, and their components will include the experiments demonstrating a bactericidal effect of cefiderocol.	2	1	Cefiderocol is a cidal antibiotic. It would be of great interest if for the key results the authors would also provide MBC and / or time-resolved kill-curves. I believe this kind of data would be of great relevance for assessing the clinical relevance of the observed effects.	1	2	biomedicines10030600_makarova	1
+We thank the reviewer for their suggestion and have now computed AMPA-NMDA ratios and included a phrase in lines 424-425 that summarises the statistics for the comparison of AMPA-NMDA ratios between the GluN2B variants (WT and mutants).	2	1	Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative. It follows from the methodology that AMPA currents were recorded on a regular basis.	1	2	brainsci12060789_makarova	1
+We thank the reviewer for this excellent suggestion. We have now measured the decay time constants for the NMDA-EPSCs before and after TCN-201. The graph of the data is shown in a new figure in the supplement, Fig. S4. The finding is insightful, and so we have added a paragraph relating to it in the results section, lines 520-536.	2	1	Please add a chart about tau changes in Figure 3.	1	2	brainsci12060789_makarova	1
+In answer to the last comment here from the reviewer, the time scale in Fig. 3A is correct – the examples chosen are representative of the group data in terms of the amount of block by TCN-201, but these examples all happen to have faster decays than the examples shown in Figures 1 and 2. In any case, to be more consistent with the formatting in some of the other figures, we have lengthened the scale bar and increased the time proportionally.	2	1	Please verify if the time scale is correct in Fig. 3А.	1	2	brainsci12060789_makarova	1
+This is an excellent idea. We have added an additional (final) main text figure, Fig. 7, illustrating how the molecular defects could converge on similar NMDA-EPSCs. We have explained the model in the accompanying figure legend with reference to supporting evidence in other figures.	2	1	A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion	1	2	brainsci12060789_makarova	1
+We have added further discussion relating to the pathogenesis of neurodevelopmental disorders arising from GRIN2B mutations in lines 765-771 of the manuscript.	2	1	A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.	1	2	brainsci12060789_makarova	1
+It is hard to know exactly why the measurements of the internal control, untransfected neuron appear to vary between the experimental groups. While an effort was made to try and evoke synaptic responses with a consistent stimulus voltage setting, the effectiveness of the stimulus for evoking an NMDA-EPSC depends on, among other things, the condition of the stimulating electrode (e.g. resistance, stray capacitance, bubbles etc). It was rarely practical to record all mutant conditions in the same experiment, so variation in the stimulating electrode condition over the course of this series of experiments could potentially lead to some apparent differences between untransfected neurons with respect to their mean NMDA-EPSC peak amplitude and charge transfer. What this does serve to illustrate though is how important it was for us to use untransfected neurons as an internal control.	2	1	Even though fig.2aiii shows a comparable relative peak amplitude, which indicates both GOF and LOF rescue NMDA-EPSC in Grin2b-/- neurons, it looks to me that the absolute amplitude in fig.2aii shows a quite big difference in un-transferred groups which are supposed to be comparable, not as consistent as shown in fig.1ci. How to explain this discrepancy?	1	2	brainsci12060789_makarova	1
+The supplementary figures were within the same PDF so we are not sure why the author had trouble gaining accessing to the figures. Assuming the reviewer had trouble understanding the contents of the supplement, we have tried to simplify somewhat the text in the supplementary figure legends.	2	1	The supplementary figures are inaccessible.	1	2	brainsci12060789_makarova	1
+We thank the reviewer for this comment. Indeed, there were many long sentences and the abstract did not capture all of our findings. We have rewritten the abstract to accommodate the reviewer’s suggestions. The abstract is now 311 words long (almost 40 words shorter than the original abstract), has shorter sentences, and includes some summary of the findings reported in all the main figures.	2	1	The abstract is too long, which does not actually abstract the whole content very well. There are also a number of sentences that are so complicated for easy read. This part should be substantially polished into a shortened and precise style.	1	2	brainsci12060789_makarova	1
+We thank the reviewer for their suggestion and have now computed AMPA-NMDA ratios and included a phrase in lines 424-425 that summarises the statistics for the comparison of AMPA-NMDA ratios between the GluN2B variants (WT and mutants).	2	1	Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative. It follows from the methodology that AMPA currents were recorded on a regular basis.	1	2	brainsci12060789_perova	1
+We thank the reviewer for this excellent suggestion. We have now measured the decay time constants for the NMDA-EPSCs before and after TCN-201. The graph of the data is shown in a new figure in the supplement, Fig. S4. The finding is insightful, and so we have added a paragraph relating to it in the results section, lines 520-536. In answer to the last comment here from the reviewer, the time scale in Fig. 3A is correct – the examples chosen are representative of the group data in terms of the amount of block by TCN-201, but these examples all happen to have faster decays than the examples shown in Figures 1 and 2. In any case, to be more consistent with the formatting in some of the other figures, we have lengthened the scale bar and increased the time proportionally.	2	1	Please add a chart about tau changes in Figure 3. Please verify if the time scale is correct in Fig. 3А.	1	2	brainsci12060789_perova	1
+This is an excellent idea. We have added an additional (final) main text figure, Fig. 7, illustrating how the molecular defects could converge on similar NMDA-EPSCs. We have explained the model in the accompanying figure legend with reference to supporting evidence in other figures.	2	1	A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion	1	2	brainsci12060789_perova	1
+We have added further discussion relating to the pathogenesis of neurodevelopmental disorders arising from GRIN2B mutations in lines 765-771 of the manuscript.	2	1	A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.	1	2	brainsci12060789_perova	1
+It is hard to know exactly why the measurements of the internal control, untransfected neuron appear to vary between the experimental groups. While an effort was made to try and evoke synaptic responses with a consistent stimulus voltage setting, the effectiveness of the stimulus for evoking an NMDA-EPSC depends on, among other things, the condition of the stimulating electrode (e.g. resistance, stray capacitance, bubbles etc). It was rarely practical to record all mutant conditions in the same experiment, so variation in the stimulating electrode condition over the course of this series of experiments could potentially lead to some apparent differences between untransfected neurons with respect to their mean NMDA-EPSC peak amplitude and charge transfer. What this does serve to illustrate though is how important it was for us to use untransfected neurons as an internal control.	2	1	Even though fig.2aiii shows a comparable relative peak amplitude, which indicates both GOF and LOF rescue NMDA-EPSC in Grin2b-/- neurons, it looks to me that the absolute amplitude in fig.2aii shows a quite big difference in un-transferred groups which are supposed to be comparable, not as consistent as shown in fig.1ci. How to explain this discrepancy?	1	2	brainsci12060789_perova	1
+The supplementary figures were within the same PDF so we are not sure why the author had trouble gaining accessing to the figures. Assuming the reviewer had trouble understanding the contents of the supplement, we have tried to simplify somewhat the text in the supplementary figure legends.	2	1	The supplementary figures are inaccessible.	1	2	brainsci12060789_perova	1
+We thank the reviewer for this comment. Indeed, there were many long sentences and the abstract did not capture all of our findings. We have rewritten the abstract to accommodate the reviewer’s suggestions. The abstract is now 311 words long (almost 40 words shorter than the original abstract), has shorter sentences, and includes some summary of the findings reported in all the main figures. The abstract now reads as follows: GRIN2B mutations are rare but often associated with patients having severe neurodevelopmental disorders, including a varying range of symptoms such as intellectual disability, developmental delay and epilepsy. Patient symptoms likely arise from mutations disturbing the role that the encoded NMDA receptor subunit, GluN2B, plays at neuronal connections in the developing nervous system. In this study, we have investigated the cell-autonomous effects of putative gain- (GoF) and loss-of-function (LoF) missense GRIN2B mutations on excitatory synapses onto CA1 pyramidal neurons in organotypic hippocampal slices. In the absence of both native GluN2A and GluN2B subunits, functional incorporation into synaptic NMDA receptors was attenuated for GoF mutants, or almost eliminated for LoF GluN2B mutants. NMDA receptor-mediated excitatory postsynaptic currents (NMDA-EPSCs) from synaptic GoF GluN1/2B receptors had prolonged decays consistent with their functional classification. Nonetheless, in the presence of native GluN2A, molecular replacement of native GluN2B with GoF and LoF GluN2B mutants all led to similar functional incorporation into synaptic receptors, more rapidly decaying NMDA-EPSCs and greater inhibition by TCN-201, a selective antagonist of GluN2A-containing NMDA receptors. Mechanistic insight was gained from experiments in HEK293T cells, which revealed that GluN2B GoF mutants slowed deactivation in diheteromeric GluN1/2B, but not triheteromeric GluN1/2A/2B receptors. We also show that a disease-associated missense mutation, which severely affects surface expression, causes opposing effects on NMDA-EPSC decay and charge transfer when introduced into GluN2A or GluN2B. Finally, we show that having a single null Grin2b allele has only a modest effect on NMDA-EPSC decay kinetics. Our results demonstrate that functional incorporation of GoF and LoF GluN2B mutants into synaptic receptors and the effects on EPSC decay times are highly dependent on the presence of triheteromeric GluN1/2A/2B NMDA receptors, thereby influencing the functional classification of NMDA receptor variants as GoF or LoF mutations. These findings highlight the complexity of interpreting effects of disease-causing NMDA receptor missense mutations in the context of neuronal function.	2	1	The abstract is too long, which does not actually abstract the whole content very well. There are also a number of sentences that are so complicated for easy read. This part should be substantially polished into a shortened and precise style.	1	2	brainsci12060789_perova	1
+We concur that a brief description of the literature regarding EIH in FM is warranted and have added information to the discussion as follows: “Previous research regarding the effects of acute bouts of exercise on pain sensitivity in FM patients is largely equivocal. Some studies have demonstrated a hypoalgesic effect [5,24] while other studies show either no changes in pain perception or an exacerbation of pain [25–27].”	2	1	However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls.	1	2	brainsci6010008_makarova	1
+With respect to the reviewer’s comment regarding the lack of EIH in controls, we have included some additional information to demonstrate the consistency of our findings with previous research. The passage now reads: “However, it appears that the exercise bout as prescribed was not a sufficient stimulus to induce a decrease in pain sensitivity in our pain-free controls. This is consistent with previous literature regarding the effects of acute exercise on pain in healthy controls, which typically finds that higher intensity exercise is necessary to elicit a hypoalgesic response [33].” Additionally, we want to again mention that our study was not primarily designed to induce EIH in controls; it was designed to examine brain responses to pain following exercise and thus we wanted to ensure that FM patients could complete the exercise bout. Exploring brain responses associated with EIH in healthy individuals would be an interesting future direction.	2	1	The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_makarova	1
+With respect to the reviewer’s comment on generalizability, we agree that our exclusionary criteria were stringent and have added the following sentence to our limitations section. “Further, we excluded individuals with diagnosed mental health conditions and those who were taking medications that could impact pain or the interpretation of brain responses. As such our results may not apply to FM patients with comorbid conditions.”	2	1	Also, the discussion section should be revised to acknowledge that many patients with FM would have been excluded from participation in this study due to major depression and medication consumption so there may be concerns about the generalizability of the sample, BUT the authors appropriately compared the Fibromyalgia Impact Questionnaire scores of their sample to normative data and found no significant difference.	1	2	brainsci6010008_makarova	1
+Lastly, with respect to the comment about controlling for physiological noise, the following statements have been added. “Though the evidence is equivocal, it has been suggested that cardiovascular mechanisms may be involved in the hypoalgesic response to exercise [34] and thus statistically controlling for these effects may have influenced the interpretation of our results. However, without controlling physiological differences between EX and QR the neuroimaging data would have been very difficult to interpret.” I don’t see that the reference actually indicates studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	2	1	In addition, the authors might want to consider and mention that their method of controlling “physiological noise” might have influenced the results because cardiovascular reactions to exercise were, at one time, proposed to be a mechanism of EIH.	1	2	brainsci6010008_makarova	1
+This sentence has been revised to better reflect the reference used.	2	1	P. 1, L. 40 – I don’t see that the reference actually includes studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	1	2	brainsci6010008_makarova	1
+We agree. We consulted with each physician with respect to type and dosage of medication, including whether this was a low, moderate or high dose. We also double-checked each medication and dose with the Physician’s Desk Reference (60th-65th editions). We have added the following information to the methods section “Medication information and dosage were supplied by the patient and their physician and dosage levels (low, moderate, high) were determined through both physician consultation and use of the Physician’s Desk Reference (65th edition).	2	1	P. 2, L. 18 – A reference for the determination that a dosage of antidepressants was “high” should be provided.	1	2	brainsci6010008_makarova	1
+This addition has been made.	2	1	P. 3, L. 15 – The time post-exercise before scanning was provided, but not the time post-quiet rest.	1	2	brainsci6010008_makarova	1
+The suggested change has been made.	2	1	P. 6, L. 5 – “Elevations” should be revised to “higher” so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.	1	2	brainsci6010008_makarova	1
+Thank you for catching this omission. This was an oversight on our part and these effect sizes are now included on page 11.	2	1	P. 6, L. 12 - I am uncertain why the authors only report the effect sizes of group differences for the first run.s	1	2	brainsci6010008_makarova	1
+These analyses were intended primarily for descriptive purposes. However, we recognize the need to control for multiple comparisons in order to reduce the risk of making Type I errors. Therefore, we created 3 families including 2 correlations each (one for each of the significant regions) and performed a Bonferroni correction, making the critical alpha level for significance 0.025. This has been clarified in the statistical analysis section.	2	1	However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_makarova	1
+This change has been made.	2	1	Nine individuals in each group were included in neuroimaging analyses, this should be indicated in the abstract.	1	2	brainsci6010008_makarova	1
+This correction has been made.	2	1	"In table 3 the subheading ""Peak X, Y, X"" needs correction."	1	2	brainsci6010008_makarova	1
+We agree that this would add valuable information to the discussion section and have added the paragraph shown below discussing previous work using neuroimaging to understand the effects of exercise on the brain.	2	1	Consider discussing the results in relation to previous studies on exercise and neuroimaging	1	2	brainsci6010008_makarova	1
+A growing number of studies have begun to employ neuroimaging methods to better understand the impact of exercise on the brain both longitudinally and acutely. For example, Smith and colleagues [37] conducted fMRI scans pre and post an exercise training program in older adults with mild cognitive impairment and found that exercise improved neural efficiency during cognitive tasks post-intervention. Structural MRI has also been used to show the neuroprotective effects of regular exercise in older adults with respect to preservation of brain volume[38]. In contrast to using neuroimaging to track changes in the brain over time, neuroimaging during and immediately following exercise presents some unique challenges due to artifacts associated with movement and the physiological underpinnings of many neuroimaging methods (e.g. BOLD response). EEG has been used most extensively to explore the effects of exercise on cortical activity [39]. PET and fMRI have also been used, though to a much lesser extent. For example, Boecker and colleagues used PET to demonstrate the effects of a long-distance run on opioid release in the brain and Janse Van Rensberg and colleagues used fMRI to examine brain responses to nicotine craving following 10 minutes of moderate intensity cycling. Our study adds to this important body of literature by using fMRI to show that an acute bout of moderate intensity exercise improved brain mechanisms underlying pain modulation in patients with chronic pain and further highlights the potential benefits of utilizing neuroimaging technology to better understand the more immediate effects of exercise on the human brain.	2	1	The findings are novel and may be compared to previous studies of exercise and neuroimaging in fibromyalgia.	1	2	brainsci6010008_makarova	1
+We concur that a brief description of the literature regarding EIH in FM is warranted and have added information to the discussion as follows: “Previous research regarding the effects of acute bouts of exercise on pain sensitivity in FM patients is largely equivocal. Some studies have demonstrated a hypoalgesic effect [5,24] while other studies show either no changes in pain perception or an exacerbation of pain [25–27].”	2	1	However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls. The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_perova	1
+With respect to the reviewer’s comment regarding the lack of EIH in controls, we have included some additional information to demonstrate the consistency of our findings with previous research. The passage now reads: “However, it appears that the exercise bout as prescribed was not a sufficient stimulus to induce a decrease in pain sensitivity in our pain-free controls. This is consistent with previous literature regarding the effects of acute exercise on pain in healthy controls, which typically finds that higher intensity exercise is necessary to elicit a hypoalgesic response [33].” Additionally, we want to again mention that our study was not primarily designed to induce EIH in controls; it was designed to examine brain responses to pain following exercise and thus we wanted to ensure that FM patients could complete the exercise bout. Exploring brain responses associated with EIH in healthy individuals would be an interesting future direction.	2	1	However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls. The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_perova	1
+With respect to the reviewer’s comment on generalizability, we agree that our exclusionary criteria were stringent and have added the following sentence to our limitations section. “Further, we excluded individuals with diagnosed mental health conditions and those who were taking medications that could impact pain or the interpretation of brain responses. As such our results may not apply to FM patients with comorbid conditions.”	2	1	Also, the discussion section should be revised to acknowledge that many patients with FM would have been excluded from participation in this study due to major depression and medication consumption so there may be concerns about the generalizability of the sample, BUT the authors appropriately compared the Fibromyalgia Impact Questionnaire scores of their sample to normative data and found no significant difference.	1	2	brainsci6010008_perova	1
+Lastly, with respect to the comment about controlling for physiological noise, the following statements have been added. “Though the evidence is equivocal, it has been suggested that cardiovascular mechanisms may be involved in the hypoalgesic response to exercise [34] and thus statistically controlling for these effects may have influenced the interpretation of our results. However, without controlling physiological differences between EX and QR the neuroimaging data would have been very difficult to interpret.” I don’t see that the reference actually indicates studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	2	1	In addition, the authors might want to consider and mention that their method of controlling “physiological noise” might have influenced the results because cardiovascular reactions to exercise were, at one time, proposed to be a mechanism of EIH.	1	2	brainsci6010008_perova	1
+This sentence has been revised to better reflect the reference used.	2	1	P. 1, L. 40 – I don’t see that the reference actually includes studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	1	2	brainsci6010008_perova	1
+We agree. We consulted with each physician with respect to type and dosage of medication, including whether this was a low, moderate or high dose. We also double-checked each medication and dose with the Physician’s Desk Reference (60th-65th editions). We have added the following information to the methods section “Medication information and dosage were supplied by the patient and their physician and dosage levels (low, moderate, high) were determined through both physician consultation and use of the Physician’s Desk Reference (65th edition).	2	1	P. 2, L. 18 – A reference for the determination that a dosage of antidepressants was “high” should be provided.	1	2	brainsci6010008_perova	1
+This addition has been made.	2	1	P. 3, L. 15 – The time post-exercise before scanning was provided, but not the time post-quiet rest.	1	2	brainsci6010008_perova	1
+The suggested change has been made.	2	1	P. 6, L. 5 – “Elevations” should be revised to “higher” so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.	1	2	brainsci6010008_perova	1
+Thank you for catching this omission. This was an oversight on our part and these effect sizes are now included on page 11.	2	1	P. 6, L. 12 - I am uncertain why the authors only report the effect sizes of group differences for the first run.s	1	2	brainsci6010008_perova	1
+These analyses were intended primarily for descriptive purposes. However, we recognize the need to control for multiple comparisons in order to reduce the risk of making Type I errors. Therefore, we created 3 families including 2 correlations each (one for each of the significant regions) and performed a Bonferroni correction, making the critical alpha level for significance 0.025.	2	1	However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_perova	1
+This has been clarified in the statistical analysis section.	2	1	However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_perova	1
+This change has been made.	2	1	Nine individuals in each group were included in neuroimaging analyses, this should be indicated in the abstract.	1	2	brainsci6010008_perova	1
+This correction has been made.	2	1	"In table 3 the subheading ""Peak X, Y, X"" needs correction."	1	2	brainsci6010008_perova	1
+We agree that this would add valuable information to the discussion section and have added the paragraph shown below discussing previous work using neuroimaging to understand the effects of exercise on the brain.	2	1	Consider discussing the results in relation to previous studies on exercise and neuroimaging	1	2	brainsci6010008_perova	1
+A growing number of studies have begun to employ neuroimaging methods to better understand the impact of exercise on the brain both longitudinally and acutely. For example, Smith and colleagues [37] conducted fMRI scans pre and post an exercise training program in older adults with mild cognitive impairment and found that exercise improved neural efficiency during cognitive tasks post-intervention. Structural MRI has also been used to show the neuroprotective effects of regular exercise in older adults with respect to preservation of brain volume[38]. In contrast to using neuroimaging to track changes in the brain over time, neuroimaging during and immediately following exercise presents some unique challenges due to artifacts associated with movement and the physiological underpinnings of many neuroimaging methods (e.g. BOLD response). EEG has been used most extensively to explore the effects of exercise on cortical activity [39]. PET and fMRI have also been used, though to a much lesser extent. For example, Boecker and colleagues used PET to demonstrate the effects of a long-distance run on opioid release in the brain and Janse Van Rensberg and colleagues used fMRI to examine brain responses to nicotine craving following 10 minutes of moderate intensity cycling. Our study adds to this important body of literature by using fMRI to show that an acute bout of moderate intensity exercise improved brain mechanisms underlying pain modulation in patients with chronic pain and further highlights the potential benefits of utilizing neuroimaging technology to better understand the more immediate effects of exercise on the human brain.	2	1	The results are clearly reported and adequately discussed. The findings are novel and may be compared to previous studies of exercise and neuroimaging in fibromyalgia.	1	2	brainsci6010008_perova	1
+In the revised manuscript, this was corrected as suggested.	2	1	"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."	1	2	brainsci6010009_makarova	1
+Both cases and controls could have been diagnosed with autism, but it should have been a relative minority. Autism is relatively rare compared to a 314 diagnosis. This issue was added to the study limitations in the revised manuscript. “In the present study, neurodevelopmental diagnoses other than 314.xx were not examined among cases and controls. This limitation of the present study should have had a limited impact on the results observed because of the rarity of other neurodevelopmental diagnoses as compared to a 314.xx diagnosis, but future studies could further evaluate this phenomenon.” Comment 3.	2	1	In the exposed and control groups were there other related diseases such as Autism? If so, how many and what would happen to the analysis if you removed them?	1	2	brainsci6010009_makarova	1
+We don’t have the exact number, but it is assumed to be a relatively small number.	2	1	On page 3 line 22, they describe children with the diagnosis before the exposure. How many were there that fit in this category?	1	2	brainsci6010009_makarova	1
+In the revised manuscript, this was corrected with a better reference.	2	1	On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point. Are there better references to substantiate this point?	1	2	brainsci6010009_makarova	1
+In the revised manuscript, this information was added. “This database is available to outside researchers after obtaining approval from the CDC and KP. Information regarding access to this database is at: http://www.cdc.gov/vaccinesafety/ensuringsafety/monitoring/vsd/accessing-data.html.”	2	1	Can the authors address this point: is this database publicly available?	1	2	brainsci6010009_makarova	1
+Thank you.	2	1	The results are controversial. However, the findings may be of great interest for the readers of the journal. The paper is of high quality. I recommend the manuscript published. It can be published as it is.	1	2	brainsci6010009_makarova	1
+In the revised manuscript, this was corrected as suggested.	2	1	"1. In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."	1	2	brainsci6010009_perova	1
+Both cases and controls could have been diagnosed with autism, but it should have been a relative minority. Autism is relatively rare compared to a 314 diagnosis. This issue was added to the study limitations in the revised manuscript. “In the present study, neurodevelopmental diagnoses other than 314.xx were not examined among cases and controls. This limitation of the present study should have had a limited impact on the results observed because of the rarity of other neurodevelopmental diagnoses as compared to a 314.xx diagnosis, but future studies could further evaluate this phenomenon.”	2	1	2. In the exposed and control groups were there other related diseases such as Autism? If so, how many and what would happen to the analysis if you removed them?	1	2	brainsci6010009_perova	1
+We don’t have the exact number, but it is assumed to be a relatively small number.	2	1	3. On page 3 line 22, they describe children with the diagnosis before the exposure. How many were there that fit in this category?	1	2	brainsci6010009_perova	1
+In the revised manuscript, this was corrected with a better reference.	2	1	On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point. Are there better references to substantiate this point?	1	2	brainsci6010009_perova	1
+In the revised manuscript, this information was added. “This database is available to outside researchers after obtaining approval from the CDC and KP. Information regarding access to this database is at: http://www.cdc.gov/vaccinesafety/ensuringsafety/monitoring/vsd/accessing-data.html.”	2	1	5. Can the authors address this point: is this database publicly available?	1	2	brainsci6010009_perova	1
+Thank you.	2	1	This is an important study that deserves to be published. The results are controversial. However, the findings may be of great interest for the readers of the journal. The paper is of high quality. I recommend the manuscript published. It can be published as it is.	1	2	brainsci6010009_perova	1
+Thank you for the enthusiastic comments.	2	1	This is a well-written paper with a large sample of girls that replicates and extends important research on relational aggression. The focus on the functions (reactive and proactive) of relational aggression has significant implications for intervention with adolescent girls. I commend the authors for this timely and crucial study on factors that influence the development of aggression and antisocial behavior in girls, which is typically an understudied topic.	1	2	bs5040518_makarova	1
+This is now provided in the results section.	2	1	First, it is recommended that the authors provide more information about the selection of the cluster solution. That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth. It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.	1	2	bs5040518_makarova	1
+Thank you! I have now carefully read and edited the paper.	2	1	Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.	1	2	bs5040518_makarova	1
+True that we do not know the direction of effects. Here, we use predictor in the statistical sense not causal sense.	2	1	First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue.	1	2	bs5040518_makarova	1
+We agree this would be useful but it was not our aim to look at interactions. We also believe interactions such as suggested are better done in longitudinal studies where one can examine moderators.	2	1	Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_makarova	1
+We have attempted to be mindful of causal language and only use it where prior research indicates a direction.	2	1	Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.	1	2	bs5040518_makarova	1
+We agree this is a major flaw and we thank the reviewer for pointing out projection bias. We now include this in the limitations in the discussion.	2	1	Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.	1	2	bs5040518_makarova	1
+We agree with the reviewer that there is no easy answer to this problem. We now include a discussion of this in the introduction and cite Lynam and colleagues on this exact dilemma. In their paper, they discuss the Perils of Partialing (p.4, line 15), which is what you do when you control for the overlap. That is, one may be removing the reliable aspect of the self-report measures of aggression (when they are highly correlated) and remaining with residual error.	2	1	A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).	1	2	bs5040518_makarova	1
+The reactive aggressive group did not differ from the combined group on peer delinquency. Thus, both were high on delinquency and peer delinquency. Also, we comment on the overlap between peer delinquency and self-report of delinquency in the results section (p.9, line 14).	2	1	Some findings are difficult to reconcile with the current literature; in addition, they are internally inconsistent. For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?	1	2	bs5040518_makarova	1
+We have carefully edited the paper.	2	1	There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.	1	2	bs5040518_makarova	1
+The manuscript has been edited for grammar problems and for clarity. We have also added Table 1 to assist with understanding our predictors and the broader factors they represent in this study.	2	1	There are enough variables that I hard a hard time following the analyses. A tighter focus in the intro and matching of analyses to hypotheses may help here.	1	2	bs5040518_makarova	1
+Thank you for the enthusiastic comments.	2	1	This is a well-written paper with a large sample of girls that replicates and extends important research on relational aggression. The focus on the functions (reactive and proactive) of relational aggression has significant implications for intervention with adolescent girls. I commend the authors for this timely and crucial study on factors that influence the development of aggression and antisocial behavior in girls, which is typically an understudied topic.	1	2	bs5040518_perova	1
+This is now provided in the results section.	2	1	First, it is recommended that the authors provide more information about the selection of the cluster solution. That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth. It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.	1	2	bs5040518_perova	1
+Thank you! I have now carefully read and edited the paper.	2	1	Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.	1	2	bs5040518_perova	1
+True that we do not know the direction of effects. Here, we use predictor in the statistical sense not causal sense.	2	1	Although the rationale for selecting each of the 9 factors that might be differentially related to reactive or proactive relational aggression is convincing, the overall picture is incomplete on at least two accounts. First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue. Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_perova	1
+We agree this would be useful but it was not our aim to look at interactions. We also believe interactions such as suggested are better done in longitudinal studies where one can examine moderators.	2	1	Although the rationale for selecting each of the 9 factors that might be differentially related to reactive or proactive relational aggression is convincing, the overall picture is incomplete on at least two accounts. First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue. Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_perova	1
+We have attempted to be mindful of causal language and only use it where prior research indicates a direction.	2	1	The use of a cross-sectional design is a major limitation, because it cannot help determine the directionality of the links between the nine factors and subtypes of relational aggression, left alone causality. This bears directly on the issue of predictors vs. correlates vs. consequences of subtypes of relational aggression. Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.	1	2	bs5040518_perova	1
+We agree this is a major flaw and we thank the reviewer for pointing out projection bias. We now include this in the limitations in the discussion.	2	1	All the measures are self-reported. This artificially inflates the link between the study variables. Acknowledging this problem in the limitations does not solve it. Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.	1	2	bs5040518_perova	1
+We agree with the reviewer that there is no easy answer to this problem. We now include a discussion of this in the introduction and cite Lynam and colleagues on this exact dilemma. In their paper, they discuss the Perils of Partialing (p.4, line 15), which is what you do when you control for the overlap. That is, one may be removing the reliable aspect of the self-report measures of aggression (when they are highly correlated) and remaining with residual error.	2	1	Creating groups based on a cluster analysis may have created unnecessary problems. In particular, and contrary to what the authors seem to believe, it is not possible to know whether differences between the two aggressive groups reflect differences in levels of relational aggression or in type of relational aggression (reactive only vs. combined), given that the combined group is obviously much more aggressive than the reactive-only group. A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).	1	2	bs5040518_perova	1
+The reactive aggressive group did not differ from the combined group on peer delinquency. Thus, both were high on delinquency and peer delinquency. Also, we comment on the overlap between peer delinquency and self-report of delinquency in the results section (p.9, line 14).	2	1	Some findings are difficult to reconcile with the current literature; in addition, they are internally inconsistent. For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?	1	2	bs5040518_perova	1
+We have carefully edited the paper.	2	1	There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.	1	2	bs5040518_perova	1
+The manuscript has been edited for grammar problems and for clarity.	2	1	There are enough variables that I hard a hard time following the analyses. A tighter focus in the intro and matching of analyses to hypotheses may help here.	1	2	bs5040518_perova	1
+We have also added Table 1 to assist with understanding our predictors and the broader factors they represent in this study.	2	1	As it is, this reviewer had difficulty following the variables as predictors/outcomes and the hypotheses that are driving the paper 2.	1	2	bs5040518_perova	1
+Thank you for this advice. We have trimmed the paper to focus on two of the five elements of sexual subjectivity - only those that were expected to be most relevant to partnered sexual partner: entitlement to sexual partner pleasure and efficacy in achieving pleasure. This has substantially limited the analyses conducted. We provide a rationale for this (see the first paragraph on p.4) and have thoroughly revised the entire paper.	2	1	With better theoretical framing and a more limited set of analyses, the value of the study results should be more clear.	1	2	bs6010004_makarova	1
+We have done this and have also substantially revised the entire Introduction (see new section starting on p. 3 titled Definition and measurement).	2	1	Also, a more thorough description of the measurement of the construct and the validity of the measurement could be described in that section.	1	2	bs6010004_makarova	1
+We have corrected this sentence.	2	1	On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” **	1	2	bs6010004_makarova	1
+We have added to our theoretical explanation for why there would be gender differences (see the section titled Gender Differences in Sexual Behavior, Attitudes, and Sexual Subjectivity, p. 4).	2	1	The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.	1	2	bs6010004_makarova	1
+We have substantially revised all sections of this paper to improve the flow and removed some headings.	2	1	The authors rely too much on headings to transition between ideas. The paper would flow more smoothly if some transition sentences were included.	1	2	bs6010004_makarova	1
+We have also added hypotheses (see p. 5-6)	2	1	More specific hypotheses could be proposed.	1	2	bs6010004_makarova	1
+We have revised this section (The Current Study, p.5).	2	1	The section where research questions are described is worded in an awkward way.	1	2	bs6010004_makarova	1
+We have moved this content on group difference to the first section of the Results section.	2	1	The reporting of group differences in the participants section might be better placed in the results section after the measures are described.	1	2	bs6010004_makarova	1
+Given our focus on only two element of sexual subjectivity, this was no longer an issue because items were the same on measures for males and females.	2	1	The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained. Perhaps factor analysis could be used to determine appropriate subscales?	1	2	bs6010004_makarova	1
+"We have added this information on p. 7 in the Procedure section. It reads: Approval from the university Human Research Ethics Committee was obtained prior to data collection. At Time 1 (T1), participants were approached at a university campus in Australia in the week before classes commenced (i.e., during orientation week) and asked to participate in a study ""About You and Your Relationships."" The front cover of the survey described the questions as focused on personal sexual and romantic experiences, and stressed the confidential nature of the survey."	2	1	What were participants told about the purpose of the study?	1	2	bs6010004_makarova	1
+Reducing the number of elements of sexual subjectivity has reduced the number of tests performed.	2	1	Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.	1	2	bs6010004_makarova	1
+We have added hypotheses to the section titled The Current Study on p. 5-6.	2	1	Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).	1	2	bs6010004_makarova	1
+We have now used multiple imputation to maintain all 295 participants in all analyses.	2	1	Analyses should be re-done using a more sophisticated method for handling missing data, such as multiple imputation or full-information maximum likelihood (Shafer & Graham, 2002, Psychological Methods).	1	2	bs6010004_makarova	1
+We are sorry that we did not keep a tally of the number of students approached. We have added to the Limitations section of the Discussion about this (see p. 12). It reads, First, the participants in the study were predominantly middle class, Caucasian university students, residing in one region of Australia. There is also the possibility that individuals more interested in sex and relationships chose to participate. Therefore, the results of the current study may have some limited generalizability.	2	1	At the very least, the Discussion should consider how generalizable these results are.	1	2	bs6010004_makarova	1
+We agree that this is very important. We have added the Introduction to expand on these points. Unfortunately we could not conduct these suggested analyses. We had very little change in sexual behavior given the way we measured it. Also, we did not have confidence in estimation of sexual behavior for those who did not participate at Wave 2, which could have increased the change. Thus, we have maintained our original analyses, but make this comment in the Discussion (see p. 12-13): Second, probably because the majority of the participants had already experienced coitus prior to the first data collection and because we measured types of sexual behavior and not frequency or some other aspect of behavior, there was high stability in sexual behavior over the one year of this study. Recent statistics show that the majority of Australian year 10-12 students (approximately 15-17 years) have engaged in some form of sexual behavior [12]. Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with change in sexual behavior, future studies may need to begin with a younger population. Recently, Hensel et al. [31] found that, in their adolescent participants, sexual self-concept had a bidirectional relationship with sexual experience, and the same may occur for sexual subjectivity. Future research could examine the possibility of bidirectional relationships between sexual subjectivity and sexual behavior. In addition, no previous study has examined the timing of pubertal development and sexual subjectivity. Evidence suggests that adolescents that mature earlier, compared to their peers, form romantic relationships earlier and experience sexual behaviors earlier [11,55]. In the current and past research, the link between age and sexual subjectivity has been relatively weak [34]. Substituting timing of pubertal status for age may provide more evidence regarding individual characteristics that influence differences in sexual subjectivity and its development over time.	2	1	More generally, I think the putative timescale of effects of behavior on sexual subjectivity needs more theoretical elaboration. Are behaviors in early adolescence (before age 16), for example, expected to continue to influence the trajectory of sexual subjectivity regardless of later sexual behavior?	1	2	bs6010004_makarova	1
+We have trimmed the paper to focus on two of the five elements of sexual subjectivity and anticipated that these two elements would show different results. Thus, we have maintained the separate analyses of these two elements. Please see the added Hypotheses on p. 5-6.	2	1	The shift to a latent factor approach would have the added benefit of allowing the authors to use FIML to account for missing data at T2, and they could thus use their entire sample.	1	2	bs6010004_makarova	1
+We have removed these analyses from the paper in an attempt to reduce the number of analyses and streamline the entire paper.	2	1	This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.	1	2	bs6010004_makarova	1
+Thank you for this suggestion. The reference has been added to the paper.	2	1	I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion.	1	2	bs6010004_makarova	1
+Thank you for this advice. We have trimmed the paper to focus on two of the five elements of sexual subjectivity - only those that were expected to be most relevant to partnered sexual partner: entitlement to sexual partner pleasure and efficacy in achieving pleasure. This has substantially limited the analyses conducted. We provide a rationale for this (see the first paragraph on p.4) and have thoroughly revised the entire paper.	2	1	my main concern with the study is that there were too many analyses presented to interpret in a meaningful way. The authors should revise the paper providing more theoretical background and accompanying analyses. With better theoretical framing and a more limited set of analyses, the value of the study results should be more clear.	1	2	bs6010004_perova	1
+We have done this and have also substantially revised the entire Introduction (see new section starting on p. 3 titled Definition and measurement).	2	1	I think it would be preferable to describe the measurement of sexual subjectivity shortly after it is defined on p. 3. Also, a more thorough description of the measurement of the construct and the validity of the measurement could be described in that section.	1	2	bs6010004_perova	1
+We have corrected this sentence.	2	1	On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” **	1	2	bs6010004_perova	1
+We have added to our theoretical explanation for why there would be gender differences (see the section titled Gender Differences in Sexual Behavior, Attitudes, and Sexual Subjectivity, p. 4).	2	1	The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.	1	2	bs6010004_perova	1
+We have substantially revised all sections of this paper to improve the flow and removed some headings.	2	1	The authors rely too much on headings to transition between ideas. The paper would flow more smoothly if some transition sentences were included.	1	2	bs6010004_perova	1
+We have revised this section (The Current Study, p.5). We have also added hypotheses (see p. 5-6)	2	1	More specific hypotheses could be proposed. The section where research questions are described is worded in an awkward way.	1	2	bs6010004_perova	1
+We have moved this content on group difference to the first section of the Results section.	2	1	The reporting of group differences in the participants section might be better placed in the results section after the measures are described.	1	2	bs6010004_perova	1
+Given our focus on only two element of sexual subjectivity, this was no longer an issue because items were the same on measures for males and females.	2	1	The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained. Perhaps factor analysis could be used to determine appropriate subscales?	1	2	bs6010004_perova	1
+"We have added this information on p. 7 in the Procedure section. It reads: Approval from the university Human Research Ethics Committee was obtained prior to data collection. At Time 1 (T1), participants were approached at a university campus in Australia in the week before classes commenced (i.e., during orientation week) and asked to participate in a study ""About You and Your Relationships."" The front cover of the survey described the questions as focused on personal sexual and romantic experiences, and stressed the confidential nature of the survey."	2	1	What were participants told about the purpose of the study?	1	2	bs6010004_perova	1
+Reducing the number of elements of sexual subjectivity has reduced the number of tests performed.	2	1	There is an increase in Type I error with the multiple t-tests conducted thatshould be addressed. Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.	1	2	bs6010004_perova	1
+We have added hypotheses to the section titled The Current Study on p. 5-6.	2	1	Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).	1	2	bs6010004_perova	1
+We have now used multiple imputation to maintain all 295 participants in all analyses.	2	1	First, the attrition between Time 1 and Time 2 was substantial (40% of participants), and this is handled using list-wise deletion. That is, any one who did not complete both assessments was not included in the analyses. List-wise deletion can lead to serious bias in results. Analyses should be re-done using a more sophisticated method for handling missing data, such as multiple imputation or full-information maximum likelihood (Shafer & Graham, 2002, Psychological Methods).	1	2	bs6010004_perova	1
+We are sorry that we did not keep a tally of the number of students approached. We have added to the Limitations section of the Discussion about this (see p. 12). It reads, First, the participants in the study were predominantly middle class, Caucasian university students, residing in one region of Australia. There is also the possibility that individuals more interested in sex and relationships chose to participate. Therefore, the results of the current study may have some limited generalizability.	2	1	"Second, potential participants were ""approached at a university campus"" during orientation week. What percentage of people who were approached agreed to participate? How broadly representative of the university population (or the university-age population) is the sample? From the relatively high percentage of non-heterosexual participants, there seems to be some selection bias. At the very least, the Discussion should consider how generalizable these results are."	1	2	bs6010004_perova	1
+We agree that this is very important. We have added the Introduction to expand on these points. Unfortunately we could not conduct these suggested analyses. We had very little change in sexual behavior given the way we measured it. Also, we did not have confidence in estimation of sexual behavior for those who did not participate at Wave 2, which could have increased the change. Thus, we have maintained our original analyses, but make this comment in the Discussion (see p. 12-13): Second, probably because the majority of the participants had already experienced coitus prior to the first data collection and because we measured types of sexual behavior and not frequency or some other aspect of behavior, there was high stability in sexual behavior over the one year of this study. Recent statistics show that the majority of Australian year 10-12 students (approximately 15-17 years) have engaged in some form of sexual behavior [12]. Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with change in sexual behavior, future studies may need to begin with a younger population. Recently, Hensel et al. [31] found that, in their adolescent participants, sexual self-concept had a bidirectional relationship with sexual experience, and the same may occur for sexual subjectivity. Future research could examine the possibility of bidirectional relationships between sexual subjectivity and sexual behavior. In addition, no previous study has examined the timing of pubertal development and sexual subjectivity. Evidence suggests that adolescents that mature earlier, compared to their peers, form romantic relationships earlier and experience sexual behaviors earlier [11,55]. In the current and past research, the link between age and sexual subjectivity has been relatively weak [34]. Substituting timing of pubertal status for age may provide more evidence regarding individual characteristics that influence differences in sexual subjectivity and its development over time.	2	1	Third, although longitudinal data can be a quite powerful tool for understanding change and development, the current analyses squander some of that power. The regression results presented in Table 2 test whether sexual behaviors that participants already experienced by Time 1 predicted facets of sexual subjectivity at Time 2, controlling for sexual subjectivity at Time 1. Why would behavior that has already happened contribute to a re-ordering of individuals over the course of the next year? It seems that a more interesting and direct test of the longitudinal effects of sexual behavior on sexual subjectivity would test whether new sexual behaviors (that is, sexual behaviors experienced between Time 1 and Time 2) predict change in sexual subjectivity from Time 1 to Time 2. Moreover, the reverse paths -- from sexual subjectivity to future behavior -- are also not tested in this paper. More generally, I think the putative timescale of effects of behavior on sexual subjectivity needs more theoretical elaboration. Are behaviors in early adolescence (before age 16), for example, expected to continue to influence the trajectory of sexual subjectivity regardless of later sexual behavior? That seems to be the model implied by the analyses, but it is never explicitly specified.	1	2	bs6010004_perova	1
+We have trimmed the paper to focus on two of the five elements of sexual subjectivity and anticipated that these two elements would show different results. Thus, we have maintained the separate analyses of these two elements. Please see the added Hypotheses on p. 5-6.	2	1	Fourth, each analysis is conducted for each subscale separately, but (with the exception of sexual body-esteem) the scales are consistently (if moderately) intercorrelated. I think it would be informative to test whether associations with age or sexual experience group are operating through a general underlying factor of sexual subjectivity versus are unique to specific facets. The shift to a latent factor approach would have the added benefit of allowing the authors to use FIML to account for missing data at T2, and they could thus use their entire sample.	1	2	bs6010004_perova	1
+We have removed these analyses from the paper in an attempt to reduce the number of analyses and streamline the entire paper.	2	1	Fifth, one set of results seems quite contradictory: sense of entitlement to sexual self-pleasure decreased, on average, from Time 1 to Time 2, but sense of entitlement to sexual self-pleasure was positively correlated with age. How do the authors make sense of these result, as participants are one year older at Time 2? This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.	1	2	bs6010004_perova	1
+Thank you for this suggestion. The reference has been added to the paper.	2	1	"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion."	1	2	bs6010004_perova	1
+"The “inherent affinity” is more appropriate, and the term is updated in the manuscript. Revision in Page 1 Line 28:“The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2].” Point 2:"	2	1	The definition of biophilia is described as an “inherent love” toward nature. While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.	1	2	buildings12040417_makarova	1
+In-text citation in Page 1 Line 44: “Some scholars summarized and classified the natural design features into biophilic design frameworks to guide design activities [25, 39, 40, 50, 51, 52].” Three references are added in the References List: Ulrich, 1993. Biophilia, biophobia, and natural landscapes. The Biophilia hypothesis. USA: Island Press: Washington, D.C. Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. 6, 2332-2343. Ko et al., 2021. A Window View Quality Assessment Framework. Leukos. 1-26. DOI: 10.1080/15502724.2021.1965889. Ko et al., 2021.	2	1	P1, L39-41: Please provide references to these frameworks. Reading further to page 3, I believe these are the 24 biophilic design attributes [ref. 25,39], and the 14 patterns of biophilic design [ref.40]. Further references around the biophilic concept could also be provided, e.g.: Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. Ulrich, 1993. Biophilia, biophobia, and natural landscapes. Ko et al., 2020. A window view quality assessment framework. LEUKOS. The latter reference reviewed many international standards that advocate nature and biophilic design for view and building spaces, with examples given to the Singapore context. This somewhat overlaps with my next comment.	1	2	buildings12040417_makarova	1
+There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays. And the issue raised by the authors is that these biophilic design frameworks (design guidelines or standards) are general design frameworks (design guidelines or standards) which can be applied to all building typologies (i.e., residential buildings, workplaces, retails, etc). Further research needs to be conducted to develop the design guideline specific for workplace. The sentence was rewritten to demonstrate this argument. Revision in Page 2 Line 56-61: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guidelines and assessment methods are necessary.” Point 4:	2	1	P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment. WELL v2 has several features for Nature and Mind, and Biophilia – Parts I and II, with quantitative assessment methods provided. Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief. If the authors agree with this, perhaps this could be revised here to reflect this. Other standards likely incorporate biophilic elements in building architecture, and could be worth highlighting. The general issue raised by the authors do not necessarily imply a lack of guidelines for biophilic design, since there are several readily available, but may point toward prioritisation or emphasis of criteria to meet certain varying expectations, which was alluded to on lines 52-53.	1	2	buildings12040417_makarova	1
+We agree with this comment. Apart from the design evaluation, POE is also one of the mainstream research methods that can effectively diagnosing operation problems. The description and citations were inserted in the updated manuscript. Revision in Page 2 Line 77 ~ 80: “Moreover, from the perspective of building operation, the POE results also provide evaluation and feedback from occupants to the stakeholders and building managers on workplace biophilic design. Since POE is one of the mainstream research methods that can effectively diagnosing operation problems [54, 55, 56].” Citations are added in the Reference list: Graham, L.T., Parkinson, T., Schiavon, S., 2021. Lessons Learned from 20 years of CBE’s Occupant Surveys. Buildings and Cities 2(1):166-184. DOI: 10.5334/bc.76 Kent, M., Parkinson, T., Kim, J., Schiavon, S., 2021. A Data-Driven Analysis of Occupant Workspace Dissatisfaction. Building and Environment 205, 108270 Cheung, T., Schiavon, S., Graham, L.T., Tham, K.W., 2021. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. Building and environment (188). DOI：10.1016/j.buildenv.2020.107443 Point 5:	2	1	P2, L63: Although I wouldn’t completely rule this out, POE surveys may not always provide feedback to the architect, since they are implemented post design-stage and the building would be operated by facility management or the owner. In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues. Lessons learned from 20 years of CBE’s occupant surveys. Building & Cities. Kent et al., 2021. A data-driven analysis of occupant workspace dissatisfaction. Building and Environment. Cheung et al. 2021. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. Recently POE studies, also using office data, advocate this as benefit to their implementation, albeit not necessarily being the only reason: Graham et al., 2020.	1	2	buildings12040417_makarova	1
+The sentence was rewritten in the updated version. Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].”	2	1	P3, L92: I think refers to “has helped” given the five decades predating this.	1	2	buildings12040417_makarova	1
+Yes, the existing psychological scales are well-developed, but scales for investigating “workplace biophilic design” had not been developed before. Hence, in this study, we develop a method that focus on evaluating the biophilic design elements in workplace. The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE). The questions in the first and the second subscales are referred to the validated scales--The World Health Organization Quality of Life (WHOQOL) and the nature relatedness (NR). The third scale is focus on evaluation on the biophilic design elements. Hence, the questions in the final section are designed based on the selection of the biophilic design elements/attributes that typically applied in the office design, which are not mentioned in the previous scales.	2	1	P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation. Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32. Therefore, it would be more appropriate to use psychological scales (e.g., PANAS or psychological restoration), instead of design orientated question or survey. If the authors agree with this, this aspect could be revised.	1	2	buildings12040417_makarova	1
+The method of selection of the biophilic design attributes/patterns for workplace is: ŸStep one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks. ŸStep two, we neglect the patterns which are not representative in office environment (please find the detailed explanations in the respond for Comment #8) and specify the selected design patterns to nine biophilic design attributes. The detailed process is shown in Section 2.1 and Figure 1. ŸStep three, verify the selection of the nine biophilic attributes matches the validated eight factors that affect workers’ satisfaction and productivity (please find the detailed explanations in the respond for Comment #11). In terms of the reviewer think that the listed attributes span across different domains and the communal features are not that apparent, we believe it is due to the research perspective of biophilic design is different from the perspectives of building science and traditional POE studies (please find the detailed explanations in the respond for Comment #9).	2	1	Something I felt would useful would at the beginning would be a clear definition for what “biophilic attributes” refers to. Figure 1 provides some insights into this, but these listed attributes span across different domains and the communal features are not that apparent. This becomes an issue later, since some aspects referring to biophilic design become unclear.	1	2	buildings12040417_makarova	1
+The authors gave the reason after the sentence in Page 4, Line 173-174 of the original manuscript “Second, the authors neglect seven design patterns from the 14 Patterns of Biophilic Design which are not representative of the workplace design.” to explain why the seven of the patterns from the 14 patterns of biophilic design were discarded: For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended but not demonstrative in this building typology (workplace). Those are usually applicable in other building typologies, such as hotels or residential. To further explain why these patterns are recommended but are discarded: 1) First, these design patterns (i.e., the seven discarded patterns of the 14 biophilic design patterns) are recommended because they are proofed that benefits health. 2) However, these patterns are not common in most offices. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”. 3) Therefore, we only included those biophilic design patterns that relatively easy to apply in the workplace (e.g., greenery, natural light, artworks), and discard those which are not representative in an office design. And the sentences are revised to further explain the discard of the seven patterns. Please see the revised contents in the updated manuscript below: Revision in Page 5 Line 191-197: “For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended because they are proofed that benefits health. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”.” Point 9:	2	1	P4, L173-174: The authors state that seven of the patterns from the 14 patterns of biophilic design were discarded. If this was the case, then please better articulate its overarching utility in this study, considering that half of the patterns were not relevant to the research scope.	1	2	buildings12040417_makarova	1
+We believe that there is no conflict between the different classifications. The same parameters (e.g., daylight, thermal comfort, and air-quality, office layout and building form) can be classified in different classifications (i.e., the traditional POE frameworks and the biophilic design frameworks) by different perspectives. 1) From perspective of building science, building performance, and traditional POE, these parameters (e.g., daylight, thermal comfort, and air-quality) are considered as indoor environment parameters, and office layout and building form are considered a physical and architectural parameters. 2) On the other hand, from the perspective of biophilia and biophilic design, (e.g., factors workers’ satisfaction and productivity), these parameters are re-classified and defined as the factors affecting workplace health. Both classifications validated by previous literatures.	2	1	P4, L180-183: In traditional POE studies and general building science research, daylight, thermal comfort, and air-quality, would be considered as indoor environment parameters (as examples, please see refs. in comment #4), while office layout and building form would be considered a physical and architectural parameters. Reading further to page 5, lines 189-192, the authors begin to suggest to this, but referred to them and others indoor environmental parameters as factors for the workplace. I would suggest better rationalising the connections between the nine design parameters to biophilia to make these more overt.	1	2	buildings12040417_makarova	1
+We revised the Figure 1. The connections between 24 Biophilic Design Attributes (Column A) and 14 Patterns of Biophilic Design (Column B); The nine biophilic design attributes for the workplace (Column C). First, to simplify the image, we put all the definitions into a new table in the appendix (Appendix A. Definitions of biophilic design attributes and patterns). In case that there are some readers are not familiar with the biophilic design attributes or patterns: Revision in Page 4 Line 164: “These two biophilic frameworks are chosen as research references (definitions of the at-tributes and patterns are in Appendix A).” Appendix A in Page 22: “Appendix A. Definitions of biophilic design attributes and patterns” Second, we fill the correlated boxes with same solid colours to make them more recognizable: Revision in Page 6 Line 207: Point 11:	2	1	Please consider simplifying the figure. Figure 1: The image presenting all the linkages is very interesting and is worth emphasising, but contains an overwhelming degree of information, and the text and line sizes are too small for readership. For example, some text boxes many not need further explanation (e.g. presence of water); also the lines connecting column A to the same patterns in column 4 could be colour coordinated.	1	2	buildings12040417_makarova	1
+Table 2 made a comparison between the selected nine biophilic design patterns for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace. These overlapped factors highlight the nine biophilic design attributes that are critical to the office design. Hence, the validation of the selection of the nine biophilic attributes are proofed by the previous literature. Sentences are added in the paragraph to clarify the demonstration. Revision in Page 6 Line 215-220: “Table 2 made a comparison between the selected nine biophilic design attributes for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. These overlapped factors highlight the nine biophilic design at-tributes that are critical to the office design. The validation of the selection of the nine biophilic attributes are proofed by the previous literature [37].” Point 12:	2	1	Table 2: Please consider providing further explanations for this table. It was not clear what the authors wanted to show.	1	2	buildings12040417_makarova	1
+1) The reasons why the two offices are selected for investigation: a) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices. Revision in Page 9 Line 277-280: “The two offices have similar features: 1) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices.” 2) The supplemental Information (i.e., temperature, number of employees) are added in the revised version. Revision in Page 9 Table 5: Table 5. The structure is re-constructed in the revised version. The original Section 3.1.	2	1	P8, L245: Please specify why these two offices were of interest (e.g., were they comparable or had specify architectural features worthy of study). If possible, please provide more characteristics (e.g., size, floor area, furniture layout (e.g., open-plan or enclosed), etc.) for each office. Later (P10, L299), it says 201 questionnaires were collected, with 161 occupants taking part in the Singaporean office. This led me to believe that this office was much larger than the building studied in China. An image showing the indoor conditions and outdoor façade for each might be beneficial. Many of the explanations found in section 3.1 could be moved into this part of the manuscript, since they many describe and show the existing office conditions and to do necessarily form part of the main results.	1	2	buildings12040417_makarova	1
+The means (SD) are only applied when the assumption of normality applies for the datasets. Revision in Page 18-20 Table 9: Table 9.	2	1	Section 3.3. Although I appreciated the thoroughness to which the descriptive statistical was explained, I wasn’t convinced the mean was the best indicator for the data, considering that evaluation scores were collected on a 5-point scale and not a continuous linear one. In-lieu of the mean, please consider using the median and inter-quartile range as the central tendency and dispersion indicators. Figures 1 and 4 can be removed, as the assumption of normality no longer applies (also on P15, L399-400), or replaced with boxplots.	1	2	buildings12040417_makarova	1
+The Cronbach’s Alpha with an Alpha>0.6 considered acceptable internal consistency in this study. The statement and the references are added in the revised version. Revision in Page 13 Line 381-383: “The Cronbach’s α coefficient value of the main scale is 0.72, while those of the sub-scales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).” Citations are added in the Reference list: Morgan, P. J., Cleave‐Hogg, D., DeSousa, S., Tarshis, J., 2004. High‐fidelity patient simulation: validation of performance checklists. British Journal of Anesthesia, Volume 92, (3) 388–392. Cronbach, L.J., 1951. Coefficient Alpha and the Internal Structure of tests, Psychometrika. Vol. 6. 3. 297-334.	2	1	Table 8: Please consider applying benchmarks for what constitute reasonable levels for internal consistency, when using the Cronbach’s Alpha (e.g., α>0.7): Please see, for example: Taber, 2018. The use of Cronbach’s Alpha when developing and reporting research instruments in science education. Research in Science Education. Tavakol et al. Making sense of Cronbach’s Alpha. International Journal of Medical Education.	1	2	buildings12040417_makarova	1
+1) Abbreviated labels referring to the actual questions are added in the Figure 3 (Figure 2 in the revision). But 2) we keep the original percentages (round to one decimal place). We think it is not hard for the readers to understand, and the original percentages (round to one decimal place) is more accurate. Revision in Page 17 Line 431:	2	1	Figure 3. The plot is well presented. A few minor notes for improvement: 1) Please consider adding short or abbreviated labels referring to the actual question, instead of codes (e.g., GH3-Q10). This would make it easier for the reader; 2) Round the percentages to the nearest whole number.	1	2	buildings12040417_makarova	1
+Thanks for the reminder, the error has been corrected in the revised version. Revision in Page 21 Line 473: “According to Table 10, Pearson correlations indicate that…” Point 17:	2	1	P17, L434: Please correct the unfortunate citation error on this line.	1	2	buildings12040417_makarova	1
+Spearman’s correlation test was conducted, and the revised contents are shown in the updated version. Revision in Page 17 Line 440: “Second, Spearman's correlation analysis is utilized to examine the correlation between three subscales.” Revision in Page 21 Line 472-483: 4.2. Intercorrelation between the three subscales (GH, NR, BDE) “According to Table 10, Spearman's correlations indicate that works' nature relatedness (NR) was positively correlated with self-evaluated GH (r = .264**, p < .01). This result also confirms the previously obtained results that people who had a higher evaluation in nature relatedness are also had a higher evaluation on their health. When the occupants feel that they have a strong sense of relationship with nature, it is observed that the biophilic environment would have positive impacts on their health. More importantly, significant correlation is also found in between biophilic design evaluation and self-reported health (GH), r=.270**, p < .01, indicating that office biophilic design has positive values on workers’ psychological health.” Table 10. Intercorrelations between responses of three subscales. Correlation is significant at the 0.01 level (2-tailed) Point 18: P18, L448-450: Please check whether the sentence is accurate and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3.	2	1	P18, section 4.2: Similar to comment #13, the data may be more suited to a Spearman’s correlation test, instead of the Pearson’s correlation coefficient. Due to the reasonable size of the dataset collected, it may not change the interpretation, but would help improve the analytical rigour.	1	2	buildings12040417_makarova	1
+The above contents have been corrected in the revised version. Point 19: While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavors, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work but did really emerge from the final section of their work in the same way it was emphasized in the abstract.	2	1	The sentence reads: Homogenous subsets with significant discrepancies (differences?) across subsets, leading to no significant differences across subsets. The above is not easy to grasp. If the information is accurate, please consider amending this to make this clearer. P18, L448-450: Please check whether the sentence is accurate, and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3.	1	2	buildings12040417_makarova	1
+The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 514-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Author Response File: Author Response.docx	2	1	While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavours, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work, but did really emerge from the final section of their work in the same way it was emphasised in the abstract.	1	2	buildings12040417_makarova	1
+In this revised version, we further explain the lack of scales for biophilic design evaluation. Please find the revised sentences as below: Revision in Page 3 Line 122-128: “Actually, these are standard research methods of investigative POE; nevertheless, there are no existing POE scales that focus on biophilic design in the workplace. According to the research objectives of this study (i.e., evaluate the subjective health impacts of biophilic design in workplace), we need to refer the well-developed scales from other disciplinary (e.g., Environmental Psychology). And finally, a scale that integrated health evaluation and building environment evaluation (i.e., POE) is developed for investigation.” Point 2: In consideration to their response #8, I understood and agreed with why half the seven of the 14 papers were discarded, but it did not provide much insight into why the Patterns of Biophilic Design was still used.	4	1	For their response to comment #6, while I generally agreed with the authors, I felt that it didn’t quite address what I tried to originally convey. POE scales do not likely target biophilic elements in architectural designs directly, since a direct question (e.g., how satisfied are you with the biophilic features) may not accurately depict every beneficial nuance they offer (e.g., psychological recovery). Mayer’s connectedness to nature scale) to help measure them. My generally feeling is that scales, from other domains (i.e., outside of POE studies) have been adopted for this reason, and a short sentence explaining the rationale supporting the lack of scales for biophilic design could be provided to briefly mentioned this. Instead studies may elect to use, for example, other relatable scales, which may not have originally been designed for POE surveys (e.g.	3	2	buildings12040417_makarova	1
+In the revised version, sentences are added to further explain that there are too few framework available, and these two frameworks are the mainstream biophilic design frameworks that are widely applied in the practical biophilic design projects. Revision in Page 3 Line 122-128: “The 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design are the two mainstream biophilic design which are widely applied in the practical biophilic design projects. Hence, these two frameworks are the most suitable to be selected as the basis of this experiment.”	4	1	If this was the case, perhaps this could be succinctly mentioned. In consideration to their response #8, I understood and agreed with why half the seven of the 14 papers were discarded, but it did not provide much insight into why the Patterns of Biophilic Design were still used. A more useful example, which may have elucidated this more, was whether are there too few frameworks available, and/or none others could have been used.	3	2	buildings12040417_makarova	1
+"1) Further explanations of “Biophilia” are added in the Introduction. Revision in Page 1 Line 27-29: “The term ""Biophilia"" is evolved from human evolution research and is coined to describe humans' inherent love affinity for the living things in the natural world [1,2]. It explained why we prefer nature because it is an instinct deeply rooted in the human brain.” 2) Literatures are added in the revised version to combine “workplace design” and “Biophilic Design”. Revision in Page 2 Line 48-53: “Workplace is one of the typologies that attracts the attentions of researchers. Scholars who research the relationship between the built environment and health found that the environment not merely directly or indirectly affects human health but also affects their work and study performance [59]. Studies proofed that biophilic design benefits workers’ health and productivity in an office environment [60, 61, 62, 63].” Citations are added in the Reference list: Derek, C. C., 2003. Environmental Quality and the Productive Workplace. In C. J. Anumba (Ed.), Innovative Developments in Architecture, Engineering and Construction. Rotterdam: Millpress Science Publishers. Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021."	2	1	"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_makarova	1
+"Observation Results-Biophilic Design Attributes in the Selected Offices is moved to be Section 2.3.2. It is because the 2.3.1 is the selection of office, and the Section 2.3.2 provides observation details of the selected offices. After re-construction, Section 3. Questionnaire Results focus on illustrating the questionnaire results. Hence, all the means (SD) are replaced as medians (IQR). And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version. Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8). According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min. 20 to max. 100). Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR. The median (IQR) value of the POE scale BDE is evaluated as 42.00 (5.00) (range of total value: min. 12 to max. 60). Table 8. Medians, Interquartile Range (IQR), Mean, standard deviation, and α coefficient values of workers’ evaluation based on HWBO, GH, NR, and BDE. Comparison of independent variables (Gender, Age, Educational Levels, Weekly Work Hours, Daily Sedentary Time, Work Desk Locations, Working Years, Office Locations) on self-reported GH, NR, and BDE. Thanks for your suggestion. The figures and pictures have been adjusted in the revised manuscript. The Section 3 Questionnaire Results and Section 4 are rewritten in the revised version: a) the statistical analysis is modified; b) to reduce elaboration in the text. Please kindly refer the following revisions: 1) The original Section 3.1. 2) In the revised version, the formulars in Section 3.3 2. Quantitative Results of Impacts of Biophilic Design for Workplace are removed due to the statistical analysis is modified: The means (SD) are only applied when the assumption of normality applies for the datasets. Revision in Page 18 Table 9: Table 9. *p<.05, The significance level is .05.3) Elaboration for the Figure 2. Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement) is reduced, because the illustrations are obvious in the figure. Revision in Page 15-17 Line 399-433: The analysis of individual items provides more details into the works' responses. As can be seen in percentage responses for individual items (Figure 2 in revision), most of the responses are distributed in the items ""Neutral"" and ""Agree"". The questionnaire results reveal that the employees from the understudy companies hold a relatively positive opinion on wellbeing, nature-relatedness, indoor environmental quality, and biophilic design for their health promotion. According to the arrangement, at the top of this, stacked graph are the evaluation of satisfaction of the work capacities and relationships in the workplace. About 73.2% of respondents agree that the workers of the companies under investigation are satisfied with their work capacity (GH3-Q10) and relationships (GH4-Q11). In the subscale nature relatedness, 62.2% and 61.7% of workers responded (agree/strongly agree) that their actions affect the environment (NR2-Q13), and they take notice of the wildlife in their daily lives (NR3-Q14). Nevertheless, only 47.8% of them selected agreed/strongly agree regarding the statements that their ideal spot for vacation would be a wilderness area (NR1-Q12). Regarding the POE results in the subscale BDE, 63.2% of workers agree/strongly agree that the natural light is an essential biophilic attribute and their offices are bright (BDE2-Q17). Furthermore, 60.7 % of the workers agreed that introducing natural colors in the office benefits workplace health and wellbeing (BDE11-Q26). More than 60 percent (approximately 60.7%) of respondents believe that greenery is a biophilic design that benefits office wellbeing (BDE6-Q21). Their feedback would be valuable for designers to note that application of the biophilic design attributes in the office design can enhance the experiences and evaluations of workers. The quantitative results of the questionnaire survey demonstrate that the workers agree that the biophilic design attributes in the office have positive effects on their subjective wellbeing. Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement)."	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_makarova	1
+Thank you for your comments. These considerations are the limitations of this study. 1) these two cases are limited in representing all the workplace biophilic designs. As mentioned in the Conclusion, in the future study, we will include more offices and locations as experiment samples. In the future study, the research scope should be narrow down for intensive investigation.	2	1	The result of biophilic design frameworks for specific workplace typology is of certain value. The location of the workplace selected in this article is limited, the related biophilic factors are scientifically screened, and the researchers have made a detailed classification study. However, the huge research scope has certain obstacles to the relevant results, and the general and targeted conclusions will be worse.	1	2	buildings12040417_makarova	1
+Thank you for your suggestion. The description of the necessity of research is highlight in the revised version. Please kindly refer the following revisions: Revision in Page 2 Line 56-66: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guide-lines and assessment methods are necessary. Additionally, the effectiveness of such design in practical design projects for user wellbeing still requires confirmation. More importantly, building typology-based biophilic design guidelines should be appropriately developed because it would affect the designer's prioritization of design attributes selection in design practice.” Point 6:	2	1	It is best to supplement and describe the necessity of research. At the same time, it will be more complete if the results are reflected in the summary.	1	2	buildings12040417_makarova	1
+The different genders and different ages are included in the study. The age ranges included 21-25, 26-35, 36-45, 46-60. The detailed description of the demographic information is in Section 3.1. Demographic Information.	2	1	The object of the questionnaire should be composed of people with different ages and genders, which is best explained in the article.	1	2	buildings12040417_makarova	1
+b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices. The weighting results of this study would be especially applicable to the workplace typology. The 14 Patterns of Biophilic Design has a broader range of usage for all building typologies and is more suitable for general design applications. Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design. These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results). c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration. d) The correlation results support the importance of biophilic design from the user perspectives. There is a significant correlation between office biophilic design and self-reported health of employees (r=.270**, p < .01). 2) The investigative POE studies evaluated the self-reported health (GH), nature-relatedness (NR), and biophilic design in the workplace (BDE). The objective of the study is to evaluate the typical biophilic design attributes in office environment and the correlation between biophilic design and office health. Hence, the research scope is relatively extensive. 1) The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” 2) The Conclusion is reconstructed and a separate section-- Section 6. Limitation and Future Studies is added in the revised version: Revision in Page 23 Line 542-551: “6. Limitation and Future Studies There are limitations in this study, first, these two cases are limited in representing all the workplace biophilic designs due to the sample size limitations. Further studies could include more offices and locations as experiment samples. In the future study, the research scope should be narrow down for intensive investigation.” Point 8:	2	1	Adding some explanations, examples and data comparison to the conclusion will be more intuitive and convincing, just as discussed earlier in the article. Perhaps this makes this article more complete and credible. At the same time, the conclusion only summarizes the article, and lasks discussions and explanations for the future research direction.	1	2	buildings12040417_makarova	1
+The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology. And the questionnaire can be applied in future biophilic design research for investigation. The applications of the results are highlighted in Section 1.4. Objectives and in Section 5 Conclusion: In Section 1.4. Objectives, the three practical applications of the results are mentioned: first, a POE questionnaire for assessing the biophilic design for workplace accounting for health and wellbeing. Second, the study will provide a new biophilic design guidelines for workplaces, which can effectively assist researchers and designers to improve office biophilic design practices and decision-making on design attributes selection. The Conclusion is rewritten to highlight the contribution and implementation of the results: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. … “c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration.” … “e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Point 9: It is better to supplement the temperature, humidity, and other parameters of the selected office in the part of the experiment, so as to facilitate readers' reference rather than just giving the location.	2	1	The theoretical part of the presentation is extensive, but the application part is relatively poorly described.	1	2	buildings12040417_makarova	1
+Basic information of the understudy offices. The supplemental Information (i.e., office temperature, number of employees) are added in the revised version. Revision in Table 5: Table 5.	2	1	It is better to supplement the temperature, humidity and other parameters of the selected office in the part of the experiment, so as to facilitate readers' reference rather than just giving the location.	1	2	buildings12040417_makarova	1
+We agree with the comment. A quantitative study for indoor workplace biophilic design to improve health and productivity performance. Journal of Cleaner Production. 324, 129168. Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018. Physiological and cognitive performance of exposure to biophilic indoor environment. Build. Environ. 132, 255–262. Yin, J., Arfaei, N., MacNaughton, P., Catalano, P.J., Allen, J.G., Spengler, J.D., 2019. Effects of biophilic interventions in office on stress reaction and cognitive function: a randomized crossover study in virtual reality. Indoor Air 29, 1028–1039. https:// doi.org/10.1111/ina.12593. Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020. Effects of biophilic indoor environment on stress and anxiety recovery: a between-subjects experiment in virtual reality. Environ. Int. 136, 105427. https://doi.org/10.1016/j. envint.2019.105427. Previous studies proofed that biophilic design in workplaces benefits both psychological and physiological health [60, 61, 62, 63]. This study is a questionnaire survey that focus on investigating the subjective evaluation on workplace biophilic design and of workers. Physiological measurements will be included in the future study to investigate the impacts of the psychological factors on physiological comfort. References: Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021.	2	1	The author can think about the impact of such a pro biological design model on the psychology of different experimental personnel. I think psychological factors will also affect human physiological comfort.	1	2	buildings12040417_makarova	1
+Thank you for your comment. Figure 1 is updated in this version.	4	1	Figure 1 should be improved regarding the arrow.	3	2	buildings12040417_makarova	1
+All the photos in Table 6 are taken by the designers. And we added an annotation under Table 6.	4	1	Photo in table 6 should check the copyright of these photo.	3	2	buildings12040417_makarova	1
+"Thank you for your suggestions. The references are added in the manuscript. In-text citations in Page 1 Line 27-28: “The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2,64].” In-text citations in Page 1 Line 40: “…the modern built environment' as 'Biophilic Design' [24,25,65].” In-text citations in Page 2 Line 69-70: “…occupant satisfaction, health, and wellbeing after occupancy of buildings [27, 28, 67].” Citations are added in the Reference list: Parsaee, M., Demers, M. H. C., Potvin, A., Hébert, M., Lalonde, J.F., Window View Access in Architecture: Spatial Visualization and Probability Evaluations Based on Human Vision Fields and Biophilia. 2021. Buildings, 11(12), 627. https://doi.org/10.3390/buildings11120627 Mollazadeh, M., Zhu, YM., Application of Virtual Environments for Biophilic Design: A Critical Review. 2021. Buildings, 11(4), 148; https://doi.org/10.3390/buildings11040148 Gillis, K., Gatersleben, B., 2015. A Review of Psychological Literature on the Health and Wellbeing Benefits of Biophilic Design. Buildings. 5(3), 948-963. https://doi.org/10.3390/buildings5030948 Candido, C., Chakraborty, P., Tjondronegoro, D., 2019. The Rise of Office Design in High-Performance, Open-Plan Environments. 9(4), 100. https://doi.org/10.3390/buildings9040100 Author Response File: Author Response.docx"	4	1	"Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."	3	2	buildings12040417_makarova	1
+"The “inherent affinity” is more appropriate, and the term is updated in the manuscript. Revision in Page 1 Line 28:“The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2].” Point 2:"	2	1	The definition of biophilia is described as an “inherent love” toward nature. While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.	1	2	buildings12040417_perova	1
+Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. Ulrich, 1993. Biophilia, biophobia, and natural landscapes. Ko et al., 2020. A window view quality assessment framework. LEUKOS. In-text citation in Page 1 Line 44: “Some scholars summarized and classified the natural design features into biophilic design frameworks to guide design activities [25, 39, 40, 50, 51, 52].” Three references are added in the References List: Ulrich, 1993. Biophilia, biophobia, and natural landscapes. The Biophilia hypothesis. USA: Island Press: Washington, D.C. Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. 6, 2332-2343. Ko et al., 2021. A Window View Quality Assessment Framework. Leukos. 1-26. DOI: 10.1080/15502724.2021.1965889. Ko et al., 2021.	2	1	P1, L39-41: Please provide references to these frameworks. Reading further to page 3, I believe these are the 24 biophilic design attributes [ref. 25,39], and the 14 patterns of biophilic design [ref.40]. Further references around the biophilic concept could also be provided, e.g.: Bjørn et al., 2009. The latter reference reviewed many international standards that advocate nature and biophilic design for view and building spaces, with examples given to the Singapore context. This somewhat overlaps with my next comment.	1	2	buildings12040417_perova	1
+There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays. And the issue raised by the authors is that these biophilic design frameworks (design guidelines or standards) are general design frameworks (design guidelines or standards) which can be applied to all building typologies (i.e., residential buildings, workplaces, retails, etc). Further research needs to be conducted to develop the design guideline specific for workplace. The sentence was rewritten to demonstrate this argument. Revision in Page 2 Line 56-61: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guidelines and assessment methods are necessary.” Point 4:	2	1	P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment. WELL v2 has several features for Nature and Mind, and Biophilia – Parts I and II, with quantitative assessment methods provided. Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief. Other standards likely incorporate biophilic elements in building architecture and could be worth highlighting. The general issue raised by the authors do not necessarily imply a lack of guidelines for biophilic design, since there are several readily available, but may point toward prioritization or emphasis of criteria to meet certain varying expectations, which was alluded to on lines 52-53. If the authors agree with this, perhaps this could be revised here to reflect this.	1	2	buildings12040417_perova	1
+Lessons learned from 20 years of CBE’s occupant surveys. A data-driven analysis of occupant workspace dissatisfaction. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. We agree with this comment. Apart from the design evaluation, POE is also one of the mainstream research methods that can effectively diagnosing operation problems. The description and citations were inserted in the updated manuscript. Revision in Page 2 Line 77 ~ 80: “Moreover, from the perspective of building operation, the POE results also provide evaluation and feedback from occupants to the stakeholders and building managers on workplace biophilic design. Since POE is one of the mainstream research methods that can effectively diagnosing operation problems [54, 55, 56].” Citations are added in the Reference list: Graham, L.T., Parkinson, T., Schiavon, S., 2021. Buildings and Cities 2(1):166-184. DOI: 10.5334/bc.76 Kent, M., Parkinson, T., Kim, J., Schiavon, S., 2021. Building and Environment 205, 108270 Cheung, T., Schiavon, S., Graham, L.T., Tham, K.W., 2021. Building and environment (188). DOI：10.1016/j.buildenv.2020.107443 Point 5:	2	1	P2, L63: Although I wouldn’t completely rule this out, POE surveys may not always provide feedback to the architect, since they are implemented post design-stage and the building would be operated by facility management or the owner. In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues. Recently POE studies, also using office data, advocate this as benefit to their implementation, albeit not necessarily being the only reason: žGraham et al., 2020. Building & Cities. Kent et al., 2021. Building and Environment. Cheung et al. 2021.	1	2	buildings12040417_perova	1
+The sentence was rewritten in the updated version. Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].” Point 6:	2	1	P3, L92: I think refers to “has helped” given the five decades predating this.	1	2	buildings12040417_perova	1
+Yes, the existing psychological scales are well-developed, but scales for investigating “workplace biophilic design” had not been developed before. Hence, in this study, we develop a method that focus on evaluating the biophilic design elements in workplace. The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE). The questions in the first and the second subscales are referred to the validated scales--The World Health Organization Quality of Life (WHOQOL) and the nature relatedness (NR). The third scale is focus on evaluation on the biophilic design elements. Hence, the questions in the final section are designed based on the selection of the biophilic design elements/attributes that typically applied in the office design, which are not mentioned in the previous scales.	2	1	P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation. Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32. Therefore, it would be more appropriate to use psychological scales (e.g., PANAS or psychological restoration), instead of design orientated question or survey. If the authors agree with this, this aspect could be revised.	1	2	buildings12040417_perova	1
+The method of selection of the biophilic design attributes/patterns for workplace is: ŸStep one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks. ŸStep two, we neglect the patterns which are not representative in office environment (please find the detailed explanations in the respond for Comment #8) and specify the selected design patterns to nine biophilic design attributes. The detailed process is shown in Section 2.1 and Figure 1. ŸStep three, verify the selection of the nine biophilic attributes matches the validated eight factors that affect workers’ satisfaction and productivity (please find the detailed explanations in the respond for Comment #11). In terms of the reviewer think that the listed attributes span across different domains and the communal features are not that apparent, we believe it is due to the research perspective of biophilic design is different from the perspectives of building science and traditional POE studies (please find the detailed explanations in the respond for Comment #9).	2	1	Something I felt would useful would at the beginning would be a clear definition for what “biophilic attributes” refers to. Figure 1 provides some insights into this, but these listed attributes span across different domains and the communal features are not that apparent. This becomes an issue later, since some aspects referring to biophilic design become unclear.	1	2	buildings12040417_perova	1
+The authors gave the reason after the sentence in Page 4, Line 173-174 of the original manuscript “Second, the authors neglect seven design patterns from the 14 Patterns of Biophilic Design which are not representative of the workplace design.” to explain why the seven of the patterns from the 14 patterns of biophilic design were discarded: For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended but not demonstrative in this building typology (workplace). Those are usually applicable in other building typologies, such as hotels or residential. To further explain why these patterns are recommended but are discarded: 1) First, these design patterns (i.e., the seven discarded patterns of the 14 biophilic design patterns) are recommended because they are proofed that benefits health. 2) However, these patterns are not common in most offices. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”. 3) Therefore, we only included those biophilic design patterns that relatively easy to apply in the workplace (e.g., greenery, natural light, artworks), and discard those which are not representative in an office design. And the sentences are revised to further explain the discard of the seven patterns. Please see the revised contents in the updated manuscript below: Revision in Page 5 Line 191-197: “For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended because they are proofed that benefits health. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”.” Point 9:	2	1	P4, L173-174: The authors state that seven of the patterns from the 14 patterns of biophilic design were discarded. If this was the case, then please better articulate its overarching utility in this study, considering that half of the patterns were not relevant to the research scope.	1	2	buildings12040417_perova	1
+We believe that there is no conflict between the different classifications. The same parameters (e.g., daylight, thermal comfort, and air-quality, office layout and building form) can be classified in different classifications (i.e., the traditional POE frameworks and the biophilic design frameworks) by different perspectives. 1) From perspective of building science, building performance, and traditional POE, these parameters (e.g., daylight, thermal comfort, and air-quality) are considered as indoor environment parameters, and office layout and building form are considered a physical and architectural parameters. 2) On the other hand, from the perspective of biophilia and biophilic design, (e.g., factors workers’ satisfaction and productivity), these parameters are re-classified and defined as the factors affecting workplace health. Both classifications validated by previous literatures.	2	1	P4, L180-183: In traditional POE studies and general building science research, daylight, thermal comfort, and air-quality, would be considered as indoor environment parameters (as examples, please see refs. in comment #4), while office layout and building form would be considered a physical and architectural parameters. Reading further to page 5, lines 189-192, the authors begin to suggest to this, but referred to them and others indoor environmental parameters as factors for the workplace. I would suggest better rationalizing the connections between the nine design parameters to biophilia to make these more overt.	1	2	buildings12040417_perova	1
+We revised the Figure 1. The connections between 24 Biophilic Design Attributes (Column A) and 14 Patterns of Biophilic Design (Column B); The nine biophilic design attributes for the workplace (Column C). First, to simplify the image, we put all the definitions into a new table in the appendix (Appendix A. Definitions of biophilic design attributes and patterns). In case that there are some readers are not familiar with the biophilic design attributes or patterns: Revision in Page 4 Line 164: “These two biophilic frameworks are chosen as research references (definitions of the at-tributes and patterns are in Appendix A).” Appendix A in Page 22: “Appendix A. Definitions of biophilic design attributes and patterns” Second, we fill the correlated boxes with same solid colours to make them more recognizable: Revision in Page 6 Line 207: Point 11:	2	1	Figure 1: The image presenting all the linkages is very interesting and is worth emphasizing, but contains an overwhelming degree of information, and the text and line sizes are too small for readership. Please consider simplifying the figure. For example, some text boxes many do not need further explanation (e.g., presence of water); also, the lines connecting column A to the same patterns in column 4 could be color coordinated.	1	2	buildings12040417_perova	1
+Table 2 made a comparison between the selected nine biophilic design patterns for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace. These overlapped factors highlight the nine biophilic design attributes that are critical to the office design. Hence, the validation of the selection of the nine biophilic attributes are proofed by the previous literature. Sentences are added in the paragraph to clarify the demonstration. Revision in Page 6 Line 215-220: “Table 2 made a comparison between the selected nine biophilic design attributes for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. These overlapped factors highlight the nine biophilic design at-tributes that are critical to the office design. The validation of the selection of the nine biophilic attributes are proofed by the previous literature [37].” Point 12:	2	1	Table 2: Please consider providing further explanations for this table. It was not clear what the authors wanted to show.	1	2	buildings12040417_perova	1
+1) The reasons why the two offices are selected for investigation: a) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices. Revision in Page 9 Line 277-280: “The two offices have similar features: 1) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices.” 2) The supplemental Information (i.e., temperature, number of employees) are added in the revised version. Revision in Page 9 Table 5: Table 5. Basic information of the understudy offices. The structure is re-constructed in the revised version. The original Section 3.1.	2	1	P8, L245: Please specify why these two offices were of interest (e.g., were they comparable or had specify architectural features worthy of study). If possible, please provide more characteristics (e.g., size, floor area, furniture layout (e.g., open-plan or enclosed), etc.) for each office. Later (P10, L299), it says 201 questionnaires were collected, with 161 occupants taking part in the Singaporean office. This led me to believe that this office was much larger than the building studied in China. An image showing the indoor conditions and outdoor façade for each might be beneficial. Many of the explanations found in section 3.1 could be moved into this part of the manuscript, since they many describe and show the existing office conditions and to do necessarily form part of the main results.	1	2	buildings12040417_perova	1
+We agree with the comment. The means (SD) are only applied when the assumption of normality applies for the datasets. According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min. 20 to max. 100). Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR. The median (IQR) value of the POE scale BDE is evaluated as 42.00 (5.00) (range of total value: min. 12 to max. 60). Table 8. Medians, Interquartile Range (IQR), Mean, standard deviation, and α coefficient values of workers’ evaluation based on HWBO, GH, NR, and BDE. Revision in Page 18-20 Table 9: Table 9. Comparison of independent variables (Gender, Age, Educational Levels, Weekly Work Hours, Daily Sedentary Time, Work Desk Locations, Working Years, Office Locations) on self-reported GH, NR, and BDE.	2	1	Section 3.3. Although I appreciated the thoroughness to which the descriptive statistical was explained, I wasn’t convinced the mean was the best indicator for the data, considering that evaluation scores were collected on a 5-point scale and not a continuous linear one. In-lieu of the mean, please consider using the median and inter-quartile range as the central tendency and dispersion indicators. Figures 1 and 4 can be removed, as the assumption of normality no longer applies (also on P15, L399-400), or replaced with boxplots.	1	2	buildings12040417_perova	1
+Thank you for your suggestion. The Cronbach’s Alpha with an Alpha>0.6 considered acceptable internal consistency in this study. The statement and the references are added in the revised version. Revision in Page 13 Line 381-383: “The Cronbach’s α coefficient value of the main scale is 0.72, while those of the sub-scales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).” Citations are added in the Reference list: Morgan, P. J., Cleave‐Hogg, D., DeSousa, S., Tarshis, J., 2004. High‐fidelity patient simulation: validation of performance checklists. British Journal of Anesthesia, Volume 92, (3) 388–392. Cronbach, L.J., 1951. Coefficient Alpha and the Internal Structure of tests, Psychometrika. Vol. 297-334.	2	1	Table 8: Please consider applying benchmarks for what constitute reasonable levels for internal consistency, when using the Cronbach’s Alpha (e.g., α>0.7): Please see, for example: Taber, 2018. The use of Cronbach’s Alpha when developing and reporting research instruments in science education. Research in Science Education. Tavakol et al. Making sense of Cronbach’s Alpha. International Journal of Medical Education.	1	2	buildings12040417_perova	1
+1) Abbreviated labels referring to the actual questions are added in the Figure 3 (Figure 2 in the revision). But 2) we keep the original percentages (round to one decimal place). We think it is not hard for the readers to understand, and the original percentages (round to one decimal place) is more accurate. Revision in Page 17 Line 431: Point 16:	2	1	Figure 3. The plot is well presented. A few minor notes for improvement: 1) Please consider adding short or abbreviated labels referring to the actual question, instead of codes (e.g., GH3-Q10). This would make it easier for the reader; 2) Round the percentages to the nearest whole number.	1	2	buildings12040417_perova	1
+Thanks for the reminder, the error has been corrected in the revised version. Revision in Page 21 Line 473: “According to Table 10, Pearson correlations indicate that…” Point 17:	2	1	P17, L434: Please correct the unfortunate citation error on this line.	1	2	buildings12040417_perova	1
+Spearman’s correlation test was conducted, and the revised contents are shown in the updated version. Revision in Page 17 Line 440: “Second, Spearman's correlation analysis is utilized to examine the correlation between three subscales.” Revision in Page 21 Line 472-483: 4.2. Intercorrelation between the three subscales (GH, NR, BDE) “According to Table 10, Spearman's correlations indicate that works' nature relatedness (NR) was positively correlated with self-evaluated GH (r = .264**, p < .01). This result also confirms the previously obtained results that people who had a higher evaluation in nature relatedness are also had a higher evaluation on their health. When the occupants feel that they have a strong sense of relationship with nature, it is observed that the biophilic environment would have positive impacts on their health. More importantly, significant correlation is also found in between biophilic design evaluation and self-reported health (GH), r=.270**, p < .01, indicating that office biophilic design has positive values on workers’ psychological health.” Table 10. Intercorrelations between responses of three subscales. Correlation is significant at the 0.01 level (2-tailed) Point 18:	2	1	P18, section 4.2: Similar to comment #13, the data may be more suited to a Spearman’s correlation test, instead of the Pearson’s correlation coefficient. Due to the reasonable size of the dataset collected, it may not change the interpretation, but would help improve the analytical rigor.	1	2	buildings12040417_perova	1
+Thank you for your comment. The above contents have been corrected in the revised version.	2	1	P18, L448-450: Please check whether the sentence is accurate and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3. The sentence reads: Homogenous subsets with significant discrepancies (differences?) across subsets, leading to no significant differences across subsets. The above is not easy to grasp. If the information is accurate, please consider amending this to make this clearer.	1	2	buildings12040417_perova	1
+The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 514-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology. And the questionnaire can be applied in future biophilic design research for investigation. b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices. The weighting results of this study would be especially applicable to the workplace typology. The 14 Patterns of Biophilic Design has a broader range of usage for all building typologies and is more suitable for general design applications. Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design. These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results). c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration. d) The correlation results support the importance of biophilic design from the user perspectives. There is a significant correlation between office biophilic design and self-reported health of employees (r=.270**, p < .01). e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Author Response File: Author Response.docx	2	1	While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavors, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work but did really emerge from the final section of their work in the same way it was emphasized in the abstract.	1	2	buildings12040417_perova	1
+"1) Further explanations of “Biophilia” are added in the Introduction. Revision in Page 1 Line 27-29: “The term ""Biophilia"" is evolved from human evolution research and is coined to describe humans' inherent love affinity for the living things in the natural world [1,2]. It explained why we prefer nature because it is an instinct deeply rooted in the human brain.”"	2	1	"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	1
+2) Literatures are added in the revised version to combine “workplace design” and “Biophilic Design”. Revision in Page 2 Line 48-53: “Workplace is one of the typologies that attracts the attentions of researchers. Scholars who research the relationship between the built environment and health found that the environment not merely directly or indirectly affects human health but also affects their work and study performance [59]. Studies proofed that biophilic design benefits workers’ health and productivity in an office environment [60, 61, 62, 63].” Citations are added in the Reference list:	2	1	"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	1
+Derek, C. C., 2003. Environmental Quality and the Productive Workplace. In C. J. Anumba (Ed.), Innovative Developments in Architecture, Engineering and Construction. Rotterdam: Millpress Science Publishers. Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021. A quantitative study for indoor workplace biophilic design to improve health and productivity performance. Journal of Cleaner Production. 324, 129168. Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018. Physiological and cognitive performance of exposure to biophilic indoor environment. Build. Environ. 132, 255–262. Yin, J., Arfaei, N., MacNaughton, P., Catalano, P.J., Allen, J.G., Spengler, J.D., 2019. Effects of biophilic interventions in office on stress reaction and cognitive function: a randomized crossover study in virtual reality. Indoor Air 29, 1028–1039. https:// doi.org/10.1111/ina.12593. Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020. Effects of biophilic indoor environment on stress and anxiety recovery: a between-subjects experiment in virtual reality. Int. 136, 105427. https://doi.org/10.1016/j. envint.2019.105427. 61.7% of workers responded (agree/strongly agree) that their actions affect the environment (NR2-Q13), and they take notice of the wildlife in their daily lives (NR3-Q14). 63.2% of workers agree/strongly agree that the natural light is an essential biophilic attribute and their offices are bright (BDE2-Q17). 60.7 % of the workers agreed that introducing natural colors in the office benefits workplace health and wellbeing (BDE11-Q26). Citations are added in the Reference list: 59.	2	1	"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	1
+Observation Results-Biophilic Design Attributes in the Selected Offices is moved to be Section 2.3.2. It is because the 2.3.1 is the selection of office, and the Section 2.3.2 provides observation details of the selected offices. After re-construction, Section 3. Questionnaire Results focus on illustrating the questionnaire results. Hence, all the means (SD) are replaced as medians (IQR). And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version. Thanks for your suggestion. The figures and pictures have been adjusted in the revised manuscript. The Section 3 Questionnaire Results and Section 4 are rewritten in the revised version: a) the statistical analysis is modified; b) to reduce elaboration in the text. Please kindly refer the following revisions: 1) The original Section 3.1. 2) In the revised version, the formulars in Section 3.3 2. Quantitative Results of Impacts of Biophilic Design for Workplace are removed due to the statistical analysis is modified: The means (SD) are only applied when the assumption of normality applies for the datasets.	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	1
+Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	1
+Revision in Page 18 Table 9: Table 9.	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	1
+Revision in Page 15-17 Line 399-433: The analysis of individual items provides more details into the works' responses.	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	1
+Figure 1 is updated in this version.	4	1	Figure 1 should be improved regarding the arrow.	3	2	buildings12040417_perova	1
+All the photos in Table 6 are taken by the designers. And we added an annotation under Table 6.	4	1	Photo in table 6 should check the copyright of these photo.	3	2	buildings12040417_perova	1
+Thank you for your suggestions. The references are added in the manuscript.	4	1	"Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."	3	2	buildings12040417_perova	1
+Subtotal resection is usually defined as a resection of 95-90%, with <90% defined as partial. We addended the submission in line 93-95 as well as table 4 as recommended.	2	1	How is subtotal resection defined in chordoma (% of volume remaining?)	1	2	cancers14040966_makarova	1
+We are grateful to the reviewer for catching these mistakes. Both volume reduction and pathology associated numbers are formatting errors and we addended the table to correct these mistakes.	2	1	What means Pathology 0 in table 4?	1	2	cancers14040966_makarova	1
+We agree with the reviewer that useful tools and techniques are available in different modalities and certainly surgeons dealing with various tumors utilize these as needed. We are fortunate to have access to this multi-modality image guided operating suite, and here we report on its use in specific rare tumors, chordomas and chondrosarcomas. We agree that the use of the PET-CT has no use in these cases, and we mentioned it only as part of the general introduction of the AMIGO suite. We addended the manuscript (line 260-264) as recommended	2	1	Thus it would be helpful for the authors to clarify this. The AMIGO suite has great capabilities, but when it comes to intraoperative imaging, it is CT and MRI scans that are made, and then using Brain Lab and possibly other software, that information is processed and redisplayed including 3D reconstructions.	1	2	cancers14040966_makarova	1
+We agree with the reviewer that it is the acquisition of multimodality (CT and MRI), with registration of preoperative imaging then intraoperative co-registration. We shared these steps in the “Operative and perioperative management” section (line 60).	2	1	This does not detract from the value of the technique, but it is important to clarify that what really is happening is a variety of preoperative imaging is imported and co-registered into the Brain Lab, and then intraoperative MR and occasionally CT data is again obtained and processed and analyzed.	1	2	cancers14040966_makarova	1
+As acknowledged in the limitation section and noted by the reviewer, the rarity of these tumors and the small number of patients in most series, including ours, limits analysis. Hence, we did not aim to generalize our outcome. Our focus was on how the use of these modern tools can facilitate maximum safe tumor removal which is considered a pivotal factor in the management of chordoma and chondrosarcoma.	2	1	It is therefore difficult to state that the results were clearly improved or to even compare to other series, without larger numbers and direct comparison or classification of individual tumor locations and characteristics in differing series.	1	2	cancers14040966_makarova	1
+Subtotal resection is usually defined as a resection of 95-90%, with <90% defined as partial. We addended the submission in line 93-95 as well as table 4 as recommended.	2	1	How is subtotal resection defined in chordoma (% of volume remaining?)	1	2	cancers14040966_perova	1
+We are grateful to the reviewer for catching these mistakes. Both volume reduction and pathology associated numbers are formatting errors and we addended the table to correct these mistakes.	2	1	What means Pathology 0 in table 4?	1	2	cancers14040966_perova	1
+We agree with the reviewer that it is the acquisition of multimodality (CT and MRI), with registration of preoperative imaging then intraoperative co-registration. We shared these steps in the “Operative and perioperative management” section (line 60).	2	1	when one reads it initially it would appear that there are many different modalities available for intraoperative imaging. However, the PET scan does not appear to have actually been used for intraoperative repeat imaging to assess resection, and fluoroscopy and 3D reconstructions using Brain Lab software are standard fare in many operating rooms. Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions. The authors appear to really only use CT and MRI intraoperatively, with the other modalities imported preoperatively into the Brain Lab navigational system. 3D reconstructions of newly acquired data is a capability of the software, and is available in many places. Thus it would be helpful for the authors to clarify this. The AMIGO suite has great capabilities, but when it comes to intraoperative imaging, it is CT and MRI scans that are made, and then using Brain Lab and possibly other software, that information is processed and redisplayed including 3D reconstructions. Making this clear is important for others who either use the technique or who want to do so. All that is needed is easy intra-operative access to CT and MR imaging and the ability to import and reconstruct the data real time. That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or). This does not detract from the value of the technique, but it is important to clarify that what really is happening is a variety of preoperative imaging is imported and co-registered into the Brain Lab, and then intraoperative MR and occasionally CT data is again obtained and processed and analyzed.	1	2	cancers14040966_perova	1
+As acknowledged in the limitation section and noted by the reviewer, the rarity of these tumors and the small number of patients in most series, including ours, limits analysis. Hence, we did not aim to generalize our outcome. Our focus was on how the use of these modern tools can facilitate maximum safe tumor removal which is considered a pivotal factor in the management of chordoma and chondrosarcoma.	2	1	The results, while admirable, cannot easily be compared to other series, as the numbers are small and each case has its own complex series of characteristics that make it unique. It is therefore difficult to state that the results were clearly improved or to even compare to other series, without larger numbers and direct comparison or classification of individual tumor locations and characteristics in differing series.	1	2	cancers14040966_perova	1
+As suggested, we analyzed the distribution of IRS (immunoreactivity score) across the cohort. In Figure 1 it can be clearly seen that there are two groups forming here, those with 0-4 and those with 6-12 IRS. In addition, we calculated the survival probability depending on the IRS (Figure 2). Patients with an IRS of 6 or higher have a similar bad prognosis compared to patients with an IRS below 5. A better survival correlates with lower MACC1 IRS. Patients with an IRS of 0 have the best prognosis. Based on these two analyses, the decision for cut-off value of 5 was made. Additionally, ROC curve analysis showed, that the IRS value of 5 has the highest Youden Index and is therefore the most suitable cut-off value. 0.408) (Figure 3 and Table 1). Looking at MACC1 expression in cell lines and tissue samples, the variability looks similar. Nevertheless, it must be considered that the investigation of the homogeneous cell lines is a purely quantitative analysis of the expression by Western plot and RT-PCR. The analysis of the tissue samples by immunoreactivity score analyses both the strength of expression and the inhomogeneity of expression. Figure 1: Distribution of the IRS across the cohort Figure 2: Overall Survival depending on the IRS Figure 3: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Table 1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	2	1	Based on the cell line RNA and protein data the expression level of MACC1 is highly variable rather than being present or not. Is there a similar variability present among the tissue samples? Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?	1	2	cancers14071773_makarova	1
+We thank the reviewer for raising this point. However, to respond to this question appropriately, we currently can provide only the results from proliferation assays for one cell line model (FLO1-EV vs FLO1-MACC1) over the time period of 72 h. This assay was with no statistically significant difference in cell proliferation between FLO1-EV vs FLO1-MACC1 (see Figure 4). Figure 4: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay. Unfortunately, the short revision period of 10 days limits the generation of additional data sets for proliferation in further tumor cell lines.	2	1	The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models. However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours. In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.	1	2	cancers14071773_makarova	1
+We thank the reviewer for this question. The concept of MACC1 inhibition was originally developed by our group for the treatment of MACC1 positive colorectal cancers and was recently published (Kobelt et al. Oncogene 2021). In this study we transferred our experience with MACC1 inhibition by selumetinib on AGE/S. Therefore, we chose the same doses for in vitro and in vivo experiments, and which was also used by other authors (Huynh et al. Mol Cancer Therapy. 2007). Further, we knew that the selected selumetinib concentrations of 10 µM has no cytotoxic effect in vitro and showed no reduction of migration in the MACC1 negative clones (FLO1-EV and OEshMACC1). Regarding the different dosages in the context of xenografted mice treatment in vivo and the clinically used dosages, the FDA recommends multiplying the dosage by a factor of 0.08 when transferring from mouse to human (Niar et al. Journal of basic and clinical pharmacy 2016). According to this calculation, the dosage of 50 mg selumetinib /kg mouse used would equal a dosage of 4 mg/kg in humans. The clinical dose of 75mg twice daily corresponds to a dose of 2x1mg per kg assuming a patient weight of 75kg. Using the aforementioned calculation, this dose equals then 12.5 mg/kg for treatment of mice. In our study, we refer again to the work of Huynh et al. Here, in which a dosage of 12 mg/kg selumetinib showed only a low level of effectiveness. At 100 mg/kg, a very good effectiveness was seen, however combined with an increased toxicity. Thus, we used in our study the dosage of 50 mg/kg as an effective dosage with low toxicity for the mice. Overall, we are considering here a pre-clinical model. The possibility of MEK1 inhibition to inhibit MACC1-induced effects in AGE/S was examined. Further, extended in vivo studies might then reveal the ideal dosage in vivo, being thereafter translated to use in humans. The use of trametinib is an interesting suggestion and might well be adapted to its use in forthcoming studies References: Kobelt, D., Perez-Hernandez, D., Fleuter, C., Dahlmann, M., Zincke, F., Smith, J., ... & Stein, U. (2021). The newly identified MEK1 tyrosine phosphorylation target MACC1 is druggable by approved MEK1 inhibitors to restrict colorectal cancer metastasis. Oncogene, 40(34), 5286-5301. Huynh H, Soo KC, Chow PK, Tran E. Targeted inhibition of the extracellular signal-regulated kinase kinase pathway with AZD6244 (ARRY-142886) in the treatment of hepatocellular carcinoma. Mol Cancer Ther. 2007 Jan;6(1):138-46.. Nair, Anroop B.; Jacob, Shery. (2016). A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy, 7. Jg., Nr. 2, S. 27.	2	1	It would be nice to know the selumetinib sensitivity of the two used cell lines. The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50mg/kg) are quite high. Please discuss how the applied concentrations are related to the clinicaly used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.	1	2	cancers14071773_makarova	1
+We thank the reviewer for this comment. The IRS cut-off value 5 reaches a sensitivity of 76.6% and a specificity of 64.2% (Youden Index. We include the arguments into the methods section page 3: To define the cut-off value a ROC curve analysis was performed. 0.408) and was chose as cut-off value (see supplement Figure S1 and Table S1). Additionally, we added the ROC curve and the table with sensitivity, specificity and Youden Index into a supplement section as figure S1 and table S1 Figure S1: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Positive if Greater than or Equal to Sensitivity 1- Specificity Specificity Youden Index -1 1 1 0 0 1 0,869 0,500 0,500 0,369 3 0,864 0,483 0,517 0,381 5 0,766 0,358 0,642 0,408 7 0,729 0,342 0,658 0,387 8,5 0,421 0,217 0,783 0,204 10,5 0,411 0,200 0,800 0,211 13 0 0 1 0 Table S1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	4	1	The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.	3	2	cancers14071773_makarova	1
+thank you for this comment: we added the proliferation analyses by MTT into the method section page 5: Additionally, proliferation for FLO-1/EV and FLO-1/MACC1 was analyzed over 72 h by MTT. 4x103 cells were plated into 96-well-plates and were allowed to accommodate for 24 h. after 48 and 72 h formazan crystals were dissolved in 150 μl of DMSO and the absorption was measured at 560 nm in the absorbance reader (Tecan infinite 200 PRO). Each cell proliferation experiment was performed in triplicates. We added the results of the MTT analyses into the results section page 11: The additional analysis of the proliferation over 72 h showed no significant differences between FLO-1/EV (100% ± 4.886) and FLO-1/MACC1 (112.3% ± 12.24; p=0.113) (see supplement Figure S3). We added the MTT plot as figure S2 into the supplement section Figure S2: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay.	4	1	"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."	3	2	cancers14071773_makarova	1
+We apologize for the inconvenience. Figure 4 can be found on page 14 in the submitted version. We inserted Figure 4 in the revised manuscript again to solve the technical issue.	2	1	[1] Figure 4 is missing. In the pdf I reviewed, the caption is there, but the figure is not.	1	2	cancers14071773_makarova	1
+We address this question in the results part MACC1 expression: “ Due to the procedure of cutting and staining, some samples could not be used for the evaluation. Samples with insufficiently representative tumor tissue were also excluded from the evaluation. Cores with representative tumor material and evaluable staining were available in 266 of 360 samples (73.9%)…”. To avoid misunderstandings, we corrected the abstract to: 266 Samples of 360 AGE/S patients were analyzed for MACC1 expression.	2	1	[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249). Please fix this inconsistency.	1	2	cancers14071773_makarova	1
+We thank the reviewer for this correction. This formatting issue was corrected throughout the entire manuscript.	2	1	"[3] Throughout the paper, 10^x wrongly displays as 10x (e.g. ""105"" instead of ""10^5"" in lines 152 and 170, ""106"" instead of ""10^6"" in line 217, ""X2"" instead of ""X^2"" in line 241, ""108"" instead of ""10^8"" in lines 353, 354, 365, 366, 367, 368, 370, 371)."	1	2	cancers14071773_makarova	1
+We thank the reviewer for this valuable hint. As suggested, we included p-values in Figure 3G and 3H. In Figure 3G there is a significant increase in migration of cells without MACC1 expression. This is based on a sufficiently large N, that renders even small differences statistically significant. Further, this increase is rather small and is outperformed by the MACC1-mediated increase in migration and the reduction in the MACC1-mediated migration by selumetinib. The data show, that selumetinib does not reduce migration of the MACC1-negative FLO1 and that MACC1 induces migration in this cell line that can be inhibited by selumetinib.	2	1	[4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively. If the difference is statistically significant, please briefly explain/speculate why.	1	2	cancers14071773_makarova	1
+We thank the reviewer for this suggestion to improve the visual quality of the manuscript. The legends of 3F and I have been enlarged and the resolution of the figure has been increased.	2	1	[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).	1	2	cancers14071773_makarova	1
+We thank the reviewer for this discussion. The empty vector control clone was transduced and selected afterwards. During the selection procedure it is possible that a subclone was generated that expresses slightly more (or less) of any mRNA and protein. It differs from the wildtype by handling (transduction, selection etc) and somewhat by age (cell divisions). Therefore, for all comparisons the empty vector clone was used, as this clone was generated in parallel to the MACC1 clone.	2	1	[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.	1	2	cancers14071773_makarova	1
+We thank the reviewer for pointing this out: Survival reports were changed in median survival and the 95% confidence interval were included. Additionally, disease-specific survival results were reported.	2	1	[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).	1	2	cancers14071773_makarova	1
+We thank the reviewer for pointing this out: OR with the confidence interval is now included.	2	1	"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e. specify the 95% confidence interval."	1	2	cancers14071773_makarova	1
+We thank the reviewer for pointing this out: all survival reports were changed in median survival and the 95% confidence interval were included.	2	1	[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values). Same for lines 425/426.	1	2	cancers14071773_makarova	1
+Thank you for this valuable comment. The journal requires an unstructured abstract. As suggested by the reviewer, we have therefore reorganized the abstract and improved the flow of reading: Abstract: Esophageal and Gastric Adenocarcinomas (AGE/S) are characterized by early metastasis and poor survival. MACC1 (Metastasis Associated in Colon Cancer 1) acts in colon cancer as a metastasis inducer, linked to reduced survival. This project illuminates the role and potential of inhibition of MACC1 in AGE/S. Using 266 of 360 TMAs and survival data of AGE/S patients we confirm the value of MACC1 as an independent negative prognostic marker in AGE/S patients. MACC1 gene expression correlated with survival and morphological characteristics. In vitro analysis of lentivirally MACC1 manipulated subclones of FLO-1 and OE33 showed enhanced migration induced by MACC1 in both cell line models, which could be inhibited by the MEK1 inhibitor selumetinib. In vivo, the efficacy of selumetinib on tumor growth and metastasis of MACC1-overexpressing FLO-1 cells xenografted intrasplenically in NOG mice was tested. Mice with high MACC1 expressing cells developed faster and larger distant metastases. Treatment with selumetinib led to a significant reduction of metastasis exclusively in the MACC1 positive xenografts.	2	1	"[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion. If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g. ""MACC1 is an independent negative prognostic marker in AGE/S patients."" should be after ""Expression was correlated with survival and morphological characteristics."" and before ""To analyze the role of MACC1 in vitro the cell lines FLO-1 and OE33 were lentivirally manipulated."" and so on)."	1	2	cancers14071773_makarova	1
+There were no more open questions in the second review.	4	1	The authors have satisfactorily addressed my comments.	3	2	cancers14071773_makarova	1
+As suggested, we analyzed the distribution of IRS (immunoreactivity score) across the cohort. In Figure 1 it can be clearly seen that there are two groups forming here, those with 0-4 and those with 6-12 IRS. In addition, we calculated the survival probability depending on the IRS (Figure 2). Patients with an IRS of 6 or higher have a similar bad prognosis compared to patients with an IRS below 5. A better survival correlates with lower MACC1 IRS. Patients with an IRS of 0 have the best prognosis. Based on these two analyses, the decision for cut-off value of 5 was made. Additionally, ROC curve analysis showed, that the IRS value of 5 has the highest Youden Index and is therefore the most suitable cut-off value. 0.408) (Figure 3 and Table 1). Looking at MACC1 expression in cell lines and tissue samples, the variability looks similar. Nevertheless, it must be considered that the investigation of the homogeneous cell lines is a purely quantitative analysis of the expression by Western plot and RT-PCR. The analysis of the tissue samples by immunoreactivity score analyses both the strength of expression and the inhomogeneity of expression. Figure 1: Distribution of the IRS across the cohort Figure 2: Overall Survival depending on the IRS Figure 3: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Table 1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	2	1	Based on the cell line RNA and protein data the expression level of MACC1 is highly variable rather than being present or not. Is there a similar variability present among the tissue samples? Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?	1	2	cancers14071773_perova	1
+We thank the reviewer for raising this point. However, to respond to this question appropriately, we currently can provide only the results from proliferation assays for one cell line model (FLO1-EV vs FLO1-MACC1) over the time period of 72 h. This assay was with no statistically significant difference in cell proliferation between FLO1-EV vs FLO1-MACC1 (see Figure 4). Figure 4: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay. Unfortunately, the short revision period of 10 days limits the generation of additional data sets for proliferation in further tumor cell lines.	2	1	The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models. However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours. In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.	1	2	cancers14071773_perova	1
+We thank the reviewer for this question. The concept of MACC1 inhibition was originally developed by our group for the treatment of MACC1 positive colorectal cancers and was recently published (Kobelt et al. Oncogene 2021). In this study we transferred our experience with MACC1 inhibition by selumetinib on AGE/S. Therefore, we chose the same doses for in vitro and in vivo experiments, and which was also used by other authors (Huynh et al. Mol Cancer Therapy. 2007). Further, we knew that the selected selumetinib concentrations of 10 µM has no cytotoxic effect in vitro and showed no reduction of migration in the MACC1 negative clones (FLO1-EV and OEshMACC1). Regarding the different dosages in the context of xenografted mice treatment in vivo and the clinically used dosages, the FDA recommends multiplying the dosage by a factor of 0.08 when transferring from mouse to human (Niar et al. Journal of basic and clinical pharmacy 2016). According to this calculation, the dosage of 50 mg selumetinib /kg mouse used would equal a dosage of 4 mg/kg in humans. The clinical dose of 75mg twice daily corresponds to a dose of 2x1mg per kg assuming a patient weight of 75kg. Using the aforementioned calculation, this dose equals then 12.5 mg/kg for treatment of mice. In our study, we refer again to the work of Huynh et al. Here, in which a dosage of 12 mg/kg selumetinib showed only a low level of effectiveness. At 100 mg/kg, a very good effectiveness was seen, however combined with an increased toxicity. Thus, we used in our study the dosage of 50 mg/kg as an effective dosage with low toxicity for the mice. Overall, we are considering here a pre-clinical model. The possibility of MEK1 inhibition to inhibit MACC1-induced effects in AGE/S was examined. Further, extended in vivo studies might then reveal the ideal dosage in vivo, being thereafter translated to use in humans. The use of trametinib is an interesting suggestion and might well be adapted to its use in forthcoming studies References: Kobelt, D., Perez-Hernandez, D., Fleuter, C., Dahlmann, M., Zincke, F., Smith, J., ... & Stein, U. (2021). The newly identified MEK1 tyrosine phosphorylation target MACC1 is druggable by approved MEK1 inhibitors to restrict colorectal cancer metastasis. Oncogene, 40(34), 5286-5301. Huynh H, Soo KC, Chow PK, Tran E. Targeted inhibition of the extracellular signal-regulated kinase kinase pathway with AZD6244 (ARRY-142886) in the treatment of hepatocellular carcinoma. Mol Cancer Ther. 2007 Jan;6(1):138-46.. Nair, Anroop B.; Jacob, Shery. (2016). A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy, 7. Jg., Nr. 2, S. 27.	2	1	It would be nice to know the selumetinib sensitivity of the two used cell lines. The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50mg/kg) are quite high. Please discuss how the applied concentrations are related to the clinicaly used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.	1	2	cancers14071773_perova	1
+We thank the reviewer for this comment. The IRS cut-off value 5 reaches a sensitivity of 76.6% and a specificity of 64.2% (Youden Index. We include the arguments into the methods section page 3: To define the cut-off value a ROC curve analysis was performed. 0.408) and was chose as cut-off value (see supplement Figure S1 and Table S1). Additionally, we added the ROC curve and the table with sensitivity, specificity and Youden Index into a supplement section as figure S1 and table S1 Figure S1: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Positive if Greater than or Equal to Sensitivity 1- Specificity Specificity Youden Index -1 1 1 0 0 1 0,869 0,500 0,500 0,369 3 0,864 0,483 0,517 0,381 5 0,766 0,358 0,642 0,408 7 0,729 0,342 0,658 0,387 8,5 0,421 0,217 0,783 0,204 10,5 0,411 0,200 0,800 0,211 13 0 0 1 0 Table S1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	4	1	The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.	3	2	cancers14071773_perova	1
+thank you for this comment: we added the proliferation analyses by MTT into the method section page 5: Additionally, proliferation for FLO-1/EV and FLO-1/MACC1 was analyzed over 72 h by MTT. 4x103 cells were plated into 96-well-plates and were allowed to accommodate for 24 h. after 48 and 72 h formazan crystals were dissolved in 150 μl of DMSO and the absorption was measured at 560 nm in the absorbance reader (Tecan infinite 200 PRO). Each cell proliferation experiment was performed in triplicates. We added the results of the MTT analyses into the results section page 11: The additional analysis of the proliferation over 72 h showed no significant differences between FLO-1/EV (100% ± 4.886) and FLO-1/MACC1 (112.3% ± 12.24; p=0.113) (see supplement Figure S3). We added the MTT plot as figure S2 into the supplement section Figure S2: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay.	4	1	"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."	3	2	cancers14071773_perova	1
+We apologize for the inconvenience. Figure 4 can be found on page 14 in the submitted version. We inserted Figure 4 in the revised manuscript again to solve the technical issue.	2	1	[1] Figure 4 is missing. In the pdf I reviewed, the caption is there, but the figure is not.	1	2	cancers14071773_perova	1
+We address this question in the results part MACC1 expression: “ Due to the procedure of cutting and staining, some samples could not be used for the evaluation. Samples with insufficiently representative tumor tissue were also excluded from the evaluation. Cores with representative tumor material and evaluable staining were available in 266 of 360 samples (73.9%)…”. To avoid misunderstandings, we corrected the abstract to: 266 Samples of 360 AGE/S patients were analyzed for MACC1 expression.	2	1	[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249). Please fix this inconsistency.	1	2	cancers14071773_perova	1
+We thank the reviewer for this correction. This formatting issue was corrected throughout the entire manuscript.	2	1	"[3] Throughout the paper, 10^x wrongly displays as 10x (e.g. ""105"" instead of ""10^5"" in lines 152 and 170, ""106"" instead of ""10^6"" in line 217, ""X2"" instead of ""X^2"" in line 241, ""108"" instead of ""10^8"" in lines 353, 354, 365, 366, 367, 368, 370, 371)."	1	2	cancers14071773_perova	1
+We thank the reviewer for this valuable hint. As suggested, we included p-values in Figure 3G and 3H. In Figure 3G there is a significant increase in migration of cells without MACC1 expression. This is based on a sufficiently large N, that renders even small differences statistically significant. Further, this increase is rather small and is outperformed by the MACC1-mediated increase in migration and the reduction in the MACC1-mediated migration by selumetinib. The data show, that selumetinib does not reduce migration of the MACC1-negative FLO1 and that MACC1 induces migration in this cell line that can be inhibited by selumetinib.	2	1	[4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively. If the difference is statistically significant, please briefly explain/speculate why.	1	2	cancers14071773_perova	1
+We thank the reviewer for this suggestion to improve the visual quality of the manuscript. The legends of 3F and I have been enlarged and the resolution of the figure has been increased.	2	1	[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).	1	2	cancers14071773_perova	1
+We thank the reviewer for this discussion. The empty vector control clone was transduced and selected afterwards. During the selection procedure it is possible that a subclone was generated that expresses slightly more (or less) of any mRNA and protein. It differs from the wildtype by handling (transduction, selection etc) and somewhat by age (cell divisions). Therefore, for all comparisons the empty vector clone was used, as this clone was generated in parallel to the MACC1 clone.	2	1	[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.	1	2	cancers14071773_perova	1
+We thank the reviewer for pointing this out: Survival reports were changed in median survival and the 95% confidence interval were included. Additionally, disease-specific survival results were reported.	2	1	[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).	1	2	cancers14071773_perova	1
+We thank the reviewer for pointing this out: OR with the confidence interval is now included.	2	1	"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e. specify the 95% confidence interval."	1	2	cancers14071773_perova	1
+We thank the reviewer for pointing this out: all survival reports were changed in median survival and the 95% confidence interval were included.	2	1	[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values). Same for lines 425/426.	1	2	cancers14071773_perova	1
+Thank you for this valuable comment. The journal requires an unstructured abstract. As suggested by the reviewer, we have therefore reorganized the abstract and improved the flow of reading: Abstract: Esophageal and Gastric Adenocarcinomas (AGE/S) are characterized by early metastasis and poor survival. MACC1 (Metastasis Associated in Colon Cancer 1) acts in colon cancer as a metastasis inducer, linked to reduced survival. This project illuminates the role and potential of inhibition of MACC1 in AGE/S. Using 266 of 360 TMAs and survival data of AGE/S patients we confirm the value of MACC1 as an independent negative prognostic marker in AGE/S patients. MACC1 gene expression correlated with survival and morphological characteristics. In vitro analysis of lentivirally MACC1 manipulated subclones of FLO-1 and OE33 showed enhanced migration induced by MACC1 in both cell line models, which could be inhibited by the MEK1 inhibitor selumetinib. In vivo, the efficacy of selumetinib on tumor growth and metastasis of MACC1-overexpressing FLO-1 cells xenografted intrasplenically in NOG mice was tested. Mice with high MACC1 expressing cells developed faster and larger distant metastases. Treatment with selumetinib led to a significant reduction of metastasis exclusively in the MACC1 positive xenografts.	2	1	"[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion. If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g. ""MACC1 is an independent negative prognostic marker in AGE/S patients."" should be after ""Expression was correlated with survival and morphological characteristics."" and before ""To analyze the role of MACC1 in vitro the cell lines FLO-1 and OE33 were lentivirally manipulated."" and so on)."	1	2	cancers14071773_perova	1
+There were no more open questions in the second review.	4	1	The authors have satisfactorily addressed my comments.	3	2	cancers14071773_perova	1
+We thank the reviewer for this important question and would be happy to clarify. TTFields are alternating electric fields in the range of 100 to 500 kHz, with maximal efficacy seen at a different frequency for different tumor types. Since TTFields cannot be applied at several different frequencies simultaneously, one specific frequency needs to be selected, and so the first step in examination of a new tumor type is a frequency scan. The effect of TTFields is not dichotomic, such that only one frequency is effective while the other are not at all. From the frequencies showing efficacy the purpose is to select the most effective one. It should be clarified that higher frequencies of the alternating electric fields do not mean higher energy, and hence there is no disadvantage in working with higher frequencies and no added toxicity or increased side effects for higher frequencies within the TTFields frequency range. Of note, TTFields at 200 kHz is already approved and has been applied to more than 18,000 patients with glioblastoma, with skin irritation being the main treatment related adverse event. TTFields at 150 kHz is approved for treatment of unresectable malignant pleural mesothelioma. To address these issues, we have rephrased the discussion: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” and “The lower TTFields intensity required for treatment of HepG2 relative to Huh-7D12 cells for achieving the same level of efficacy suggest higher sensitivity of the former to TTFields.” Point 2: What is the ‘n’ number of the samples and experiments in each group?	2	1	What is the rationale of using 150 kHz TTFields? TTFields at 100 kHz has also shown significant differences as cytotoxicity at p<0.01 and p<0.001 in HepG2 and Huh-7D12 cells which have around 60% and 50% of cell survival. Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).	1	2	cancers14122959_makarova	1
+We thank the reviewer for pointing this was not clear. As mentioned in the statistical analysis sub-section within the methods part, all in vitro experiments were repeated at least 3 times, and so depicted values are mean (N ≥ 3) ± SEM. We have now added this also to the legends of figures 1, 2, and 3. Per the in vivo study, as was mentioned in the methods (sub-section 2.8), 52 animals were included in the study. The specific numbers in each treatment group are now mentioned in the results (sub-section 3.4): “During the treatment period, average tumor volumes of control animals (n = 11) increased 5.9-fold (Figure 4b and 4c; and Figure S3 for tumor images). For animals treated with TTFields (n = 15) or sorafenib (n = 10), tumor growth was significantly lower, 3.3- and 2.3-fold, respectively. In the TTFields-sorafenib combination group (n = 16), a 1.6-fold increase in tumor volume within the treatment period was observed, a growth significantly lower than that for control or for each treatment alone.” It is also mentioned in the legend of figure 4: “Rats (n = 52) were inoculated orthotopically with rat HCC N1S1 cells, and treated with sham-vehicle (n = 11), TTFields alone (n = 15), sorafenib alone (n = 10), or TTFields + sorafenib (n = 16), according to the depicted timeline (a).” Point 3: LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group.	2	1	What is the ‘n’ number of the samples and experiments in each group? Also mention the ‘n’ number in all the experiments involved for invitro and invivo.	1	2	cancers14122959_makarova	1
+First, we would like to emphasize that indications of autophagy were diverse and determined from increased cellular granularity, amplified lipidation of LC3 (from LC3-I to LC3-II) detected by Western blot, and elevated levels of LC3 foci observed by fluorescent staining. Nevertheless, to better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and additional time points. As per the reviewer’s request, these examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3.	2	1	LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group. How can the authors correlate autophagy with apoptosis? It is not so trustworthy that the only expression of LC3B indicate the treated condition have increased autophagy in the tumor tissue. Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.	1	2	cancers14122959_makarova	1
+Per the reviewer’s request we show below images of the tumors within the liver, which we have also provided now in the supporting material of the paper. As may be seen, the liver itself is much larger than the tumor, and so we did not see value in measuring total liver weight, and hence cannot provide these values. While we tried to reduce variability between the animals by excluding those that at treatment initiation had tumor volumes not within the range 30–100 mm3, there was still divergence between the animals that resulted in the standard deviation bars depicted in the figure. It is however important to mention that there was a very good correlation between tumor volume (as measured by MRI) versus tumor weight at the end of the study, as may be seen in the graph below.	2	1	Several data have shown very high error bars in each group (especially Fig Can the author provide the tumor images which were harvested from mice?	1	2	cancers14122959_makarova	1
+IHC tests the presence of a protein already expressed in the tissue, and 30 min are sufficient to allow recognition and binding of the antibody to the target protein. The secondary antibody is an HRP conjugated goat anti rabbit, that is a part of the Leica HRP-refine detection kit (Cat # DS9800). We have added the missing antibody information to section 2.10. We unfortunately do not have any tissue left to allow for performing WB analysis from the in vivo study, but we have conducted additional cell line studies and added WB analysis of cleaved PARP for the in vitro examinations and additional IHC examinations for autophagy and ER stress markers (as shown in response 3).	2	1	In the IHC experiment for PARP, the authors have incubated with primary antibody only for 30 min? Is that timing enough to get protein expression? What source of secondary antibody was used? In addition to IHC, I suggest performing western blot using PARP antibody where the full length and cleaved bands are observed in the same blot.	1	2	cancers14122959_makarova	1
+We thank the reviewer for identifying this mistake, and have corrected this, changing the label of the LC3 data to e.	2	1	The figure number is mislabeled in Fig 4D-LC3 Immunofluorescence.	1	2	cancers14122959_makarova	1
+Error bars are not missing for the control groups, rather they are null, as the values displayed in these figures are relative to control, and so per definition all control experiments have the exact same value of 100 or 1 (for percentage or fold change, respectively).	2	1	Why there is no error bar in the control group of all bar graphs? Please include error bars and re-calculate the statistical analysis for all the data wherever missing.	1	2	cancers14122959_makarova	1
+We apologize for this mistake, and have now corrected the figure legend. The quantification of immunofluorescence of LC3 foci formation is c, and immunoblotting showing LC3-II to LC3-I ratio is d. Point 9: There is no data in Fig 4A.	2	1	Figure legends of Fig2D is missing. Fig C is repeated in the legend. Please label the figures appropriately. It is so frustrating to understand.	1	2	cancers14122959_makarova	1
+We apologize for accidently omitting Fig 4A. This figure depicts the timeline of the in vivo experiment for easier understanding of the study designed. We have now corrected this.	2	1	There is no data in Fig 4A. Whole data is missing but have explained in the result section and in figure legends.??	1	2	cancers14122959_makarova	1
+We understand from the reviewer’s comments that the sentence “The higher effects seen in the presence of CQ reveal that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation” is not clear enough. We have hence rephrased the sentence to be more accurate and better deliver the message: “CQ is an inhibitor of lysosome degradation, commonly used to decipher whether the elevation of LC3 is due to upregulation of the autophagy process or reduced autophagosome turnover [30]. The higher TTFields-induced elevation of LC3 seen in the presence of CQ suggest that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation.” Point 11: Typos: Line 111, ‘invitro’ spelling is not correct.	2	1	Line 337 seems over statement since the data are not shown in the manuscript.	1	2	cancers14122959_makarova	1
+The “inovitro” in line 111 is not a typo, it is the name of the system used for applying TTFields in vitro. In line 342 we removed the duplicate “and”, and thank the reviewer for catching this typo.	2	1	Typos: Line 111, ‘invitro’ spelling is not correct. Double ‘and’ in Line 342. Need language and grammar check.	1	2	cancers14122959_makarova	1
+We thank the reviewer for the positive review.	2	1	This is an interesting scientific study, as it concerns the issue of hepatocellular carcinoma, it also has a clinical aspect. The materials and methods section are elaborated in the details. Therefore, I expect further additional examination in the future. References contain mostly publications printed in the last 10 years. The authors indicated that TTFields had potential to be a new treatment option of hepatocellular carcinoma. The present study was examined in terms of autophagy and apoptosis in the antitumor effects of TTFields and sorafenib. However, the exact mechanism of the combination therapy induced cell death is not yet known as the activation of autophagy in the combination therapy was not increased.	1	2	cancers14122959_makarova	1
+For the in vitro experiments with TTFields (sub-section 2.3) we tried not to elaborate too much and referenced previous work describing all details. We understand that we have cut off too many details, and are happy to add them “HepG2 and Huh-7D12 cell suspensions (500 µl, 25 x 103 cells/plate) were placed as a drop in the center of 35-mm inovitro™ dishes composed of high dielectric constant ceramic (lead magnesium niobate–lead titanate [PMN-PT]), with two perpendicularly pairs of transducer arrays printed on their outer walls. Cells were incubated overnight at 37 °C to allow attachment to the dish, and then 2 ml of fresh media were added.” Regarding the number of rats included in the final analysis, we apologize this information was not clear. The 52 rats mentioned in the text were in fact the actual final number of animals in the analysis, since “all rats reached the required usage limit of ≥18 h/day”, as is now explicitly stated in the results sub-section 3.4.” Point 2: What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells.	2	1	In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells). A very important missing is not showing the actual number of rats included in the final analyses (the ones who successfully received therapy for more than 18 hours/day). This must be added.	1	2	cancers14122959_makarova	1
+We thank the reviewer for raising this important issue. Using a xerograph model is indeed a good suggestion, however it would require working in immunosuppressed mice. It is currently not feasible for us to perform studies in which we apply TTFields to immunosuppressed animals, as these animals are of smaller weight thus the burden of wearing the electrodes may induce too much stress. Furthermore, these animals are more prone to contract infections during the process of electrodes placement and replacement on the animals. On the other hand, using the rat cells for in vitro experiments in the inovitro dishes turned out to be very technically challenging, as the cells were non-adherent, and so we could not pursue this important avenue. Nevertheless, we agree with the reviewer that there is a possibility that the optimal frequency may differ in the N1S1 relative to the human cell lines. Therefore, we have addressed this issue by including results of an additional frequency scan experiments done with N1S1 cells. We now mention this in the text, results sub-section 3.4: “The efficacy of combining TTFields with sorafenib relative to each modality alone was examined in the N1S1 HCC rat orthotopic model (timeline in Figure 4a). In vitro experiments confirmed that the 150 kHz TTFields frequency found optimal for treatment of the human cell lines was also optimal for treatment of the murine N1S1 cells used for the in vivo study (Figure S2).” We provide the relevant frequency scan figure in the supplementary material. Per the good question by the reviewer regarding the p53 status of the N1S1 cells we added: “It is also worth mentioning that the N1S1 murine cells used for this study, like the HepG2 cells, are p53 wild type.” Figure S2.	2	1	What are the p53 status and the apoptosis signaling pathway function in N1S1 cells? Maybe the cytotoxicity data will reveal a different better frequency.	1	2	cancers14122959_makarova	1
+We thank the reviewer for this comment, and apologize that the illustration of the timeline was accidentally missing from the submission (we have now added it). The limiting factor for study duration was the well being of the animals. The tumors in the control group were very large, causing stress and weight loss of the animals. The arrays placed on the animals, together with the individual housing needed to prevent wire entanglement, adds even more stress and increases animal weight loss. Overall, it was non-ethical to continue the study further. We have now added this explanation to the discussion section to clarify this limitation of the study: “In the HCC animal model, the acute effects of TTFields and sorafenib were examined. Due to the large tumors developed in the control group and the stress experienced by the animals as a result of the individual housing and motility limitations imposed by the sham and TTFields arrays, longer treatment durations were not feasible.” Point 4: There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing.	2	1	Another puzzling experiment is the schedule for the in vivo work. Since the authors missed to add Figure 4A for timeline, based on Methods section the rats were treated for 5 days with TTFields and or sorafenib and a day later the rays were sacrificed. While a short “acute” follow-up is welcome the most important experiment should allow the follow-up for much more days to indeed observe the effect of TTFields added to sorafenib. I could not find an explanation for not including a long term follow-up. In my opinion, this is a key therapeutically experiment which must be done and included in the study.	1	2	cancers14122959_makarova	1
+It is important to understand that the TTFields frequency is not the treatment dose, and hence increased frequency does not mean increased efficacy. In the case of TTFields, intensity, duration, and usage are the factors that contribute to the “treatment dose”. Nevertheless, we have added to the discussion an explanation on this issue of optimal frequency: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” Regarding the option that the optimal frequency will differ between in vitro and in vivo setting, this is indeed a concern. However, as shown by the in vitro frequency scans, the effect of TTFields is not dichotomic, and effectiveness can be seen at more than one frequency. Therefore, the relatively low expected differences between animals treated with different TTFields frequencies together with the variability inherent to animal studies, makes it technically problematic to perform frequency scans in vivo. As of today, in vitro frequency scans are the common practice for determining the optimal TTFields frequency for delivery to animals and to humans. In fact, in vitro frequency scans were the basis for the clinical studies leading to FDA approval of TTFields at 200 kHz for the treatment of GBM and at 150 kHz for the treatment of MPM. We have added this issue to the discussion as a limitation of the study: “Since the optimal TTFields frequency has been shown to be dependent on the electrical properties of the cells and it is not clear how much effect the tumor microenvironment has on these properties, and because it is technically problematic to perform TTFields frequency scans in vivo, the frequency detected in the cell cultures was also employed for the animal studies.” Point 5: Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	2	1	There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing. Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is total different?	1	2	cancers14122959_makarova	1
+When it comes to testing efficacy, we would like to use a treatment duration as long as possible to maximize the effect. However, this duration varies between in vitro and in vivo work. For cell cultures, the control samples continue to grow throughout the treatment duration, and so after 72 h the plates are very much confluent, and may not be left to continue and grow any further (without drastic changes in the environmental conditions, making them inadequate control cells). It is also noteworthy that such treatment periods for in vitro work are well accepted in the field of TTFields research. In vivo, prior studies also used similar time frames of 2 to 3 weeks from inoculation when working with the N1S1 model (Buijs et al., 2012 -; Ju et al., 2009; Thompson et al., 2012). Extending treatment duration was also limited by the physical status of the animals, as was explained in response 3.	2	1	Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	1	2	cancers14122959_makarova	1
+Chloroquine was added to the cell cultures only at the final hours of the treatment for answering questions related to the mechanism of action, and not for boosting the efficacy of the other treatments. When using chloroquine in animal studies, it is for efficacy purposes, and so it is used throughout the treatment period. Since our animal experiment aimed to examine the efficacy of concomitant TTFields and sorafenib, with no additional agents, chloroquine was not employed.	2	1	Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.	1	2	cancers14122959_makarova	1
+For the cytotoxicity assay we remove the supernatant, washed the cells, and then collect the adherent cells following trypsinization and visual inspection to verify all cells were removed. Indeed, as the reviewer mentioned correctly, we “counted the live cells and plotted the final data as percentage of untreated controls”. We agree that this is not cytotoxicity per se, and that is why we also perform 7-AAD/annexin-V staining of the cells. For this apoptosis assay we do collect the supernatant together with the adherent cells. Increased apoptosis and/or necrosis indicates that reduction in the cell number observed in the cell count emanates at least in part from cytotoxicity. We agree with the reviewer that we should not confuse the cell count measurements with the term “cytotoxicity” prior to showing the effect on apoptosis, and therefore we have changed this terminology throughout the paper.	2	1	The authors claim that cytotoxicity was measured “by cell counting using iCyt EC800 (Sony Biotechnology) 123 flow cytometer, and expressed as a percentage relative to the control.” Does this imply that they counted the live cells and plotted the final data as percentage of untreated controls (as figure 1 suggests). However, is this a real cytotoxicity or a cell growth inhibition? Did they measure the adherent cells after trypsinization. Were the cells from supernatant counted (where are probably the majority of dead cells)? There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.	1	2	cancers14122959_makarova	1
+We thank the reviewer for these questions. Preliminary tests using CD31 staining revealed no differences between the groups regarding blood vessel density and therefore we did not pursue the research in that direction. We agree that measuring the effect of TTFields on the anti-angiogenic effect of sorafenib and on resistance to sorafenib are interesting, these topics were however not within the scope of the current study, and remain for future investigations. Regarding possible discrepancy between the volume fold change and tumor weight, please see below a graph showing tumor volume (as measured by MRI) versus tumor weight, both measured at the end of the study. The graph shows very good correlation between the two parameters, indicating the reliability of the measurements. If the reviewer feels there is discrepancy, it may be due to the volume shown as fold change relative to the initial tumor volume and not as the end value.	2	1	Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated? Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic? Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group. Did the authors check changes in blood vessels density. Were the mice perfused before collecting the tumors?	1	2	cancers14122959_makarova	1
+We routinely examine our cells for Mycoplasma. Regarding authenticity, the cells were used shortly after purchase, and so there was no need to examine this.	2	1	Was Mycoplasma testing done routinely? Were the cells checked also for authenticity?	1	2	cancers14122959_makarova	1
+We appreciate this question. Autophagy is a process that is elevated in order to cope with cellular stress, but as stress level elevate, autophagy can no longer provide the protection the cell needs, and the cell will undergo apoptosis. This kind of kinetics means that the levels of autophagy markers will depend on the time point the cells are examined. This may be appreciated from figure 2 panel c and d, with the different kinetics displayed by the two cell lines. While for Huh-7D12 autophagy levels increase from 24 to 48 hours of treatment, in the HepG2 cells autophagy levels at 48 hours are lower than at 24 hours, indicating these cells are already after the autophagy peak and on the way to apoptosis. Indeed, figure 1d shows higher levels of apoptosis for HepG2 cells. To clarify we have added a few sentences to the text. In results sub-section 3.2: “However, autophagy kinetics seems to be faster in the HepG2 cells, in which LC3 markers are lower at 48 versus 24 hours, whereas elevation is seen from 24 to 48 hours for the Huh-7D12 cells.” In the discussion “While autophagy serves as a survival strategy of cells, when stress levels continue raising it may be over-activated and mediate cell death [9]. The faster autophagy kinetics seen for the HepG2 relative to Huh-7D12 cells following application of TTFields is in agreement with the higher apoptosis levels displayed by this cell line, and may serve as an additional rational for the higher efficacy of TTFields against it. Examination of the reasons for faster autophagy in HepG2 relative to Huh-7D12 cells is out of the scope of this work.” For more clarity we have also added a more in-depth kinetic study, including additional relevant markers and additional time points, examinations that were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case” Figure 3.	2	1	Is there any explanation why the combination of TTFields and sorafenib did not induce a significant level of autophagy as compared to untreated animals which invalid the initial hypothesis that “concomitant application of sorafenib and TTFields may increase stress levels enough to tilt autophagy towards the cell death pathway”.	1	2	cancers14122959_makarova	1
+The sentence referred to was meant to describe only the in vivo outcomes. We have rephrased it to be more accurate and clear: “While each treatment alone elevated levels of autophagy relative to control, TTFields concomitant with sorafenib induced a significant increase versus control in tumor ER stress and apoptosis levels, demonstrating increased stress under the multimodal treatment.” Point 12: Finally, adding to all the above questions, I found a very weak Discussion section which must be extended.	2	1	The statement “TTFields concomitant with sorafenib induced a significant increase in apoptosis’ in the abstract section is overstated. When compared with sorafenib alone there is practically no difference. Moreover, TTFields failed to increases apoptosis when added to sorafenib and compared to sorafenib alone in one out of two human cells line investigated	1	2	cancers14122959_makarova	1
+We thanks the reviewer for this comment. We have now elaborated on many issues throughout the discussion, as was described thorough this response letter. Regarding the conclusion, we have rephrased it for better accuracy: “TTFields were identified to be most efficient for treatment of HCC cells at 150 kHz, and this frequency further demonstrated in vivo efficacy.” Why only one frequency was used in vivo, and the difference between frequency and dose, were explained in response 4.	2	1	Finally, adding to all the above questions, I found a very weak Discussion section which must be extended. Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”. This is not a correct statement. While for in vitro data, the authors have data, for in vivo they used only one frequency of 150 Hz. At least one different dose should have been studied for comparison since this is a completely different tumor environment than the in vitro one.	1	2	cancers14122959_makarova	1
+We thank the reviewer for this question. Indeed, the sorafenib dose used in this study proved to be slightly more efficacious than TTFields in controlling tumor fold increase. Nevertheless, these differences between the monotherapies, did not reach statistical significance. In accordance, there was no statistical difference between the monotherapies in the expression levels of the LC3 marker and the levels of cleaved PARP. In order to better understand the mechanism of action, we have added experiments to better characterize the autophagy-apoptosis interplay for treatment with concomitant TTFields and sorafenib, which are described in response 3. Of note, the clinical development of TTFields does not aim to replace sorafenib with TTFields, but rather to add TTFields on top of sorafenib and therefore this work focused mainly on the potential added value in combining these 2 modalities.	2	1	1) The authors demonstrate the efficacy of TTFields in vivo even when used as monotherapy. As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib. However, no differences in expression of LC3 marker were observed between these groups (treated with TTFields or with sorafenib)(as shown in Figure 4D). Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F. What is the mechanism illustrating higher efficiency of sorafenib against HCC?	1	2	cancers14122959_makarova	1
+We thank the reviewer for pointing out this issue. Quantification of the IHC images was done automatically. The whole slide was scanned, and the CaseViewer software was used to exclude non-tumor areas. The signals of the stained protein and the nuclei were resolved by color deconvolution and quantified separately using the FIJI software (ImageJ) software. Average signal per cell or percent of positive cells was calculated. As the reviewer pointed out, the high magnification images we chose to show do not correctly reflect the quantification performed by the software, and we have now replaced them with better representative fields of the slides.	2	1	2) Despite the expression of cleaved PARP was very low in the tumors treated with TTFields or sorafenib alone ( as shown in IHC-images in Figure 4F), the authors declare about ~ 20% of positive cells, as show in the graphs below IHC-staining. Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.	1	2	cancers14122959_makarova	1
+We sincerely appreciate this well-taken comment. To better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and at additional time points. These examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3.	2	1	3) It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenin induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.	1	2	cancers14122959_makarova	1
+Both cell lines experience elevation of apoptosis following application of sorafenib in a dose dependent manner, as evident from the AnnV/7AAD results. However, while TTFields greatly elevate apoptosis in HepG2 cells, they have a low effect on apoptosis levels in the Huh-7D12 cells, seen both in Figure 1d and in Figure 3c. As was explained in the discussion, this difference between the cell lines may be attributed to the different p53 status, wild type in HepG2 and mutated in Huh-7D12, as there are previous indications of lower TTFields-induced apoptosis in cell lines with mutated p53. As suggested by the reviewer, in order to back up the AnnV/7AAD results we added WB for cleaved PARP, as described in response 3.	2	1	4) Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib ( when compared to the cells treated with TTFiealds and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 ( for both HCC cell lines). This might be helpful and make the in vitro data more relavant with the data shown in vivo.	1	2	cancers14122959_makarova	1
+We apologize for accidentally leaving out this figure, and have now added it.	2	1	1) Figure 4A is missing.	1	2	cancers14122959_makarova	1
+We thank the reviewer for this comment. In the in vitro experiments we used cell lines derived from humans. However, these cell lines cannot be implanted to rats, and so for the in vivo experiments we had to use a cell line from rats.	2	1	2) the different HCC cell lines were used for in vitro and in vivo experiments, therfore making difficult to compare these data.	1	2	cancers14122959_makarova	1
+We thank the reviewer for this important question and would be happy to clarify. TTFields are alternating electric fields in the range of 100 to 500 kHz, with maximal efficacy seen at a different frequency for different tumor types. Since TTFields cannot be applied at several different frequencies simultaneously, one specific frequency needs to be selected, and so the first step in examination of a new tumor type is a frequency scan. The effect of TTFields is not dichotomic, such that only one frequency is effective while the other are not at all. From the frequencies showing efficacy the purpose is to select the most effective one. It should be clarified that higher frequencies of the alternating electric fields do not mean higher energy, and hence there is no disadvantage in working with higher frequencies and no added toxicity or increased side effects for higher frequencies within the TTFields frequency range. Of note, TTFields at 200 kHz is already approved and has been applied to more than 18,000 patients with glioblastoma, with skin irritation being the main treatment related adverse event. TTFields at 150 kHz is approved for treatment of unresectable malignant pleural mesothelioma. To address these issues, we have rephrased the discussion: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” and “The lower TTFields intensity required for treatment of HepG2 relative to Huh-7D12 cells for achieving the same level of efficacy suggest higher sensitivity of the former to TTFields.” Point 2: What is the ‘n’ number of the samples and experiments in each group?	2	1	What is the rationale of using 150 kHz TTFields? TTFields at 100 kHz has also shown significant differences as cytotoxicity at p<0.01 and p<0.001 in HepG2 and Huh-7D12 cells which have around 60% and 50% of cell survival. Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).	1	2	cancers14122959_perova	1
+We thank the reviewer for pointing this was not clear. As mentioned in the statistical analysis sub-section within the methods part, all in vitro experiments were repeated at least 3 times, and so depicted We have now added this also to the legends of figures 1, 2, and 3. Per the in vivo study, as was mentioned in the methods (sub-section 2.8), 52 animals were included in the study. The specific numbers in each treatment group are now mentioned in the results (sub-section 3.4): “During the treatment period, average tumor volumes of control animals (n = 11) increased 5.9-fold (Figure 4b and 4c; and Figure S3 for tumor images). For animals treated with TTFields (n = 15) or sorafenib (n = 10), tumor growth was significantly lower, 3.3- and 2.3-fold, respectively. In the TTFields-sorafenib combination group (n = 16), a 1.6-fold increase in tumor volume within the treatment period was observed, a growth significantly lower than that for control or for each treatment alone.” It is also mentioned in the legend of figure 4: “Rats (n = 52) were inoculated orthotopically with rat HCC N1S1 cells, and treated with sham-vehicle (n = 11), TTFields alone (n = 15), sorafenib alone (n = 10), or TTFields + sorafenib (n = 16), according to the depicted timeline (a).” Point 3: LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group.	2	1	What is the ‘n’ number of the samples and experiments in each group? Also mention the ‘n’ number in all the experiments involved for invitro and invivo.	1	2	cancers14122959_perova	1
+First, we would like to emphasize that indications of autophagy were diverse and determined from increased cellular granularity, amplified lipidation of LC3 (from LC3-I to LC3-II) detected by Western blot, and elevated levels of LC3 foci observed by fluorescent staining. Nevertheless, to better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and additional time points. As per the reviewer’s request, Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but elevated only 2-fold in the combination group (Figure 4e).	2	1	LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group. How can the authors correlate autophagy with apoptosis? It is not so trustworthy that the only expression of LC3B indicate the treated condition have increased autophagy in the tumor tissue. Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.	1	2	cancers14122959_perova	1
+Per the reviewer’s request we show below images of the tumors within the liver, which we have also provided now in the supporting material of the paper. As may be seen, the liver itself is much larger than the tumor, and so we did not see value in measuring total liver weight, and hence cannot provide these values. While we tried to reduce variability between the animals by excluding those that at treatment initiation had tumor volumes not within the range 30–100 mm3, there was still divergence between the animals that resulted in the standard deviation bars depicted in the figure. It is however important to mention that there was a very good correlation between tumor volume (as measured by MRI) versus tumor weight at the end of the study, as may be seen in the graph below. Figure S3. Images of tumors within the livers of the rats treated with sham heat (control rats), TTFields, sorafenib, or TTFields plus sorafenib.	2	1	What was the total weight of livers in all groups? Please update the images of whole liver showing tumors on it, or at least pictures of tumors which were removed after sacrifice, if applicable.	1	2	cancers14122959_perova	1
+IHC tests the presence of a protein already expressed in the tissue, and 30 min are sufficient to allow recognition and binding of the antibody to the target protein. The secondary antibody is an HRP conjugated goat anti rabbit, that is a part of the Leica HRP-refine detection kit (Cat # DS9800). We have added the missing antibody information to section 2.10. We unfortunately do not have any tissue left to allow for performing WB analysis from the in vivo study, but we have conducted additional cell line studies and added WB analysis of cleaved PARP for the in vitro examinations and additional IHC examinations for autophagy and ER stress markers (as shown in response 3).	2	1	In the IHC experiment for PARP, the authors have incubated with primary antibody only for 30 min? Is that timing enough to get protein expression? What source of secondary antibody was used? In addition to IHC, I suggest performing western blot using PARP antibody where the full length and cleaved bands are observed in the same blot.	1	2	cancers14122959_perova	1
+We thank the reviewer for identifying this mistake, and have corrected this, changing the label of the LC3 data to e.	2	1	The figure number is mislabeled in Fig 4D-LC3 Immunofluorescence.	1	2	cancers14122959_perova	1
+Error bars are not missing for the control groups, rather they are null, as the values displayed in these figures are relative to control, and so per definition all control experiments have the exact same value of 100 or 1 (for percentage or fold change, respectively).	2	1	Why there is no error bar in the control group of all bar graphs? Please include error bars and re-calculate the statistical analysis for all the data wherever missing.	1	2	cancers14122959_perova	1
+We apologize for this mistake, and have now corrected the figure legend. The quantification of immunofluorescence of LC3 foci formation is c, and immunoblotting showing LC3-II to LC3-I ratio is d. Point 9: There is no data in Fig 4A.	2	1	Figure legends of Fig2D is missing. Fig C is repeated in the legend. Please label the figures appropriately. It is so frustrating to understand.	1	2	cancers14122959_perova	1
+We apologize for accidently omitting Fig 4A. This figure depicts the timeline of the in vivo experiment for easier understanding of the study designed. We have now corrected this.	2	1	There is no data in Fig 4A. Whole data is missing but have explained in the result section and in figure legends.??	1	2	cancers14122959_perova	1
+We understand from the reviewer’s comments that the sentence “The higher effects seen in the presence of CQ reveal that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation” is not clear enough. We have hence rephrased the sentence to be more accurate and better deliver the message: “CQ is an inhibitor of lysosome degradation, commonly used to decipher whether the elevation of LC3 is due to upregulation of the autophagy process or reduced autophagosome turnover [30]. The higher TTFields-induced elevation of LC3 seen in the presence of CQ suggest that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation.” Point 11: Typos: Line 111, ‘invitro’ spelling is not correct.	2	1	Line 337 seems over statement since the data are not shown in the manuscript.	1	2	cancers14122959_perova	1
+The “inovitro” in line 111 is not a typo, it is the name of the system used for applying TTFields in vitro. In line 342 we removed the duplicate “and”, and thank the reviewer for catching this typo.	2	1	Typos: Line 111, ‘invitro’ spelling is not correct. Double ‘and’ in Line 342. Need language and grammar check.	1	2	cancers14122959_perova	1
+For the in vitro experiments with TTFields (sub-section 2.3) we tried not to elaborate too much and referenced previous work describing all details. We understand that we have cut off too many details, and are happy to add them “HepG2 and Huh-7D12 cell suspensions (500 µl, 25 x 103 cells/plate) were placed as a drop in the center of 35-mm inovitro™ dishes composed of high dielectric constant ceramic (lead magnesium niobate–lead titanate [PMN-PT]), with two perpendicularly pairs of transducer arrays printed on their outer walls. Cells were incubated overnight at 37 °C to allow attachment to the dish, and then 2 ml of fresh media were added.” Regarding the number of rats included in the final analysis, we apologize this information was not clear. The 52 rats mentioned in the text were in fact the actual final number of animals in the analysis, since “all rats reached the required usage limit of ≥18 h/day”, as is now explicitly stated in the results sub-section 3.4.” Point 2: What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells.	2	1	In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells). A very important missing is not showing the actual number of rats included in the final analyses (the ones who successfully received therapy for more than 18 hours/day). This must be added.	1	2	cancers14122959_perova	1
+We thank the reviewer for raising this important issue. Using a xerograph model is indeed a good suggestion, however it would require working in immunosuppressed mice. It is currently not feasible for us to perform studies in which we apply TTFields to immunosuppressed animals, as these animals are of smaller weight thus the burden of wearing the electrodes may induce too much stress. Furthermore, these animals are more prone to contract infections during the process of electrodes placement and replacement on the animals. On the other hand, using the rat cells for in vitro experiments in the inovitro dishes turned out to be very technically challenging, as the cells were non-adherent, and so we could not pursue this important avenue. Nevertheless, we agree with the reviewer that there is a possibility that the optimal frequency may differ in the N1S1 relative to the human cell lines. Therefore, we have addressed this issue by including results of an additional frequency scan experiments done with N1S1 cells. We now mention this in the text, results sub-section 3.4: “The efficacy of combining TTFields with sorafenib relative to each modality alone was examined in the N1S1 HCC rat orthotopic model (timeline in Figure 4a). In vitro experiments confirmed that the 150 kHz TTFields frequency found optimal for treatment of the human cell lines was also optimal for treatment of the murine N1S1 cells used for the in vivo study (Figure S2).” We provide the relevant frequency scan figure in the supplementary material. Per the good question by the reviewer regarding the p53 status of the N1S1 cells we added: “It is also worth mentioning that the N1S1 murine cells used for this study, like the HepG2 cells, are p53 wild type.” Figure S2. TTFields frequency scan in rat N1S1 HCC cells. N1S1 cells were treated with TTFields (1.7 V/cm RMS) across a frequency range of 100–400 kHz, and cell count were determined following 24 hours of treatment. *p < 0.05 relative to control; one-way ANOVA. ANOVA = analysis of variance; HCC = hepatocellular carcinoma; RMS = root mean square; SEM = standard error of the mean; TTFields = Tumor Treating Fields.	2	1	What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells. While I do not know if the authors have the technology to perform TTFileds in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells. This must be done and included for a better understanding of TTFieldds activity from in vitro to in vivo data. What are the p53 status and the apoptosis signaling pathway function in N1S1 cells? Maybe the cytotoxicity data will reveal a different better frequency.	1	2	cancers14122959_perova	1
+We thank the reviewer for this comment, and apologize that the illustration of the timeline was accidentally missing from the submission (we have now added it). The limiting factor for study duration was the well being of the animals. The tumors in the control group were very large, causing stress and weight loss of the animals. The arrays placed on the animals, together with the individual housing needed to prevent wire entanglement, adds even more stress and increases animal weight loss. Overall, it was non-ethical to continue the study further. We have now added this explanation to the discussion section to clarify this limitation of the study: “In the HCC animal model, the acute effects of TTFields and sorafenib were examined. Due to the large tumors developed in the control group and the stress experienced by the animals as a result of the individual housing and motility limitations imposed by the sham and TTFields arrays, longer treatment durations were not feasible.” Point 4: There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing.	2	1	Another puzzling experiment is the schedule for the in vivo work. Since the authors missed to add Figure 4A for timeline, based on Methods section the rats were treated for 5 days with TTFields and or sorafenib and a day later the rays were sacrificed. While a short “acute” follow-up is welcome the most important experiment should allow the follow-up for much more days to indeed observe the effect of TTFields added to sorafenib. I could not find an explanation for not including a long term follow-up. In my opinion, this is a key therapeutically experiment which must be done and included in the study.	1	2	cancers14122959_perova	1
+It is important to understand that the TTFields frequency is not the treatment dose, and hence increased frequency does not mean increased efficacy. In the case of TTFields, intensity, duration, and usage are the factors that contribute to the “treatment dose”. Nevertheless, we have added to the discussion an explanation on this issue of optimal frequency: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” Regarding the option that the optimal frequency will differ between in vitro and in vivo setting, this is indeed a concern. However, as shown by the in vitro frequency scans, the effect of TTFields is not dichotomic, and effectiveness can be seen at more than one frequency. Therefore, the relatively low expected differences between animals treated with different TTFields frequencies together with the variability inherent to animal studies, makes it technically problematic to perform frequency scans in vivo. As of today, in vitro frequency scans are the common practice for determining the optimal TTFields frequency for delivery to animals and to humans. In fact, in vitro frequency scans were the basis for the clinical studies leading to FDA approval of TTFields at 200 kHz for the treatment of GBM and at 150 kHz for the treatment of MPM. We have added this issue to the discussion as a limitation of the study: “Since the optimal TTFields frequency has been shown to be dependent on the electrical properties of the cells and it is not clear how much effect the tumor microenvironment has on these properties, and because it is technically problematic to perform TTFields frequency scans in vivo, the frequency detected in the cell cultures was also employed for the animal studies.” Point 5: Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	2	1	There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing. Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is total different?	1	2	cancers14122959_perova	1
+When it comes to testing efficacy, we would like to use a treatment duration as long as possible to maximize the effect. However, this duration varies between in vitro and in vivo work. For cell cultures, the control samples continue to grow throughout the treatment duration, and so after 72 h the plates are very much confluent, and may not be left to continue and grow any further (without drastic changes in the environmental conditions, making them inadequate control cells). It is also noteworthy that such treatment periods for in vitro work are well accepted in the field of TTFields research. In vivo, prior studies also used similar time frames of 2 to 3 weeks from inoculation when working with the N1S1 model (Buijs et al., 2012 -; Ju et al., 2009; Thompson et al., 2012). Extending treatment duration was also limited by the physical status of the animals, as was explained in response 3.	2	1	Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	1	2	cancers14122959_perova	1
+Chloroquine was added to the cell cultures only at the final hours of the treatment for answering questions related to the mechanism of action, and not for boosting the efficacy of the other treatments. When using chloroquine in animal studies, it is for efficacy purposes, and so it is used throughout the treatment period. Since our animal experiment aimed to examine the efficacy of concomitant TTFields and sorafenib, with no additional agents, chloroquine was not employed.	2	1	Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.	1	2	cancers14122959_perova	1
+For the cytotoxicity assay we remove the supernatant, washed the cells, and then collect the adherent cells following trypsinization and visual inspection to verify all cells were removed. Indeed, as the reviewer mentioned correctly, we “counted the live cells and plotted the final data as percentage of untreated controls”. We agree that this is not cytotoxicity per se, and that is why we also perform 7-AAD/annexin-V staining of the cells. For this apoptosis assay we do collect the supernatant together with the adherent cells. Increased apoptosis and/or necrosis indicates that reduction in the cell number observed in the cell count emanates at least in part from cytotoxicity. We agree with the reviewer that we should not confuse the cell count measurements with the term “cytotoxicity” prior to showing the effect on apoptosis, and therefore we have changed this terminology throughout the paper.	2	1	The authors claim that cytotoxicity was measured “by cell counting using iCyt EC800 (Sony Biotechnology) 123 flow cytometer, and expressed as a percentage relative to the control.” Does this imply that they counted the live cells and plotted the final data as percentage of untreated controls (as figure 1 suggests). However, is this a real cytotoxicity or a cell growth inhibition? Did they measure the adherent cells after trypsinization. Were the cells from supernatant counted (where are probably the majority of dead cells)? There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.	1	2	cancers14122959_perova	1
+We thank the reviewer for these questions. Preliminary tests using CD31 staining revealed no differences between the groups regarding blood vessel density and therefore we did not pursue the research in that direction. We agree that measuring the effect of TTFields on the anti-angiogenic effect of sorafenib and on resistance to sorafenib are interesting, these topics were however not within the scope of the current study, and remain for future investigations. Regarding possible discrepancy between the volume fold change and tumor weight, please see below a graph showing tumor volume (as measured by MRI) versus tumor weight, both measured at the end of the study. The graph shows very good correlation between the two parameters, indicating the reliability of the measurements. If the reviewer feels there is discrepancy, it may be due to the volume shown as fold change relative to the initial tumor volume and not as the end value.	2	1	Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated? Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic? Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group. Did the authors check changes in blood vessels density. Were the mice perfused before collecting the tumors?	1	2	cancers14122959_perova	1
+We routinely examine our cells for Mycoplasma. Regarding authenticity, the cells were used shortly after purchase, and so there was no need to examine this.	2	1	Was Mycoplasma testing done routinely? Were the cells checked also for authenticity?	1	2	cancers14122959_perova	1
+We appreciate this question. Autophagy is a process that is elevated in order to cope with cellular stress, but as stress level elevate, autophagy can no longer provide the protection the cell needs, and the cell will undergo apoptosis. This kind of kinetics means that the levels of autophagy markers will depend on the time point the cells are examined. This may be appreciated from figure 2 panel c and d, with the different kinetics displayed by the two cell lines. While for Huh-7D12 autophagy levels increase from 24 to 48 hours of treatment, in the HepG2 cells autophagy levels at 48 hours are lower than at 24 hours, indicating these cells are already after the autophagy peak and on the way to apoptosis. Indeed, figure 1d shows higher levels of apoptosis for HepG2 cells. To clarify we have added a few sentences to the text. In results sub-section 3.2: “However, autophagy kinetics seems to be faster in the HepG2 cells, in which LC3 markers are lower at 48 versus 24 hours, whereas elevation is seen from 24 to 48 hours for the Huh-7D12 cells.” In the discussion “While autophagy serves as a survival strategy of cells, when stress levels continue raising it may be over-activated and mediate cell death [9]. The faster autophagy kinetics seen for the HepG2 relative to Huh-7D12 cells following application of TTFields is in agreement with the higher apoptosis levels displayed by this cell line, and may serve as an additional rational for the higher efficacy of TTFields against it. Examination of the reasons for faster autophagy in HepG2 relative to Huh-7D12 cells is out of the scope of this work.” For more clarity we have also added a more in-depth kinetic study, including additional relevant markers and additional time points, examinations that were performed for the combined treatment as compared to TTFields and sorafenib alone. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case” Figure 3. In the animal study we may only measure one time point. The lower autophagy seen at this time for the combined treatment together with the higher apoptosis indicate that we are further along the kinetic timeline of autophagy relative to the monotherapies, suggesting higher stress in animals receiving the combined treatment. We have now added IHC examination of beclin-1, an additional autophagy marker, and of GRP78, a marker for ER stress, as described in results sub-section 3.4.	2	1	Is there any explanation why the combination of TTFields and sorafenib did not induce a significant level of autophagy as compared to untreated animals which invalid the initial hypothesis that “concomitant application of sorafenib and TTFields may increase stress levels enough to tilt autophagy towards the cell death pathway”.	1	2	cancers14122959_perova	1
+The sentence referred to was meant to describe only the in vivo outcomes. We have rephrased it to be more accurate and clear: “While each treatment alone elevated levels of autophagy relative to control, TTFields concomitant with sorafenib induced a significant increase versus control in tumor ER stress and apoptosis levels, demonstrating increased stress under the multimodal treatment.” Point 12: Finally, adding to all the above questions, I found a very weak Discussion section which must be extended.	2	1	The statement “TTFields concomitant with sorafenib induced a significant increase in apoptosis’ in the abstract section is overstated. When compared with sorafenib alone there is practically no difference. Moreover, TTFields failed to increases apoptosis when added to sorafenib and compared to sorafenib alone in one out of two human cells line investigated	1	2	cancers14122959_perova	1
+We thanks the reviewer for this comment. We have now elaborated on many issues throughout the discussion, as was described thorough this response letter. Regarding the conclusion, we have rephrased it for better accuracy: “TTFields were identified to be most efficient for treatment of HCC cells at 150 kHz, and this frequency further demonstrated in vivo efficacy.” Why only one frequency was used in vivo, and the difference between frequency and dose, were explained in response 4.	2	1	Finally, adding to all the above questions, I found a very weak Discussion section which must be extended. Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”. This is not a correct statement. While for in vitro data, the authors have data, for in vivo they used only one frequency of 150 Hz. At least one different dose should have been studied for comparison since this is a completely different tumor environment than the in vitro one.	1	2	cancers14122959_perova	1
+We thank the reviewer for this question. Indeed, the sorafenib dose used in this study proved to be slightly more efficacious than TTFields in controlling tumor fold increase. Nevertheless, these differences between the monotherapies, did not reach statistical significance. In accordance, there was no statistical difference between the monotherapies in the expression levels of the LC3 marker and the levels of cleaved PARP. In order to better understand the mechanism of action, we have added experiments to better characterize the autophagy-apoptosis interplay for treatment with concomitant TTFields and sorafenib, which are described in response 3. Of note, the clinical development of TTFields does not aim to replace sorafenib with TTFields, but rather to add TTFields on top of sorafenib and therefore this work focused mainly on the potential added value in combining these 2 modalities.	2	1	1) The authors demonstrate the efficacy of TTFields in vivo even when used as monotherapy. As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib. However, no differences in expression of LC3 marker were observed between these groups (treated with TTFields or with sorafenib)(as shown in Figure 4D). Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F. What is the mechanism illustrating higher efficiency of sorafenib against HCC?	1	2	cancers14122959_perova	1
+We thank the reviewer for pointing out this issue. Quantification of the IHC images was done automatically. The whole slide was scanned, and the CaseViewer software was used to exclude non-tumor areas. The signals of the stained protein and the nuclei were resolved by color deconvolution and quantified separately using the FIJI software (ImageJ) software. Average signal per cell or percent of positive cells was calculated. As the reviewer pointed out, the high magnification images we chose to show do not correctly reflect the quantification performed by the software, and we have now replaced them with better representative fields of the slides.	2	1	2) Despite the expression of cleaved PARP was very low in the tumors treated with TTFields or sorafenib alone ( as shown in IHC-images in Figure 4F), the authors declare about ~ 20% of positive cells, as show in the graphs below IHC-staining. Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.	1	2	cancers14122959_perova	1
+values are mean (N ≥ 3) ± SEM. We sincerely appreciate this well-taken comment. these examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3. HepG2 cells were treated for 6, 24, or 48 hours with 150 kHz TTFields, 3 µM sorafenib, or the two treatments combined, followed by Western blot examination of the autophagy markers beclin-1 and LC3 (d), the ER stress marker GRP78 (e), and the apoptosis marker cleaved PARP (f). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA. In the animal study we have now added IHC examination of beclin-1 and of GRP78, a marker for ER stress, as described in results sub-section 3.4. Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively. GRP78 levels in the groups treated with TTFields or sorafenib alone remained unchanged from the control, but were elevated 2-fold in the TTFields plus sorafenib group (Figure 4f). Additionally, the percentage of cells positive for cleaved PARP was significantly higher relative to control only in the combination group (Figure 4g).” and also in the discussion: “The lower autophagy accompanied by the higher ER stress and apoptosis displayed in the conjunction group relative to the monotherapies groups following 6 days of treatment suggest that these animals were pushed further along the autophagy-apoptosis kinetic timeline due to the higher levels of stress experienced by these animals, in accordance with the results described for the cell cultures.” Figure 4. tumor slices were subjected to immunohistochemical analysis for beclin-1 and LC3 (e), GRP78 (f), and cleaved PARP (g). Values are mean ± SD. *p < 0.05, **p < 0.01, and ****p < 0.0001 relative to control for labels above bars, or between indicated groups; Student’s T-test. Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but only 2-fold in the combination group (Figure 4e). To better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and at additional time points.	2	1	3) It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenin induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.	1	2	cancers14122959_perova	1
+Both cell lines experience elevation of apoptosis following application of sorafenib in a dose dependent manner, as evident from the AnnV/7AAD results. However, while TTFields greatly elevate apoptosis in HepG2 cells, they have a low effect on apoptosis levels in the Huh-7D12 cells, seen both in Figure 1d and in Figure 3c. As was explained in the discussion, this difference between the cell lines may be attributed to the different p53 status, wild type in HepG2 and mutated in Huh-7D12, as there are previous indications of lower TTFields-induced apoptosis in cell lines with mutated p53. As suggested by the reviewer, in order to back up the AnnV/7AAD results we added WB for cleaved PARP, as described in response 3.	2	1	4) Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib ( when compared to the cells treated with TTFiealds and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 ( for both HCC cell lines). This might be helpful and make the in vitro data more relavant with the data shown in vivo.	1	2	cancers14122959_perova	1
+We apologize for accidentally leaving out this figure, and have now added it.	2	1	1) Figure 4A is missing.	1	2	cancers14122959_perova	1
+We thank the reviewer for this comment. In the in vitro experiments we used cell lines derived from humans. However, these cell lines cannot be implanted to rats, and so for the in vivo experiments we had to use a cell line from rats.	2	1	2) the different HCC cell lines were used for in vitro and in vivo experiments, therfore making difficult to compare these data.	1	2	cancers14122959_perova	1
+We have introduced new several sentences in this paragraph of Materials and Methods, which define these ad-hoc rules. Lines 154-155: “mgR51C read-outs have been introduced into the classification system as PVS1_O or BP7_O codes of variable evidence strength depending on the splicing outcome [P, Sup-porting (±1 point); M, Moderate (±2); Strong (±4); Very Strong (±8)].” Lines 163-179: “we have developed some ad-hoc rules that take into consideration the coding potential of each individual transcript and its relative contribution to the overall expression to reach the appropriate PVS1_O or BP/_O evidence strength. In brief, for each complex read-out we have applied the following algorithm: (i) De-convolute mgR51C read-outs in-to individual transcripts; (ii) apply ACMG/AMP evidences to each individual transcript; (iii) produce an overall PVS1_O (or BP7_O) code strength based on the relative contribu-tion of individual transcripts/evidences to the overall expression. Thus, if pathogenic supporting transcripts contribute ≥90% to the overall expression, PVS1_O_ code is applied (if different transcripts support different pathogenic evidence strengths, the lowest strength contributing >10% to the overall expression is selected for overall evidence strength). Similarly, BP7_O_ code is applied if benign supporting transcripts contribute ≥90% to the overall expression (if different transcripts support different pathogenic evi-dence strengths, the lowest strength contributing >10% to the overall expression is selected for overall evidence strength). If neither pathogenic nor benign supporting transcripts contribute ≥90% to the overall expression, the splicing assay is considered not providing any evidence in favor, or against, pathogenicity. Recently, we have used a similar ap-proach to deal with complex PALB2/ATM minigene read-outs [20,30].” 2 -	2	1	Line 156 – the authors refer to “ad-hoc rules” that they have developed for consideration of the different coding transcripts associated with the same spliceogenic variant in variant interpretation and classification. Although a reference is provided so that the reader can look up what these ad-hoc rules are, it would also be helpful to briefly describe these in the current manuscript.	1	2	cancers14122960_makarova	1
+The five RAD51 paralogs are known to be required for homologous recombination and maintenance of genomic stability. Indeed, RAD51C interacts with RAD51B, RAD51D, XRCC2 and XRCC3 in two different complexes that play a role in homologous recombination. Miller et al studied the interaction between RAD51B and D (and also XRCC3) with deletion mutants. These authors found that Rad51C1-285 (includes β-strands 1-5) or Rad51C285-376 (includes β-strands 6-9) did not bind RAD51B. So, a complete beta-sheet is important in maintaining the overall fold of the protein. Moreover, the missense variant p.Arg312Trp (ß-strand 6) has been shown to impair RAD51C function (Gayarre et al 2017). Both studies indicate that this protein region is essential for RAD51C function so that transcripts lacking any of the β-strands, such as ▼(E6q4)-a, ▼(E6q4)-b, Δ(E7), Δ(E8) or the in-frame isoform Δ(E5), is probably deleterious. - We have modified this part, adding several sentences to clarify it. Lines 270-277: “ The integrity of the β-sheet is important in maintaining the overall fold of the RAD51C protein and the interaction with RAD51B, so that alterations of any ß-strand of RAD51C should be considered deleterious [33]. Further, structural features (the order of the ß--strands in space is not the same as their order in sequence) predict that proteins lacking any single b-strand would fail to form the ß--sheet resulting in a collapse of the protein core and misfolding of the protein [33]. Moreover, the missense variant p.Arg312Trp (ß-strand 6) has been shown to impair RAD51C function [34]. Altogether these data,…” 3-	2	1	Lines 250-251 and 263-265– In these lines the authors discuss transcripts in which the encoded proteins lack some beta strands. The lack of these protein structures are used as evidence to support pathogenicity. It is not clear from the text how lack of these beta strands is predicted to impact protein function. Is there evidence from another source that these beta strands are critical to protein function and that their loss is deleterious (rather than resulting in normal or slightly reduced protein activity)?	1	2	cancers14122960_makarova	1
+We have added all the transcript names and their contribution in Table 2 (Column PVS1_O/BP7_O mgR51C_ex2-8).	2	1	Lines 259-274 – This paragraph refers to evidence used in the variant interpretation of three aberrant transcripts that kept the open reading-frame (Δ(E2p3), Δ(E5) and ▼(E8p3)). However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names. As such, in order to correlate the discussion in this paragraph with the information in table 2, the reader also needs to cross reference Table 1 or 3. Incorporation of the transcript isoform names in table 2 would assist the reader in correlating this discussion of transcript isoforms with the corresponding evidence used to classify each of the variants.	1	2	cancers14122960_makarova	1
+We have included cross-references to Table 1 to facilitate understanding of the manuscript.	2	1	Lines 259-274 – This paragraph refers to evidence used in the variant interpretation of three aberrant transcripts that kept the open reading-frame (Δ(E2p3), Δ(E5) and ▼(E8p3)). However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names. As such, in order to correlate the discussion in this paragraph with the information in table 2, the reader also needs to cross reference Table 1 or 3. Incorporation of the transcript isoform names in table 2 would assist the reader in correlating this discussion of transcript isoforms with the corresponding evidence used to classify each of the variants.	1	2	cancers14122960_makarova	1
+Very important comment. As the reviewer indicates, it would be essential to define the threshold of RAD51C expression from which it keeps its tumor suppressor activity. Unfortunately, it is not known by now but this finding would provide critical information to determine the pathogenicity of leaky spliceogenic variants.	2	1	Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively. The discussion of these variants in this section (and in other sections) does not address the significance of canonical transcripts in these cases. Leaky splice variants have been reported in various genes and sometimes can be associated with milder phenotypes (or no phenotypes), presumably because the canonical isoforms contribute to a “phenotypic rescue”. Is it known whether there is a threshold of RAD51C deficiency that is tolerated before associated cancer risks become increased?	1	2	cancers14122960_makarova	1
+So, we have modified this paragraph to introduce this information: “Only two variants (c.404+3A>G and c.705+3A>G) displayed 26% and 21% of the mgFL-transcript, respectively. Unfortunately, it is not known the minimal amount of RAD51C expression to confer tumor suppressor haplosufficiency so, these splicing assays were not considered informative (PVS1_O_N/A).” Note that leaky variants generate complex minigene read-outs (two or more transcripts), and are therefore classified accordingly (see methods).	2	1	Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively. The discussion of these variants in this section (and in other sections) does not address the significance of canonical transcripts in these cases. Leaky splice variants have been reported in various genes and sometimes can be associated with milder phenotypes (or no phenotypes), presumably because the canonical isoforms contribute to a “phenotypic rescue”. Is it known whether there is a threshold of RAD51C deficiency that is tolerated before associated cancer risks become increased?	1	2	cancers14122960_makarova	1
+Acknowledge this suggestion. We have added two references of BRCA1 and BRCA2 studies.	2	1	in the Introduction in the sentence 61-69 the authors do not mention anything about BRCA1 (they mention MLH1 though); I would expect that as BRCA1 splice variants are deeply studied.	1	2	cancers14122960_makarova	1
+In this section we have tried to describe the classification approach and the rules we have followed to classify the variants. Certainly, we agree with the referee as sometimes the method is mixed with some results. So, we have moved some sentences of the last paragraph of Materials and Methods to Results. “The PM3 evidence (in trans with a pathogenic variant in a recessive disorder) did not con-tribute to the final classification. Not surprisingly (FANCO is an extremely rare FA complementation group) [37], none of the tested variants has been identified in Fanconi Anemia patients (ClinVar and Global Variome shared LOVD databases and literature search-es). Similarly, the BS2 evidence (in trans with a pathogenic variant in a healthy individual) does not contribute to the final classification of our tested variants. Finally, we have considered that some pathogenic (PS2, PM1, PM6, PP2, PP4) and benign (BP1, BP3, BP5) codes are not applicable to the classification of RAD51C variants.” 3-	2	1	In the Materials and Methods, the section 2.6 is not actually a section that described some methodology. It appears to be more appropriate as results and maybe some information could be in the introduction.	1	2	cancers14122960_makarova	1
+We have included explanations of these acronyms in text and Table 1 and have modified the following sentence: “Of the 19 characterized transcripts, 14 introduced premature termination codons (PTC; PTC transcripts), and of these, 10 are predicted to be degraded by the Nonsense-Mediated Decay pathway (NMD; PTC-NMD transcripts) that is considered convincing evidence of deleteriousness (Supplementary Table S3).	2	1	The acronyms need to be explained. Not all readers are familiar with PTC (premature termination codon) and NMD (nonsense mediated decay); as some variants are found PTC-NMD, this needs to be explained better. I would also stress a little more that FL is almost undetectable.	1	2	cancers14122960_makarova	1
+We have modified the sentence of the Fl-transcript: “All variants altered splicing, 18 of which produced no traces of the mgFL-transcript or almost undetectable levels (<2.4%, c.904G>A)…”	2	1	The acronyms need to be explained. Not all readers are familiar with PTC (premature termination codon) and NMD (nonsense mediated decay); as some variants are found PTC-NMD, this needs to be explained better. I would also stress a little more that FL is almost undetectable.	1	2	cancers14122960_makarova	1
+Acknowledge this comment. Splicing variants are called solely based on the size difference. It is good to look into Sanger sequence of splice variants after gel elution of the band and sequencing. We have sequenced the RT-PCR products of all variant assays (indicated in Materials and Methods, section 2.5. Minigene Splicing Assays). In fact, all the *.ab1 sequence and*.fsa fragment analysis files of RT-PCR products will be freely available at http://hdl.handle.net/10261/270934; https://doi.org/10.20350/digitalCSIC/14662 upon manuscript acceptance (links indicated in the manuscript section “Data Availability Statement”). Unfortunately, Sanger sequencing only allowed us to characterize the main transcripts, while the minor ones (<10% of the overall expression) are really difficult to characterize since gel band extraction does not work properly with these small amounts, or other methods (e.g. subcloning of RT-PCR products into a PCR-vector, or RNAseq of minigene outcomes) are laborious and not cost-effective. Anyway, we have also been using Fluorescent fragment analysis for transcript characterization in our previous studies (since Acedo et al, 2012). We have shown that this technique is highly sensitive, accurate and shows high resolution. For example, in Figure 1c, transcripts with minimal size differences (1-3 nt) are well-discriminated. So, for minor-rare transcripts is a good option (not perfect, we agree with the reviewer) to annotate them. - To clarify it, we have modified this part: Lines 132-134: “RT-PCR products were sequenced by Macrogen (Madrid, Spain), which allowed the characterization of the main variant-induced transcripts. Minor transcripts were annotated according to fluorescent fragment electrophoresis size data (see below).”	2	1	Splicing variants are called solely based on the size difference. It is good to look into Sanger sequence of splice variants after gel elution of the band and sequencing.	1	2	cancers14122960_makarova	1
+We would like to thank reviewer #1 for his/her time and effort to review our manuscript. Please find our point-by-point responses below.	2	1	This manuscript adds evidence that NR2F1 is a dormancy marker. In addition, it shows that NR2F1 is primarily expressed in CAFs, particularly in inflammatory CAFs.	1	2	cancers14122962_makarova	1
+We totally agree with the reviewer that it is puzzling how NR2F1 expressed in CAFs of the primary breast cancer contribute to the dormancy of DTCs, thus the addition of a discussion on this point in more detail will strengthen this manuscript. Based on our results, NR2F1 expression in primary bulk tumor is associated with several pathways related to dormancy, and NR2F1 is most predominantly expressed in CAFs in the tumor microenvironment. However, we did not prove the underlying mechanism through which CAF-expressed NR2F1 regulates dormancy, and we do not intend to claim a causal relationship. Single-cell sequence data of metastatic tumor cohorts will allow us to investigate whether the expression of NR2F1 in CAFs in the metastatic TME is related to the dormancy of DTCs. We added the following sentences to the discussion section.	2	1	Given that dormant disseminated tumor cells (DTCs) highly express NR2F1 (Fluegel et al. Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs. It would be nice, if the authors would discuss this important point in more detail.	1	2	cancers14122962_makarova	1
+We agree with the reviewer that it would be interesting to see how NR2F1 levels would change with endocrine therapy. However, we do not have access to cohorts that include tumor samples before and after neoadjuvant endocrine therapy at this point. What we do have access to regarding endocrine therapy is a cohort comparing responders and non- responders to neoadjuvant endocrine therapy (GSE145325). We found that NR2F1 expression between responders and non-responders to endocrine therapy was not different. We added this to the results section as follows.	2	1	The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels. It would be interesting to see, how NR2F1 levels would change with endocrine treatment and/or endocrine resistance.	1	2	cancers14122962_makarova	1
+We agree with the reviewer that breast cancer is a heterogenous disease and we should show subtype-specific data. We analyzed survival outcomes and the cell fraction in TME by each immunohistological subtype. We did not observe any validated difference in survival outcomes. Cell fractionation of immune cells and stromal cells showed almost similar trends for the scores such as intratumor heterogeneity, HRD, mutation rate, and neoantigens across all subtypes. The results for each immunohistological subtype of single-cell Cohort 2 are shown in Supplementary Figure 7, and each subtype showed the same trend. We have revised the results section and added supplementary data as follows.	2	1	Breast cancer is a heterogenous disease. Different subtypes behave differently in many aspects. It would be great to see some subtype-specific data.	1	2	cancers14122962_makarova	1
+We thank the reviewer for reading our manuscript closely and pointing out our oversight. As you indicated, we did not find any significance difference in 2D in Figure 1A and have corrected the results section as follows.	2	1	The authors state “We demonstrated that the expression of NR2F1, RARB, and TGFB1 genes are higher in previously established dormant cells (D2OR murine breast cancer cells [44] compared to the proliferative cells (D2A1 cells) in both 2D and 3D cultures (Figure 1A, all p < 0.02).” This is not true for NR2F1 in 2D. Please correct.	1	2	cancers14122962_makarova	1
+We are grateful to the reviewer for her/his time and effort in reviewing our paper, as well as for pointing out issues to improve the paper.	2	1	The authors present interesting findings that a tumor dormancy marker, NR2F1, is predominantly expressed in the inflammatory CAFs, and high expression of NR2F1 is associated with suppressed immune response and increased density of stromal cells. However, this reviewer has a few concerns that need to be addressed before accepting this article for publication.	1	2	cancers14122962_makarova	1
+We completely agree with the reviewer that it will be informative to demonstrate the NR2F1 expression in both primary and metastatic breast cancer and the possible correlation of NR2F1 expression with metastasis. In Figure 3C, we present NR2F1 expression in primary breast cancer with and without distant metastases. NR2F1 expression was not increased in the group with later recurrence in four cohorts in this analysis. We also present NR2F1 expression between primary and metastatic breast cancer in Figure 3E with no significant difference. We did not find a clear association of NR2F1 with distant metastasis in this study. On the other hand, NR2F1 expression was higher in primary breast tumors with lymph node metastasis in all four cohorts, as shown in Figure 3B, suggesting an association between NR2F1 and lymph node metastasis. Given these results, we revised the results section as follows.	2	1	This reviewer noticed that all the analysis was performed on the public data sets of bulk RNA-seq or single cell sequencing of primary breast tumors. Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?	1	2	cancers14122962_makarova	1
+We completely agree with the reviewer that NR2F1 expression in cancer cells and CAFs should be investigated not only in primary but also in metastatic breast cancer using single-cell sequence in order to prove that NR2F1 expression in CAFs affects late recurrence. The main finding of this study is that NR2F1 is predominantly expressed in CAFs rather than in all other cell types in the TME, and we do not intend to claim that CAF-expressed NR2F1 regulates dormancy. Further, we do not have access to single-cell sequence cohorts of metastatic breast cancer tumors, but it is of our interest, and this will be our future direction. In response to the reviewer, we added the following sentences in the discussion section.	2	1	Moreover, the authors need to provide some, at least minimum, evidence or clues that CAF-expressed NR2F1 is responsible for tumor dormancy regulation.	1	2	cancers14122962_makarova	1
+We thank the reviewer for closely reading our manuscript and pointing out our oversight. We have corrected the results section as answered in Responce1.	2	1	Line 170, “Figure 1E” should be “Figure 1D”.	1	2	cancers14122962_makarova	1
+We totally agree with the reviewer that it is puzzling how NR2F1 expressed in CAFs of the primary breast cancer contribute to the dormancy of DTCs, thus the addition of a discussion on this point in more detail will strengthen this manuscript. Based on our results, NR2F1 expression in primary bulk tumor is associated with several pathways related to dormancy, and NR2F1 is most predominantly expressed in CAFs in the tumor microenvironment. However, we did not prove the underlying mechanism through which CAF-expressed NR2F1 regulates dormancy, and we do not intend to claim a causal relationship. Single-cell sequence data of metastatic tumor cohorts will allow us to investigate whether the expression of NR2F1 in CAFs in the metastatic TME is related to the dormancy of DTCs. We added the following sentences to the discussion section.	2	1	Given that dormant disseminated tumor cells (DTCs) highly express NR2F1 (Fluegel et al. Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs. It would be nice, if the authors would discuss this important point in more detail.	1	2	cancers14122962_perova	1
+We agree with the reviewer that it would be interesting to see how NR2F1 levels would change with endocrine therapy. However, we do not have access to cohorts that include tumor samples before and after neoadjuvant endocrine therapy at this point. What we do have access to regarding endocrine therapy is a cohort comparing responders and non- responders to neoadjuvant endocrine therapy (GSE145325). We found that NR2F1 expression between responders and non-responders to endocrine therapy was not different. We added this to the results section as follows.	2	1	The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels. It would be interesting to see, how NR2F1 levels would change with endocrine treatment and/or endocrine resistance.	1	2	cancers14122962_perova	1
+We agree with the reviewer that breast cancer is a heterogenous disease and we should show subtype-specific data. We analyzed survival outcomes and the cell fraction in TME by each immunohistological subtype. We did not observe any validated difference in survival outcomes. Cell fractionation of immune cells and stromal cells showed almost similar trends for the scores such as intratumor heterogeneity, HRD, mutation rate, and neoantigens across all subtypes. The results for each immunohistological subtype of single-cell Cohort 2 are shown in Supplementary Figure 7, and each subtype showed the same trend. We have revised the results section and added supplementary data as follows.	2	1	Breast cancer is a heterogenous disease. Different subtypes behave differently in many aspects. It would be great to see some subtype-specific data.	1	2	cancers14122962_perova	1
+We thank the reviewer for reading our manuscript closely and pointing out our oversight. As you indicated, we did not find any significance difference in 2D in Figure 1A and have corrected the results section as follows.	2	1	The authors state “We demonstrated that the expression of NR2F1, RARB, and TGFB1 genes are higher in previously established dormant cells (D2OR murine breast cancer cells [44] compared to the proliferative cells (D2A1 cells) in both 2D and 3D cultures (Figure 1A, all p < 0.02).” This is not true for NR2F1 in 2D. Please correct.	1	2	cancers14122962_perova	1
+We completely agree with the reviewer that it will be informative to demonstrate the NR2F1 expression in both primary and metastatic breast cancer and the possible correlation of NR2F1 expression with metastasis. In Figure 3C, we present NR2F1 expression in primary breast cancer with and without distant metastases. NR2F1 expression was not increased in the group with later recurrence in four cohorts in this analysis. We also present NR2F1 expression between primary and metastatic breast cancer in Figure 3E with no significant difference. We did not find a clear association of NR2F1 with distant metastasis in this study. On the other hand, NR2F1 expression was higher in primary breast tumors with lymph node metastasis in all four cohorts, as shown in Figure 3B, suggesting an association between NR2F1 and lymph node metastasis. Given these results, we revised the results section as follows.	2	1	This reviewer noticed that all the analysis was performed on the public data sets of bulk RNA-seq or single cell sequencing of primary breast tumors. Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?	1	2	cancers14122962_perova	1
+We completely agree with the reviewer that NR2F1 expression in cancer cells and CAFs should be investigated not only in primary but also in metastatic breast cancer using single-cell sequence in order to prove that NR2F1 expression in CAFs affects late recurrence. The main finding of this study is that NR2F1 is predominantly expressed in CAFs rather than in all other cell types in the TME, and we do not intend to claim that CAF-expressed NR2F1 regulates dormancy. Further, we do not have access to single-cell sequence cohorts of metastatic breast cancer tumors, but it is of our interest, and this will be our future direction. In response to the reviewer, we added the following sentences in the discussion section.	2	1	Moreover, the authors need to provide some, at least minimum, evidence or clues that CAF-expressed NR2F1 is responsible for tumor dormancy regulation.	1	2	cancers14122962_perova	1
+We thank the reviewer for closely reading our manuscript and pointing out our oversight. We have corrected the results section as answered in Responce1.	2	1	Line 170, “Figure 1E” should be “Figure 1D”.	1	2	cancers14122962_perova	1
+According to the reviewer suggestion, FT-IR peaks shift related to chromium ion adsorption are discussed in the mechanism section and is incorporated in the revised manuscript.	2	1	Some FT-IR peaks shift with adsorption of Cr, and some not. Please add the discussion which peak shift is related to Cr, the authors can combine these with the discussion of the mechanism.	1	2	catal12030290_makarova	1
+EDX analysis (In Figure 3) – is a conditional type of analysis of the chemical composition of the surface. In EDX, often, spectra with different atomic abundances of elements can be obtained even from the same sample.	2	1	In Figure 3, what is the origin of the increase concentration of Al and S?	1	2	catal12030290_makarova	1
+The sulphuric acid disintegrated the leaves. Most of the substances in the leaves are reduced to carbon after two hours. Charring takes place by adding sulphuric acid and by the action of heat, charring removes hydrogen and oxygen from the solid, so that the remaining char is composed primarily of carbon. It also helps to remove the moist content in the leaves. The pollution problem may not be takes place.	2	1	In the experiment, H2SO4 is added in the first step, what is the purpose? Will this bring pollution problem?	1	2	catal12030290_makarova	1
+Chromium is a potentially toxic metal occurring in water and groundwater as a result of natural and anthropogenic sources. The prepared adsorbent is well suitable in the real condition. Since the prepared adsorbent (Al-GNSC) is successfully reduces the chromium (VI) ion form the real groundwater samples.	2	1	How about the Cr concentration in the polluted ground water? Is the products suitable for this real condition?	1	2	catal12030290_makarova	1
+Based on the reviewer suggestion, regeneration studies were performed briefly. Four different desorption agents such as tap water, 0.1M HCl, 0.1M H2SO4 and 0.1M NaOH were utilized to remove the adsorbed chromium ions from the Al-GNSC adsorbent. From this various desorption agents it was identified that 0.1M NaOH was more effective. Hence, the reuse of Al-GNSC from Cr(VI)-loaded material was studied for sorption and desorption cycles using sodium hydroxide as a regenerated agent. Relevant references are quoted in the revised manuscript. Reference: Sujitha Ravulapalli and Ravindhranath Kunta reported that sodium hydroxide was used as a regenerating agent for the sorption and desorption of Cr(VI) by activated carbon derived from Lantana camara plant. [Enhanced removal of chromium (VI) from wastewater using active carbon derived from Lantana camara plant as adsorbent. Water Sci Technol (2018) 78 (6): 1377–1389. https://doi.org/10.2166/wst.2018.413] Reference: M.A. Tandal and B.N.OZA. reported that Sodium hydroxide as a regenerating agent for the sorption and desorption of Cr(VI) by Granular activated carbon. [Adsorption and regeneration studies for the removal of Chromium (VI) from the waste water of electroplating industry using Granular activated carbon. Asian Journal of chemistry. Vol. 17, No.4 (2005), 2524-2530] Page 2 of 2 Comment 2:	2	1	The issue of regeneration has not been sufficiently worked out, but the authors themselves write about this, and it is not clear why NaOH was used for this.	1	2	catal12030290_makarova	1
+We acknowledge the reviewer’s opinion. For the preparation of 1kg of Al-GNSC adsorbent, approximately 150 to 200 mL of hydrochloric acid was added in order to blend the aluminum in to the ground nut shell carbon. Thank you for your valuable comment.	2	1	It was also not clear to me how much hydrochloric acid was eventually added during the preparation of the adsorbent.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, the expression of separation factor is mentioned in the revised manuscript. 𝑅𝐿= 1 1+𝑏𝐶𝑖, where “Ci” is the initial concentration of Cr(VI) and “b” is the Langmuir constant.	2	1	The authors did not indicate how the “separation factor ‘RL’ is calculated and did not give a link to the equation for its calculation, and if the readers are not quite in the subject, then what kind of factor they do not understand.	1	2	catal12030290_makarova	1
+However, we regret that we were not able to investigate the BET analysis due to pandemic situation, which could definitely give us additional information.	2	1	One could also estimate the specific surface area and porosity. So, the paper can be published after revision.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, the complete terms of all abbreviations are mentioned before the first use in the revised manuscript.	2	1	The authors should write the complete terms of all abbreviations (including the instruments) before the first use in the abstract and main manuscript i.e. FT-IR and SEM in abstract section et al.	1	2	catal12030290_makarova	1
+As suggested, the novelty of the research work is explained in the introduction part of the revised manuscript, and the Page 2 of 4 obtained results are compared with the recent literature.	2	1	The authors should clearly explain the innovation and importance of their work on the introduction of the manuscript. They should justify the value of the work and compare their work with previously similar published papers.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, the SEM images with same scale are provided in the revised manuscript.	2	1	Fig. 1 - for a more effective visual comparison, authors recommended to provide SEM images of the same scale. In such form is rather difficult to make adequate comparison.	1	2	catal12030290_makarova	1
+We hope the reviewer understand the experimental deficiencies at the stage of the present experiments. We deeply appreciate the comment raised by the reviewer. Thank you very much. However, we regret that we were not able to investigate the XPS analysis due to pandemic situation, which could definitely give us additional information about the elemental confirmation.	2	1	First of all, authors have to attach a EDX mapping images before/after sorption of chromium ions. And the authors are strongly recommended to add XPS spectra to this section of revised manuscript. The XPS method is much more sensitive and more accurately determines changes in the chemical composition of samples.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, XRD study has been performed and incorporated in the revised manuscript.	2	1	Why authors did not use XRD technique for sample characterization?	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, kinetic study has been performed and incorporated in the revised manuscript.	2	1	The adsorption capacities of Al-GNSC adsorbents at different contact times have been provided. Which kinetics are right? Please add missing information about appropriate kinetic model in revised manuscript.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion, the conditions for testing adsorbents of Cr(VI) is incorporated in the Table and is mentioned in the revised manuscript.	2	1	It is rather difficult to make an adequate comparison of certain properties (catalysts or sorbents) with the already available results, since the concentration of the pollutant and the mass of the loaded sorbent vary in each experiment. Therefore, the authors are recommended to add the missing information (i.e. conditions for testing sorbents of Cr(VI) ions) to Table 3.	1	2	catal12030290_makarova	1
+According to the reviewer suggestion the adsorption capacity of groundnut shell activated carbon (Qe= 7.4 mg/g) is mentioned in the revised manuscript.	2	1	In order to confirm proposed mechanism of Cr(VI) adsorption (illustrated on the fig 6) Authors should provide data on adsorption capacity of pristine groundnut shell activated carbon (not modified with Al).	1	2	catal12030290_makarova	1
+Thank you for your valuable suggestion. According to the reviewer suggestion the conclusion section is elaborated with specific conclusions in the revised manuscript.	2	1	The conclusion section should be elaborated and improved. The author should bring specific conclusions in accordance with obtained results.	1	2	catal12030290_makarova	1
+We acknowledge the reviewer’s opinion. As suggested by the reviewer, we checked the manuscript carefully and enlisted a professional English language service to eliminate the spelling mistakes and grammatical errors. Thank you for your valuable suggestion for strengthening the quality of the manuscript.	2	1	Moderate English changes required	1	2	catal12030290_makarova	1
+Indeed, the liposomes are prepared by a conventional method. However, here and in our previous work [1] we use the term liposomal nanotraps to reflect the functional aspect of the liposomal action. In this context, the liposomes act as traps for bacterial toxins.	2	1	The authors used the term liposome nanotraps. The authors have used conventional liposome preparations. It is not clear why they are calling the preparations as the nanotraps.	1	2	cells11010166_perova	1
+We verified the total protein content of the supernatants by Coomassie blue staining, which was similar for all strains (Figure. 1, for referee inspection only). The experiments were performed using different bacterial supernatant batches and results were remarkably consistent. All the supernatants were collected at the exact same bacterial culture’s optical density to harvest the bacteria in a comparable state between batches and strains. In our previous work we have shown that cholesterol-dependent cytolysins (SLO, PLY) displayed different kinetics and dynamics of their hemolytic activity compared to SLS. As a result, the total hemolytic activities of individual streptococcal supernatants were not represented by a simple sum of activities (concentrations) of their individual toxins but displayed more complex time- and amount-dependent behavior: e.g. – the activity of PLY/SLO was prevalent at the initial times of incubation and at relatively high amounts (volume) of supernatants, whereas SLS activity fully developed only after initial lag period but was prevalent when relatively low amounts of supernatants were used in the assays. The relative quantifications of toxins between streptococcal species and strains were performed in a previous publication [1] and are referred to throughout the manuscript. We added a paragraph in the result section (line 215-222) that summarizes those points. Moreover, in our current experiments we aim not only at the neutralization of the whole hemolytic secretome of streptococcus (as described in [1]) but also include (putative) cytotoxic/cytostatic activities that might be carried out either by hemolysins (SLO/SLS) or other not yet identified toxins that display cytotoxic/cytostatic but no hemolytic activities. For these reasons, in our current experiments, we use specific cytotoxic/cytostatic activities of total supernatants derived from a toxicity assay displayed in (Figure. 1), instead of concentrations of individual (partly unknown) toxins. However, we agree with the reviewers that using volume units is confusing. We therefore, edited our manuscript to display the lethal dose (LD%) interpolated from results shown in Figure 1 instead. Using LD% units accounts for the batch variability.	2	1	The authors have used bacterial culture supernatants to examine the cytotoxic effect on the THP-1, Jurkat, and Raji cell lines. Quantitation fo the the culture medium is required (by measuring the total protein content, or by providing some quantitative indicator). Volume of the culture supernatant is mentioned in the results. It is not quantitative, and will vary from batch to bath of the bacterial culture.	1	2	cells11010166_perova	1
+While LDH-release or MTT assay are frequently used techniques and might be preferred by some investigators, other approaches assessing cell viability might be more popular by others, dependent on particular experimental settings of a particular study. Figure 2 (for referee inspection only) demonstrates that Alamar blue cell viability assay and Trypan blue live/dead quantification provide results that are identical to those obtained in the cell proliferation protocol used in our study. We believe that the latter protocol is the most suitable experimental approach for our study since it allows distinguishing between cell lysis, cytotoxisity and cytostatic effects (i.e. between cytolysins, cytotoxins and cytostatic agents) in a single experiment. The techniques and algorithms used by the CellDrop relies on accurate and unbiased measurements, akin to previously published techniques[2,3].	2	1	LDH-release assay of cytotoxicity or MTT assay of cell viability should have been used for measuring the cell death.	1	2	cells11010166_perova	1
+The focus of our study is to highlight that successful protection against the whole palette of streptococcal toxins can be achieved by using liposomal nanotraps and to show that the liposome requirements differs between bacterial species and between different types of immune cells. We agree that the SLO neutralization is not novel and simply confirm results from previous publications by us and others. In the current work we do not intend to reveal new neutralization mechanisms either for SLO or for SLS.	2	1	It is well-known that SLO binds to the cholesterol-containing lipid bilayer. Therefore, it is obvious that the cholesterol-containing liposomes would neutralise SLO present in the bacterial culture. Therefore, no new information is provided with these experiments.	1	2	cells11010166_perova	1
+Streptolysin S (SLS) is a small, non-immunogenic, peptide. This means that no commercial antibody against it is available and the peptide is too small for reliable mass spectrometry detection. It is heavily post-translationally modified and is the product of a complex operon. Its exact mode-of-action is still not yet fully clarified. In a previous publication, we were able to show that SLS is neutralized by phosphatidylcholine as well as sphingomyelin liposomes [1]. However, given the poor characterization of SLS, its unavailability from commercial providers and its extremely tedious purification, the mechanistic details of the SLS neutralization will require an extensive a project of its own and are beyond the scope of the current study.	2	1	In my opinion, differential inhibition of the SLS activity in GGS supernatant by the liposome preparations is interesting, and may provide new insights, if explored in more detail.	1	2	cells11010166_perova	1
+We used two controls in the protection experiments. In one control we challenged the cells with bacterial supernatant without adding liposomes, to determine that the baseline cytotoxicity is in line with the results displayed in the Figure 1. The second control consists of the immune cells without toxin or liposomes. This control represents 100% survival and allows us to normalize our survival data. We also tested the intrinsic toxicity of liposomes on their own to see if they did impede cell growth or were cytotoxic. At the concentrations used in current study the liposomes were not cytotoxic (data not shown). The following text was added to the material and method section: The data were normalized to a control incubated with PBS instead of bacterial supernatant (considered as 0% cell death). A control challenged by bacterial supernatant without liposomes was added for each assay to verify the expected cytotoxic activity 2.	2	1	Although the sensitivity of immune cells to GAS or GGS supernatants is shown in the first section of the results, were any controls used in the neutralization assays with liposomes?	1	2	cells11010166_perova	1
+We selected the specific supernatant volumes used to reach a similar lethal dose to compare the protection capability and efficiency of the liposomal nanotraps. However, as the immune cell lines have different sensitivities depending on the toxin profile of the tested strain, a similar lethal dose corresponds to different supernatant volumes. We used the results displayed in Figure 1 of the manuscript to determine which volume to use. We agree that the way we presented it can lead to confusion and we replaced the supernatant volume values by LD% values. The following text was added to the material and method section: The added supernatant volume was determined based on the toxicity assay results and was used either at saturating dose (lethal dose >90, LD>90) or at non-saturating dose (LD60-90) to study minor toxin activities. We agree that the distinction between saturating (LD>90) and non-saturating (LD60-90) was confusing. To clarify our process we did add clarifications throughout the results part and we added a panel in figure 2 to include results obtained after challenge of LD>90 of GGS 5804 supernatant.	2	1	In the material and methods section it is stated that the cells survival assays start with the addition of a fixed volume of supernatant but it is not clear to me the criteria for reflecting different amounts of supernatant in Figures 2 and 3. It should be explained more clearly. What is the reason for using different amounts of supernatant with cytolysins depending on its source or the type of cell line it is tested against in figures 2 and 3? This should be explained. Furthermore, it should be justified why this difference does not influence the comparison of the results with the different cell lines (THP1, Jurkat, Raji).	1	2	cells11010166_perova	1
+All the corrections are highlighted in yellow. Corrected the legend of Figure 4 (EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT).	2	1	Some errors in the legend of Figure 4, such as EM/PHYT/BHT, EM/PHYT is not presented in Figure 4.	1	2	cosmetics2030248_makarova	1
+Response: The errors were corrected in text on page 09. (EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT).	2	1	In Figure 4, the results is represented in three times or signal one or other times. Therefore, you must rewritten the legend of Figure 4.	1	2	cosmetics2030248_makarova	1
+All the corrections are highlighted in red. The “galenic” was mentioned in the article due to physician Claudio Galeno, which created the first creams formulations. Therefore we decided to change galenic to cosmetic.	2	1	We recommend “galenic” can be changed to herbaceous, because galenic is not usually to see for non-professional fields or general cosmetics reader.	1	2	cosmetics2030248_makarova	1
+Inserted parenthesis at line 132 (Tpeak = 48.17 °C).	2	1	Less a parenthesis at line 132.	1	2	cosmetics2030248_makarova	1
+Corrected at line 160 “Figureure” to “Figure”.	2	1	At line 160, “Figureure” should be “Figure.”	1	2	cosmetics2030248_makarova	1
+This abbreviation is widely use in Thermal analysis. We specified in the text (the extrapolated peak onset temperature (TonsetDSC).	2	1	At line 161 mentioned Tonset DCS, but does not explain what this word mean.	1	2	cosmetics2030248_makarova	1
+We removed the units of temperature from the Table.	2	1	In Table II, Tonset DCS is a units of temperature, it should be used comma instead of decimal point.	1	2	cosmetics2030248_makarova	1
+We changed “donate” to provide and the other word donor to provider.	2	1	At line 167, “donateis” is typically used to represent donate money, we proposed change to be “provide”, in additional, this sentence is not fluent at grammar.	1	2	cosmetics2030248_makarova	1
+We changed this sentence.	2	1	The sentence in line 168 is not fluent at grammar.	1	2	cosmetics2030248_makarova	1
+The sample´s name corrected (EM/BL) at line 189.	2	1	At line 189, it dors (does) not have sample named EBL, it needs to be check again.	1	2	cosmetics2030248_makarova	1
+We integrated all the problems and correct he errors.	2	1	Integration of all the above problems, this post Journal does not provide future applications of phytic acid in cosmetics and its title is mentioned in herbal lotion but the experiment did not use any herb emulsion formulation. For the antioxidant capacity of phytic acid only do the experiment of DPPH that has not more in-depth study. Lot of grammatical errors is found in sentences. So we suggested to big modifications, then returned for review.	1	2	cosmetics2030248_makarova	1
+We described these two points more clearly. The thermal analysis technique applies high temperatures in the samples due to this thermal stability of the emulsion associated with phytic acid decreased.	4	1	From the experimental results that phytic acid will decrease the thermal stability of the emulsion, but at line 220 mention that phytic acid has the effect of lipid peroxidation and therefore can be added as an anti-oxidant in the emulsion. So we recommend authors to describe these two points much clearer.	3	2	cosmetics2030248_makarova	1
+All the corrections are highlighted in green. We expanded the discussion of the results on pages 7, 10 and the conclusion on page 11 (item 4).	2	1	The conclusions reported are very short and poor. More discussion of the presented results is needed.	1	2	cosmetics2030248_makarova	1
+As mentioned in this paper, there are few studies related to the phytic acid in cosmetic products. However we added more references regarding about phytic acid antioxidant activity. References 24, 25 and 26.	2	1	The authors missed to report other authors’ studies which agree or disagree with their findings.	1	2	cosmetics2030248_makarova	1
+We mentioned in the text more information about incorporation of phytic acid into emulsion on Thermal Analysis results, DPPH results and conclusion.	2	1	What is the output of this study? What are the implications in the use of PA in galenic/cosmetic formulations?	1	2	cosmetics2030248_makarova	1
+Regarding with this queries we improved the discussion in the results on page 7. But we need to study more about.	2	1	What are the suggestions of the authors about the use of PA in Cosmetics based on their results? And what the parameters and the tricks that the formulators should be aware of while using PA in their products?	1	2	cosmetics2030248_makarova	1
+All the corrections are highlighted in yellow. Corrected the legend of Figure 4	2	1	Some errors in the legend of Figure 4, such as EM/PHYT/BHT, EM/PHYT is not presented in Figure 4.	1	2	cosmetics2030248_perova	1
+(EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT). The errors were corrected in text on page 09.	2	1	In Figure 4, the results is represented in three times or signal one or other times. Therefore, you must rewritten the legend of Figure 4.	1	2	cosmetics2030248_perova	1
+All the corrections are highlighted in red. The “galenic” was mentioned in the article due to physician Claudio Galeno, which created the first creams formulations. Therefore we decided to change galenic to cosmetic.	2	1	We recommend “galenic” can be changed to herbaceous, because galenic is not usually to see for non-professional fields or general cosmetics reader.	1	2	cosmetics2030248_perova	1
+Inserted parenthesis at line 132 (Tpeak = 48.17 °C).	2	1	Less a parenthesis at line 132.	1	2	cosmetics2030248_perova	1
+Corrected at line 160 “Figureure” to “Figure”.	2	1	At line 160, “Figureure” should be “Figure.”	1	2	cosmetics2030248_perova	1
+This abbreviation is widely use in Thermal analysis. We specified in the text (the extrapolated peak onset temperature (TonsetDSC).	2	1	At line 161 mentioned Tonset DCS, but does not explain what this word mean.	1	2	cosmetics2030248_perova	1
+We removed the units of temperature from the Table.	2	1	In Table II, Tonset DCS is a units of temperature, it should be used comma instead of decimal point.	1	2	cosmetics2030248_perova	1
+We changed “donate” to provide and the other word donor to provider.	2	1	At line 167, “donateis” is typically used to represent donate money, we proposed change to be “provide”, in additional, this sentence is not fluent at grammar.	1	2	cosmetics2030248_perova	1
+We changed this sentence.	2	1	The sentence in line 168 is not fluent at grammar.	1	2	cosmetics2030248_perova	1
+The sample´s name corrected (EM/BL) at line 189.	2	1	At line 189, it dors (does) not have sample named EBL, it needs to be check again.	1	2	cosmetics2030248_perova	1
+We integrated all the problems and correct he errors.	2	1	Integration of all the above problems, this post Journal does not provide future applications of phytic acid in cosmetics and its title is mentioned in herbal lotion but the experiment did not use any herb emulsion formulation. For the antioxidant capacity of phytic acid only do the experiment of DPPH that has not more in-depth study. Lot of grammatical errors is found in sentences. So we suggested to big modifications, then returned for review.	1	2	cosmetics2030248_perova	1
+We described these two points more clearly. The thermal analysis technique applies high temperatures in the samples due to this thermal stability of the emulsion associated with phytic acid decreased.	4	1	From the experimental results that phytic acid will decrease the thermal stability of the emulsion, but at line 220 mention that phytic acid has the effect of lipid peroxidation and therefore can be added as an anti-oxidant in the emulsion. So we recommend authors to describe these two points much clearer.	3	2	cosmetics2030248_perova	1
+All the corrections are highlighted in green. We expanded the discussion of the results on pages 7, 10 and the conclusion on page 11 (item 4).	2	1	The conclusions reported are very short and poor. More discussion of the presented results is needed.	1	2	cosmetics2030248_perova	1
+As mentioned in this paper, there are few studies related to the phytic acid in cosmetic products. However we added more references regarding about phytic acid antioxidant activity. References 24, 25 and 26.	2	1	The authors missed to report other authors’ studies which agree or disagree with their findings.	1	2	cosmetics2030248_perova	1
+We mentioned in the text more information about incorporation of phytic acid into emulsion on Thermal Analysis results, DPPH results and conclusion.	2	1	What is the output of this study? What are the implications in the use of PA in galenic/cosmetic formulations?	1	2	cosmetics2030248_perova	1
+Regarding with this queries we improved the discussion in the results on page 7. But we need to study more about.	2	1	What are the suggestions of the authors about the use of PA in Cosmetics based on their results? And what the parameters and the tricks that the formulators should be aware of while using PA in their products?	1	2	cosmetics2030248_perova	1
+Following the reviewer’s comments, we have now focused on one detailed analysis based on adequate tests and a discussion of the results. The revised version drops Descriptive statistics in old Table 2, rates of increase given in the old Table 3, correlations reported in old Table 4, the regressions in old Table 5, and pooled regressions of old Table 6. The discussion based on these Tables has consequently been omitted from the revision. The unit-root-test Tables have been moved from the Appendix to the text as Table 2, and in view of the evidence in favor of unit roots, rates of growth of GDP and government variables have been shown in Table 3 somewhat like the old Table 3. These are the most major changes.	2	1	I don’t really doubt the empirical findings of the paper (which I see as support for the hypothesis in Japan and possibly in Korea), but I dislike the approach of spending a lot of space on various, often pointless, tests. There should be one detailed analysis based on adequate tests, and a discussion about the results. After all, government expenditure is a political decision.	1	2	economies3040150_perova	1
+The authors are aware of the Johnson et al. paper (JME 2013) and other critiques of the PWT data. However, while the LCU numbers are appropriate for single-country analysis, these are not suitable for panel-data format which is a significant part of this study. Therefore, we use the PWT numbers throughout and hope that single-country analysis with these numbers is a reasonable approximation to the scenario indicated by LCU data.	2	1	An import question is why are the series for real GDP per capita are taken from PWT 7.0? These series are constructed to be used in cross-country studies, and they are not suitable for the analyses in the paper. See the paper by Johnson et al. (2009). This is a major flaw of the paper and the choice should be defended by showing that using the ‘standard’ real GDP series’ would give the same results. Or the GDP series, downloaded from World Development Indicators (local currency, constant prices), should be used.	1	2	economies3040150_perova	1
+The dependency variable has been removed from the analysis.	2	1	The dependency ratio has not been included mechanically in most papers, as it is this one. It is supposed to pick up changes in demand for government expenditure, such as schooling and pensions. The general idea is that these have increased government expenditure, both because per-capita schooling and pensions have expanded and because the dependency ratio has increased. In most of the countries studied, as is well-known, the dependency ratio declined during the period studied, while expenditure per person probably increased. Japan is most likely an exception; it should have entered the demographic transition much earlier than the other countries. There is thus no a priory no reason to expect the dependency ratio to play an important role in the Wagner hypothesis in general. It can thus be removed from the analysis, or moved to a footnote. If the author(s) wish to pursue the approach of testing other potentially relevant variables, see Shelton 2007.	1	2	economies3040150_perova	1
+Figure 1 has been redone with separate scales for logarithms of real GDP per capita and government share.	2	1	Figure 1 tends to hide co-movements between the variables by mixing of scales. I suggest the graphs are re-done using two y-axes and preferably logs.	1	2	economies3040150_perova	1
+The correlations of old Table 4, the static regressions of Table 5, and the discussion of endogeneity have been omitted.	2	1	In Table 5 we can get information about cointegration from the R2 and the t-values. Only Japan shows any promise, in all other cases R2 are very low. I would say they are suspiciously low for time series data. Are thy correct?	1	2	economies3040150_perova	1
+Since our data cover a long period of nearly 50 years, we are inclined to rely on Johansen tests although the number of data points is smaller than 100. The Gregory-Hansen test is done for Japan and Korea also just for completeness, and might be of some methodological interest since it does not show cointegration even for Japan and Korea. The reviewer’s observation has been acknowledged in note 2.	2	1	Gregory–Hansen is used when one fails to find cointegration with standard tests. There is no point in testing countries such as Japan and Korea, when a superior test shows that they are cointegrated.	1	2	economies3040150_perova	1
+We agree with the reviewer about the limited usefulness of MTAR in small samples and for Japan and Korea. However, it is included in the hope it would indicate whether lack of cointegration for Malaysia, Philippines, Singapore and Thailand might be due to asymmetric adjustment. The reviewer’s observation has been acknowledged in note 3.	2	1	The use of the MTAR model is dubious because of the small sample, and it should not be used when cointegration has been established. It is unlikely that there is enough information in the data to capture asymmetric effects and the standard procedure is to use MTAR when Engle-Granger fails to show cointegration.	1	2	economies3040150_perova	1
+We have highlighted the alternative hypothesis in Pedroni’s tests on Page 8. As Pedroni (1999) clearly points out, the alternative hypothesis rejects the null of no-cointegration across all cross-sections in both within- and between-dimension tests.	2	1	What is the alternative hypothesis of the Pedroni test, that there is cointegration in any country or or in all countries? This should be made clear.	1	2	economies3040150_perova	1
+The “provocative conclusion” reflected one author’s subjective perception and was stated as such. He shares the perspective noted by Bennett McCallum’s in his “Is the spurious regression problem spurious?” (Economics Letters, 2010). However, the segment cited by the reviewer has been deleted since it is a more general proposition and is not necessary for the limited analysis reported in our paper.	2	1	The authors provide a very provocative conclusion (see below), but it is not supported by the analyses in the paper. The single-country analysis would provide the same results if done carefully, and the panel data analyses are dubious both because of the small cross-section (six) and the heterogeneity of the sample. I don’t mind the use panel analyses they can be presented for illustrative purposes, noting the heterogeneity. However, the time series analyses should be done correctly. Moreover, it is quite obvious that in simple bivariate cases, graphs can be very informative. However, the author(s) do not manage to provide any useful information about the dependency ratio in the ‘old fashioned’ analyses; they do only in Table 5 when (unknowingly) testing for cointegrating using OLS.	1	2	economies3040150_perova	1
+We completely share the reviewer’s view that despite Biehl’s PF/FP essay, almost everyone follows the standard approach, and we do the same. Our observation is just a restatement of the proposition that our approach is traditional.	2	1	Despite Biehl’s (1998) insightful essay, almost all empirical research on the topic has interpreted the Wagnerian proposition As Ram (1998, Pp. 149-150) has explained, considerations of “nonstationarity” of the variables or “spuriousness” of the correlations or regression do not have much bearing on such tests.	1	2	economies3040150_perova	1
+(a) the plots in Figure 1 have been rescaled and now there are cross-overs in the plot for Japan, (b) years have been shown on the horizontal axis in Figure 1, (c) a sizable addition has been made toward the end of Section 3 to address the reviewer’s observations about possible reasons for lack of cointegration in most countries, and (d) a few minor editorial alterations have been made.	4	1	The x-axes in Figure 1 should have years.	3	2	economies3040150_perova	1
+Several new references, including those kindly mentioned by the reviewer, have been added, and there is a concise description of these in note 1.	2	1	List of references is scant and limited. It is necessary to add more recent works on the subject with alternative ways of testing for Wagner’s Law. Suggestions include “Testing the Validity of Wagner’s Law in Bolivia: A Cointegration and Causality Analysis with Disaggregated Data�� by A. Bojanic (2013); “Wagner’s Law and Italian Disaggregated Public Spending: Some Empirical Evidences” by C. Magazzino (2010); and “Panel Data, Cointegration, Causality and Wagner’s Law: Empirical Evidence from Chinese Provinces” by Narayan, P.K., et al. (2008).	1	2	economies3040150_perova	1
+Several formats in which Wagner’s hypothesis has been formulated and tested have been listed on page 5 of the revision. That paragraph includes six different approaches.	2	1	The paper only lists one way of testing for Wagner’s hypothesis (i.e., analyzing the relation between the share on GDP of government spending and real GDP per capita) when in fact there are many ways of looking at the hypothesis. The authors should include at least a couple of recent ways of testing this law, in addition to the traditional way of testing for it.	1	2	economies3040150_perova	1
+Real GDP per capita is a fairly standard proxy for the “level of development” in the context of Wagner’s hypothesis. It might not be appropriate to use nominal GDP per capita. For the government share, it seems better to take the current-price ratio which indicates what fraction of the current output is used for government activities. The deflators for GDP and government spending are different and “real” government-share may not be a good indicator of the resource-use by the government during the current year.	2	1	The variables used do not have the same units, which may clog the analysis of the paper. Specifically, real GDP per capita is measured in 2005 dollars while the share of government consumption on GDP is measured in current prices. Either both should be in real terms or both in current prices, but not as is currently the case.	1	2	economies3040150_perova	1
+The old Table 2 has been deleted. Those descriptive statistics were for the entire period. The growth rates in the new Table 3 are also for the entire period, and that has been indicated.	2	1	The descriptive stats in Table 2 reflect stats for the whole period? If so, the table should explicitly state this fact.	1	2	economies3040150_perova	1
+The redundant graphs for Philippines have been deleted.	2	1	Why are there 3 graphs for the Philippines in Figure 1? By the way, if the variables were measured in either current prices or utilizing the same base year, these graphs would likely be quite different (same goes for all statistical analysis, it is likely to vary when the two variables are measured the same way) 6.	1	2	economies3040150_perova	1
+The dependency-rate variable has been dropped.	2	1	What is the point of the dependency rate? The authors should expand on the need to add this variable and what exactly is it trying to capture. In the same vein, what do the authors mean by “however, it seems that the fall in young-dependency ratio was greater than the rise in the old-dependency rate” on p. 4?	1	2	economies3040150_perova	1
+The old Table 4 has been deleted along with the remark about p-values being taken from SAS.	2	1	In Table 4, delete the “generated by SAS” note.	1	2	economies3040150_perova	1
+We better specify the required items on agricultural labor productivity in the text. Sorry for the unclearness. The index is actually the measure of agricultural labor productivity, which is calculated by the agricultural output and labor. We now clarify this part with another way of expression.	2	1	Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to. Are the annual agricultural output data are measured in their nominal value (at current price) or in real value (at constant price)? If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions? Labor productivity needs to be measured using real value of agricultural output. Otherwise, it may reflect the changes in prices instead of productivity. This issue needs to be properly addressed.	1	2	economies4030013_makarova	1
+The Reviewer raises an interesting point that we overlooked previously. We take the starting year as 2000 is mainly because of the entry of WTO, which influences the pattern of agricultural production in China. We take the reviewer’s advice, to extend the time period until 2013, which is the latest available data on prefectural level.	2	1	Measurement issue: The authors rely on the Latent Growth Curve Modeling technique to study the trend growth of China’s agricultural labor productivity. It is not clear, however, why the authors rely on a dataset with only 10-year period to discuss labor productivity trend growth issue instead of using a dataset with longer time period if data is available. In addition, the authors may want to justify their approach by briefly discussing the advantage or shortcomings of their method comparing to other approaches in identifying the growth trend of China’s agricultural labor productivity.	1	2	economies4030013_makarova	1
+We now correct all the issues in citations and tables and figures in the text. Figure 3 (now is Figure 4), the vertical axis indicates the growth rate of agricultural labor productivity; we now put it in the note. In Appendix A1, the “absolute value” means real value; we now change it to the better expression. In Appendix A2, they are measured in real value at constant price; we now better specify it in the text.	2	1	Please be more specific. Appendix Figure A2: The figure presents the chain-linked index of output and input. It is not clear whether the output is measured in real value (at constant price) or nominal value (at current price).	1	2	economies4030013_makarova	1
+We would like to thank the reviewer’s scrutiny. We now specify this better in the text	2	1	One question that it is necessary to correct is the number of figures in the text.	1	2	economies4030013_makarova	1
+The agricultural output is deflated with the base year 2000. There is no price index at the prefectural level. For this reason we use the provincial price indices to deflate the nominal series. Hence, all the prefectures within each province are deflated by the same index. Although suboptimal, this helps to preserve the comparability in the two estimations. The reviewer rises a very useful point that we initially ignored. We now better specify the required items on agricultural labor productivity in the text.	2	1	Besides, the authors must define better the data how the agricultural labor productivity was calculated. The agricultural output is in current or constant prices? This could have strong implications in one case or another, especially in the first case. The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama. So, if the agricultural output is in current prices, the growing prices will affect deeply the results. It must be clearer.	1	2	economies4030013_makarova	1
+The reviewer is right. The first sector is a macro conception of agriculture. It indeed includes fishery. However, the data of output and employment are both collected on this level - the first sector. Hence, we assume the quotient of the two variables is a proxy to agriculture labor productivity. Meanwhile, they are the only available data on the prefectural level to calculate agricultural labor productivity. Therefore, we think this calculation is still helpful.	2	1	On the other hand, in the same line of text (pg. 4 line 3), the authors define the denominator of the agricultural labor productivity as the employed labors in the first sector. What does this first sector mean? Is it a synonymous of agricultural sector? If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access. In this case, it would be the primary sector labor productivity. The authors should also clarify if this sector included the forest products and labor.	1	2	economies4030013_makarova	1
+The Reviewer raises a very valuable point. We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.	2	1	The introduction is welldone, but it could be improved in some points. I have missed a review of the literature. The measurement of the agricultural labor productivity is an important theme in the literature, not only in the agricultural economics, but also in the economics literature. For example, there are two paper published in two important journals in economics recently that the authors omitted. These papers could be reinforcement about the relevance of this measurement of agricultural productivity: Gollin D, Lagakos D, Waugh ME (2014) Agricultural productivity differences across countries. American Economic Review 104(5):165–170; Gollin D, Lagakos D, Waugh ME (2014) The agricultural productivity gap. Quarterly Journal Economics 129(2): 939–993.	1	2	economies4030013_makarova	1
+We now specify this better in the text.	2	1	Other thing to improve the introduction would be writing the main objective of the paper. I think, this clear objective could improve the introduction about the target that the authors follow in this paper. In this version, this is relatively diffuse.	1	2	economies4030013_makarova	1
+The Reviewer raises a valuable point which we overlooked previously. In the section 3.3, we improve the understanding of the growth pattern of Chinese agricultural labor productivity by adding the estimations of beta and sigma convergence. The results of convergence estimation also echo our analysis of LGCM.	2	1	The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence. The authors wrote about these concepts, for example pg. 6 lines 22-23 and pg. 9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.	1	2	economies4030013_makarova	1
+Yes, the Reviewer is correct. Since we add three years into the dataset (following the suggestion from the other reviewer), the table context changes. We now improve this issue in the text.	2	1	Maybe, the footnote 5 could go to the table 2’s note.	1	2	economies4030013_makarova	1
+We improve this part in the text. The recent movements of the government in this agricultural reform are added. Since we add the data of recent three years, another break is observed in the curve. So we also introduce the macro policy of stimulus package.	2	1	One of the most interesting issues of the paper is the political change in 2004. Perhaps, more detailed reasons of the reform would enrich the causes to promote it. Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.	1	2	economies4030013_makarova	1
+We would like to thank the Reviewer to recommend us to develop this part. The Reviewer raises a very interesting point. However, the prefectural-level data in terms of farmland are only reported until 2007 in the China City Statistical Yearbook.	2	1	As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios. Maybe, some simple correlations between the growths of agricultural labor productivity vs land productivity and land-labor ratio could enrich this or future analysis.	1	2	economies4030013_makarova	1
+We better specify the required items on agricultural labor productivity in the text. Sorry for the unclearness. The index is actually the measure of agricultural labor productivity, which is calculated by the agricultural output and labor. We now clarify this part with another way of expression.	2	1	Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to. Are the annual agricultural output data are measured in their nominal value (at current price) or in real value (at constant price)? If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions? Labor productivity needs to be measured using real value of agricultural output. Otherwise, it may reflect the changes in prices instead of productivity. This issue needs to be properly addressed.	1	2	economies4030013_perova	1
+The Reviewer raises an interesting point that we overlooked previously. We take the starting year as 2000 is mainly because of the entry of WTO, which influences the pattern of agricultural production in China. We take the reviewer’s advice, to extend the time period until 2013, which is the latest available data on prefectural level.	2	1	Measurement issue: The authors rely on the Latent Growth Curve Modeling technique to study the trend growth of China’s agricultural labor productivity. It is not clear, however, why the authors rely on a dataset with only 10-year period to discuss labor productivity trend growth issue instead of using a dataset with longer time period if data is available. In addition, the authors may want to justify their approach by briefly discussing the advantage or shortcomings of their method comparing to other approaches in identifying the growth trend of China’s agricultural labor productivity.	1	2	economies4030013_perova	1
+We now correct all the issues in citations and tables and figures in the text. Figure 3 (now is Figure 4), the vertical axis indicates the growth rate of agricultural labor productivity; we now put it in the note. In Appendix A1, the “absolute value” means real value; we now change it to the better expression. In Appendix A2, they are measured in real value at constant price; we now better specify it in the text.	2	1	Please be more specific. Appendix Figure A2: The figure presents the chain-linked index of output and input. It is not clear whether the output is measured in real value (at constant price) or nominal value (at current price).	1	2	economies4030013_perova	1
+We would like to thank the reviewer’s scrutiny. We now specify this better in the text	2	1	One question that it is necessary to correct is the number of figures in the text.	1	2	economies4030013_perova	1
+The agricultural output is deflated with the base year 2000. There is no price index at the prefectural level. For this reason we use the provincial price indices to deflate the nominal series. Hence, all the prefectures within each province are deflated by the same index. Although suboptimal, this helps to preserve the comparability in the two estimations. The reviewer rises a very useful point that we initially ignored. We now better specify the required items on agricultural labor productivity in the text.	2	1	Besides, the authors must define better the data how the agricultural labor productivity was calculated. The agricultural output is in current or constant prices? This could have strong implications in one case or another, especially in the first case. The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama. So, if the agricultural output is in current prices, the growing prices will affect deeply the results. It must be clearer.	1	2	economies4030013_perova	1
+The reviewer is right. The first sector is a macro conception of agriculture. It indeed includes fishery. However, the data of output and employment are both collected on this level - the first sector. Hence, we assume the quotient of the two variables is a proxy to agriculture labor productivity. Meanwhile, they are the only available data on the prefectural level to calculate agricultural labor productivity. Therefore, we think this calculation is still helpful.	2	1	On the other hand, in the same line of text (pg. 4 line 3), the authors define the denominator of the agricultural labor productivity as the employed labors in the first sector. What does this first sector mean? Is it a synonymous of agricultural sector? If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access. In this case, it would be the primary sector labor productivity. The authors should also clarify if this sector included the forest products and labor.	1	2	economies4030013_perova	1
+The Reviewer raises a very valuable point. We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.	2	1	The introduction is welldone, but it could be improved in some points. I have missed a review of the literature. The measurement of the agricultural labor productivity is an important theme in the literature, not only in the agricultural economics, but also in the economics literature. For example, there are two paper published in two important journals in economics recently that the authors omitted. These papers could be reinforcement about the relevance of this measurement of agricultural productivity: Gollin D, Lagakos D, Waugh ME (2014) Agricultural productivity differences across countries. American Economic Review 104(5):165–170; Gollin D, Lagakos D, Waugh ME (2014) The agricultural productivity gap. Quarterly Journal Economics 129(2): 939–993.	1	2	economies4030013_perova	1
+We now specify this better in the text.	2	1	Other thing to improve the introduction would be writing the main objective of the paper. I think, this clear objective could improve the introduction about the target that the authors follow in this paper. In this version, this is relatively diffuse.	1	2	economies4030013_perova	1
+The Reviewer raises a valuable point which we overlooked previously. In the section 3.3, we improve the understanding of the growth pattern of Chinese agricultural labor productivity by adding the estimations of beta and sigma convergence. The results of convergence estimation also echo our analysis of LGCM.	2	1	The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence. The authors wrote about these concepts, for example pg. 6 lines 22-23 and pg. 9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.	1	2	economies4030013_perova	1
+Yes, the Reviewer is correct. Since we add three years into the dataset (following the suggestion from the other reviewer), the table context changes. We now improve this issue in the text.	2	1	Maybe, the footnote 5 could go to the table 2’s note.	1	2	economies4030013_perova	1
+We improve this part in the text. The recent movements of the government in this agricultural reform are added. Since we add the data of recent three years, another break is observed in the curve. So we also introduce the macro policy of stimulus package.	2	1	One of the most interesting issues of the paper is the political change in 2004. Perhaps, more detailed reasons of the reform would enrich the causes to promote it. Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.	1	2	economies4030013_perova	1
+We would like to thank the Reviewer to recommend us to develop this part. The Reviewer raises a very interesting point. However, the prefectural-level data in terms of farmland are only reported until 2007 in the China City Statistical Yearbook.	2	1	As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios. Maybe, some simple correlations between the growths of agricultural labor productivity vs land productivity and land-labor ratio could enrich this or future analysis.	1	2	economies4030013_perova	1
+According to reviewer’s comment, we added Equation (14), and Tables 3 and 4 on the number of additions for the VVC inverse transforms.	2	1	Please add information on the number of additions for different transform types and sizes in original and proposed variants.	1	2	electronics11050760_makarova	1
+According to the reviewer’s comment, we modified and added some keywords as follows: We replaced “Keywords: VVC; HEVC; video coding; transform; computational complexity” With “Keywords: VVC (Versatile Video Coding), HEVC (High Efficiency Video Coding, Linear Inverse Transform, computational complexity, DCT (Discrete Cosine Transform), Discrete Sine Transform (DST), BD-rate”	2	1	It is better that they replace them with other keywords to increase the reach of the manuscript. Keywords : We suggest that the authors should replace keywords such as “video coding” and “transform” because these keywords are already found in the article title.	1	2	electronics11050760_makarova	1
+According to the reviewer's comments, we updated the last paragraph of Section 1 as follows: We replaced “Section 2 presents a proposed inverse transform using linearity to reduce the computational complexity.” With “In Section 2.1, we introduce the transforms used in VVC. And we propose a fast inverse transform using linearity to reduce the computational complexity in Section 2.2.” Comment) Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	2	1	Introduction: The authors should add the main contribution s briefly at the end of the introduction.	1	2	electronics11050760_makarova	1
+According to the reviewer’s comment, we divided Section 2 as follows: 2. VVC Transforms and Proposed Method 2.1. Introduction to DCT-II, DST-VII, and DCT-VIII 2.2. Propose Fast Inverse Transform Using Linearity Comment) The authors should add a section where they discuss comparing their results with those of existing research.	2	1	Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	1	2	electronics11050760_makarova	1
+According to the reviewer’s comment, we included the following sentence in the end of 3. Experimental Results section on page 11: ~~ “Fast encoding methods only in the encoder side were proposed to reduce the encoding complexity of VVC but all the fast encoding methods increased the BD-rates [29][30][31] in terms of the bit-rate reduction so that the proposed inverse transform using linearity in the decoder side differs from those approaches in that it keeps the BD-rate in VVC while reducing the decoding complexity. If the proposed inverse transform is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. Therefore, the proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.” ~~ And three new references [29],[30], and [31] in Reference section were included.	2	1	There is a lot of recent research out there that can be used for comparison. As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.	1	2	electronics11050760_makarova	1
+According to the reviewer’s comment, we added the future directions in the conclusion section as follows: “The proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.”	2	1	Conclusions: Future directions should be added to the conclusion section.	1	2	electronics11050760_makarova	1
+High Efficient Video Coding (HEVC), Standard ITU-T Recommendation H.265 and ISO/IEC 23008-2, Apr. 2013. Sullivan, G. J.; Ohm, J.; Han, W.; Wiegand, T. Overview of the High Efficiency Video Coding (HEVC) Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2012, vol. 22, pp. 1649-1668, doi: 10.1109/TCSVT.2012.2221191. Bross, B.; Chen, J.; Liu, S.; Wang, Y.-K. Versatile Video Coding (Draft 10). In Proceedings of the 19th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Teleconference (Online), 22 June-1 July 2020; 6. Bross, B.; Wang, Y.-K; Ye, Y,; Liu, S.; Sullivan, G. J.; Ohm, J. Overview of the Versatile Video Coding (VVC) Standard and its Applications, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol. 31, pp. 3736-3764, doi: 10.1109/TCSVT.2021.3101953 7. Karhunen, K. Über Lineare Methoden in der Wahrscheinlichkeitsrechnung, Soumalainen Tiedeakatemia, 1947; pp. 1-79.	2	1	The names of the researchers must follow the style of the journal format.	1	2	electronics11050760_makarova	1
+Thank you for pointing out our mistake. According to the reviewer’s comment, we completely updated the list of references as follows: 1. Advanced Video Coding (AVC), Standard ITU-T Recommendation H.264 and ISO/IEC 14496-10, May 2003. Wiegand, T.; Sullivan, G. J.; Bjontegaard, G.; Luthra, A. Overview of the H.264/AVC Video Coding Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2003, vol. 13, pp. 560-576, doi: 10.1109/TCSVT.2003.815165..	2	1	Some links do not work in the reference list like [22] … etc.	1	2	electronics11050760_makarova	1
+According to the reviewer’s comment, the revised manuscript was reviewed and substantially modified by an English native speaker.	2	1	This article requires extensive proofreading. Authors should check the entire article to remove all extensive mistakes (grammatical and typos) and to improve English writing quality.	1	2	electronics11050760_makarova	1
+According to reviewer’s comment, we the following sentences at the end of the Introduction. “In this paper, we analyze the number of multiplications of the existing fast transform methods in the VVC standard, and we propose a new fast inverse transform using the number of non-zero coefficients based on linearity to reduce the number of multiplications.” Comment 2) Discussion Section: The authors did not respond accurately to this comment.	4	1	The authors should add the main contributions briefly at the end of the introduction. Introduction Section: This comment still requires a response.	3	2	electronics11050760_makarova	1
+According to reviewer’s comment, we added Discussion Section. Discussion The previously proposed fast methods were mainly addressed to reduce complexity in the video encoder with the BD-rate loss. In [29], a fast intra mode decision algorithm was proposed and the result showed the encoding time savings of 51%~53% with BD-rate loss of 0.93%~1.08%. A low-complexity CTU (Coding Tree Unit) partition structure decision and fast intra mode decision were proposed in [30] and showed the average encoding time saving of 63% with the BD-rate loss of 1.93%. The fast encoders for video coding reduce only the encoder complexity but the BD-rates were always increased without decreasing the decoder complexity. However, the proposed fast inverse transform is different from the fast encoders in that it reduces the complexity in both the encoder and decoder while maintaining the BD- rate of the VVC standard. In the RA configuration, the proposed method reduces the average encoding and decoding times by approximately (4, 10) %, respectively, while maintaining average BD-rates. If the proposed inverse transform using the number of non-zero coefficients is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. This fact can be demerit. However, the proposed method can be considered in the next-generation video coding standards because it achieves decoding run-time saving, while maintaining average BD-rate. In addition to that, the proposed method is more effective in high QP value than in low QP value, because the higher the QP value is, the fewer no-zero coefficients there are.	4	1	The authors should add a section where they discuss comparing their results with those of existing research. Also, they should clarify the limitations of the proposed method. Discussion Section: The authors did not respond accurately to this comment.	3	2	electronics11050760_makarova	1
+We cannot find them.	4	1	Figures and Tables: All figures and tables are shown before they are used in the text.	3	2	electronics11050760_makarova	1
+According to reviewer’s comment, we added Equation (14), and Tables 3 and 4 on the number of additions for the VVC inverse transforms.	2	1	Additions also affect complexity. Please add information on the number of additions for different transform types and sizes in original and proposed variants.	1	2	electronics11050760_perova	1
+According to the reviewer’s comment, we modified and added some keywords as follows: We replaced “Keywords: VVC; HEVC; video coding; transform; computational complexity” With “Keywords: VVC (Versatile Video Coding), HEVC (High Efficiency Video Coding, Linear Inverse Transform, computational complexity, DCT (Discrete Cosine Transform), Discrete Sine Transform (DST), BD-rate” Comment) Introduction: The authors should add the main contribution s briefly at the end of the introduction.	2	1	Keywords: We suggest that the authors should replace keywords such as “video coding” and “transform” because these keywords are already found in the article title. It is better that they replace them with other keywords to increase the reach of the manuscript.	1	2	electronics11050760_perova	1
+According to the reviewer's comments, we updated the last paragraph of Section 1 as follows: We replaced “Section 2 presents a proposed inverse transform using linearity to reduce the computational complexity.” With “In Section 2.1, we introduce the transforms used in VVC. And we propose a fast inverse transform using linearity to reduce the computational complexity in Section 2.2.” Comment) Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	2	1	Introduction Section:	1	2	electronics11050760_perova	1
+According to the reviewer’s comment, we divided Section 2 as follows: 2. VVC Transforms and Proposed Method 2.1. Introduction to DCT-II, DST-VII, and DCT-VIII 2.2. Propose Fast Inverse Transform Using Linearity Comment) The authors should add a section where they discuss comparing their results with those of existing research.	2	1	Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	1	2	electronics11050760_perova	1
+According to the reviewer’s comment, we included the following sentence in the end of 3. Experimental Results section on page 11: ~~ “Fast encoding methods only in the encoder side were proposed to reduce the encoding complexity of VVC but all the fast encoding methods increased the BD-rates [29][30][31] in terms of the bit-rate reduction so that the proposed inverse transform using linearity in the decoder side differs from those approaches in that it keeps the BD-rate in VVC while reducing the decoding complexity. If the proposed inverse transform is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. Therefore, the proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.” ~~ And three new references [29],[30], and [31] in Reference section were included.	2	1	Discussion Section: There is a lot of recent research out there that can be used for comparison. As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.	1	2	electronics11050760_perova	1
+According to the reviewer’s comment, we added the future directions in the conclusion section as follows: “The proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.”	2	1	Conclusions: Future directions should be added to the conclusion section.	1	2	electronics11050760_perova	1
+Thank you for pointing out our mistake. According to the reviewer’s comment, we completely updated the list of references as follows: 1. Advanced Video Coding (AVC), Standard ITU-T Recommendation H.264 and ISO/IEC 14496-10, May 2003. 2. Wiegand, T.; Sullivan, G. J.; Bjontegaard, G.; Luthra, A. Overview of the H.264/AVC Video Coding Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2003, vol. 13, pp. 560-576, doi: 10.1109/TCSVT.2003.815165.. 3. High Efficient Video Coding (HEVC), Standard ITU-T Recommendation H.265 and ISO/IEC 23008-2, Apr. 2013. 4. Sullivan, G. J.; Ohm, J.; Han, W.; Wiegand, T. Overview of the High Efficiency Video Coding (HEVC) Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2012, vol. 22, pp. 1649-1668, doi: 10.1109/TCSVT.2012.2221191. 5. Bross, B.; Chen, J.; Liu, S.; Wang, Y.-K. Versatile Video Coding (Draft 10). In Proceedings of the 19th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Teleconference (Online), 22 June-1 July 2020; 6. Bross, B.; Wang, Y.-K; Ye, Y,; Liu, S.; Sullivan, G. J.; Ohm, J. Overview of the Versatile Video Coding (VVC) Standard and its Applications, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol. 31, pp. 3736-3764, doi: 10.1109/TCSVT.2021.3101953 7. Karhunen, K. Über Lineare Methoden in der Wahrscheinlichkeitsrechnung, Soumalainen Tiedeakatemia, 1947; pp. 1-79. 8. Ahmed, N.; Natarajan, T.; Rao, K. R. Discrete Cosine Transform, In IEEE Transactions on Computers, 1974, vol. C-23, pp. 90-93, doi: 10.1109/T-C.1974.223784. 9. Rose, K.; Heiman, A.; Dinstein, I. DCT/DST Alternate-Transform Image Coding, In IEEE Transactions on Communications, 1990, vol. 38, pp. 94-101, doi: 10.1109/26.46533. 10. Zhao, X.; Chen, J.; Karczewicz, M.; Zhang, L.; Li, X.; Chien, W. -J. Enhanced Multiple Transform for Video Coding, 2016 Data Compression Conference (DCC), 2016, pp. 73-82, doi: 10.1109/DCC.2016.9. 11. Han, J.; Saxena, A.; Melkote, V.; Rose, K. Jointly Optimized Spatial Prediction and Block Transform for Video and Image Coding, In IEEE Transactions on Image Processing, 2012, vol. 21, pp. 1874-1884, doi: 10.1109/TIP.2011.2169976. 12. Budagavi, M.; Fuldseth, A.; Bjøntegaard, G.; Sze, V.; Sadafale, M. Core Transform Design in the High Efficiency Video Coding (HEVC) Standard, In IEEE Journal of Selected Topics in Signal Processing, 2013, vol. 7, pp. 1029-1041, doi: 10.1109/JSTSP.2013.2270429. 13. Zhao, X.; Chen, J.; Karczewicz, M.; Said, A.; Seregin, V. Joint Separable and Non-Separable Transforms for Next-Generation Video Coding, In IEEE Transactions on Image Processing, 2018, vol. 27, no. 5, pp. 2514-2525, doi: 10.1109/TIP.2018.2802202. 14. Zhao, X.; Chen, J.; Said, A.; Seregin, V.; Egilmez, H. E.; Karczewicz, M. NSST: Non-separable secondary transforms for next generation video coding, 2016 Picture Coding Symposium (PCS), 2016, pp. 1-5, doi: 10.1109/PCS.2016.7906344.	2	1	References List: The list of references is recent, and all references are related to the research topic but it is not sufficient for this study. The names of the researchers must follow the style of the journal format. The double quotation should be omitted from the research titles in the list of references. Some search names in the reference list begin an uppercase letter for each word (such as [4], [5] ... etc.) and others use only an uppercase letter in the first word (such as [2], [9] … etc. ), authors should standardize style. All journal names should be italic. Some references do not contain enough information such as [16], [18] … etc. Some links do not work in the reference list like [22] … etc. The list of references requires an extensive check.	1	2	electronics11050760_perova	1
+According to reviewer’s comment, we the following sentences at the end of the Introduction. “In this paper, we analyze the number of multiplications of the existing fast transform methods in the VVC standard, and we propose a new fast inverse transform using the number of non-zero coefficients based on linearity to reduce the number of multiplications.” Comment 2) Discussion Section: The authors did not respond accurately to this comment.	4	1	The authors should add the main contributions briefly at the end of the introduction. Introduction Section: This comment still requires a response.	3	2	electronics11050760_perova	1
+4. According to reviewer’s comment, we added Discussion Section. Discussion The previously proposed fast methods were mainly addressed to reduce complexity in the video encoder with the BD-rate loss. In [29], a fast intra mode decision algorithm was proposed and the result showed the encoding time savings of 51%~53% with BD-rate loss of 0.93%~1.08%. A low-complexity CTU (Coding Tree Unit) partition structure decision and fast intra mode decision were proposed in [30] and showed the average encoding time saving of 63% with the BD-rate loss of 1.93%. The fast encoders for video coding reduce only the encoder complexity but the BD-rates were always increased without decreasing the decoder complexity. However, the proposed fast inverse transform is different from the fast encoders in that it reduces the complexity in both the encoder and decoder while maintaining the BD- rate of the VVC standard. In the RA configuration, the proposed method reduces the average encoding and decoding times by approximately (4, 10) %, respectively, while maintaining average BD-rates. If the proposed inverse transform using the number of non-zero coefficients is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method.	4	1	The authors should add a section where they discuss comparing their results with those of existing research. Also, they should clarify the limitations of the proposed method. Discussion Section: The authors did not respond accurately to this comment.	3	2	electronics11050760_perova	1
+We cannot find them.	4	1	Figures and Tables: All figures and tables are shown before they are used in the text.	3	2	electronics11050760_perova	1
+The diesel generator equal to the rating of base load is kept as backup to avoid total black out situation of the system.	2	1	In the proposed system solar PV, wind generator and battery are the main generating system. As the proposed system is in isolated mode, how continuity of power supply be maintained?	1	2	en15031006_perova	1
+According to your suggestion, we have improved the quality of Figures as can see Figure 1 and Figure2.	2	1	The quality of the figures needs to be improved.	1	2	en15031006_perova	1
+According to your suggestion, we have modified the Equations 4, 9 and 12 in the revised version of the manuscript as fellows.	2	1	Equations 4, 9, and 12 include some corrections and need to be modified.	1	2	en15031006_perova	1
+According to your suggestion the baseload is added in the result section as follows: In addition, to further validate the proposed system, the appliance schedule pattern for Monday is drawn in Figure 10 and Figure 9 represents the appliance weightage given by consumers for Monday. In addition, Figure 8 represents the power generated graph for the same day. The baseload is calculated as 1000 watts for each house to keep basic appliances turn ON uninterruptedly. As there are 5 houses in the community the total base power for the system is 5000 watts. The excess power is utilize to turn ON the schedulable appliances on Monday.	2	1	It could be better to include base load ratings in the results section (Fig: 10)	1	2	en15031006_perova	1
+As Sizing of the grid system is not the main goal of the article, but as per your suggestion, it is briefly discuss in the subsections of Mathematical modelling of the Energy sources. Both solar and wind systems are designed for output power equal to the consumer’s load whose annual load curve peak is maximum out of all the consumer’s in the community. Battery storage systems have the capability to supply uninterrupted power equal to the base load of the system up to 6 hours. And diesel generator equal to the rating of system’s base load is kept as a backup to avoid total black out situation. The designed system is not the time-varying tariff, therefore the overall system is designed to treat each consumer of the community equally. In time-varying tariff based system, the sizing of the system is the main objective as to accommodate the wealthy consumers of the community.	2	1	Sizing of renewable energy sources and battery systems were not properly explored.	1	2	en15031006_perova	1
+According to your suggestion the consumer comfortability is discussed from line 144-160. In this work, a DSM algorithm is proposed by using renewable resources PV and wind turbines for those houses that are located far away from the main city.	2	1	Please discuss how consumers’ comfortability is marinated in the proposed method.	1	2	en15031006_perova	1
+According to your suggestion, in the revised version of the manuscript, the unnecessary use of subjective pronoun is removed and overall writing of the paper is improved.	2	1	Authors should avoid using the Subjective pronoun “ we” in academic works.	1	2	en15031006_perova	1
+Thank you for your comment. Due to the lack of real time datasets, we have used the publically available datasets for the verification of the proposed system.	2	1	In such a study an experimental case must be presented in a specified region.	1	2	en15031006_perova	1
+Battery storage bank is included in the system to provide the baseload power in the time when renewable sources are not suffice to meet the baseload power demand. Diesel generators equal to the rating of base load is kept as backup to avoid total black out in worse conditions. Our objective is to schedule and shift the controllable appliances by estimating hourly power generation and considering consumer demand as a variable function. The appliances are prioritized based on their need and demand to achieve optimum energy utilization. The main contribution of this paper include: A demand side management algorithm is proposed to fulfil the energy gap between generation and consumer’s demand for standalone renewable energy system. K-mean clustering is used to make clusters of the data based on two factors: probability of turning ON a specific appliance at time t and priority number given by consumer to that specific appliance. Linear integer programming is used to schedule the appliances clusters based on the available power and state of charge of the battery system. According to your suggestion, the novelty of the proposed work has been made more clear and understandable for readers in the introduction section as follows: In this work, a DSM algorithm is proposed by using renewable resources PV and wind turbines for those houses that are located far away from the main city.	2	1	The authors should carefully distinguish the new contributions of their work from the new existing studies.for example: DOI: 10.1016/j.est.2020.101221	1	2	en15031006_perova	1
+According to your suggestion, we have modified the literature review and included some latest references i.e., [22][29][30][32]. In addition, we organized the literature review in more comprehensive way.	2	1	A comprehensive deeper literature review is necessary to address the research issue. Also, authors need to provide a literature survey in an organized way.-	1	2	en15031006_perova	1
+According to your suggestion, we have improved the Figures as can be seen in the revised version of the manuscript.	2	1	All the figures need to be improved.	1	2	en15031006_perova	1
+We have taken your advice and changed statistical methods in the manuscript.	2	1	English language should be improved.	1	2	fermentation8050212_makarova	1
+We have improved English language.	2	1	"Sometimes the document is different to follow due to English issues. Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology. ""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH. Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al. (2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”."	1	2	fermentation8050212_makarova	1
+FRBE is water-soluble fermented rice bran extracts.	2	1	Explain what FRBE is. It appears for the first time on Page 1, Line 17; Response: Thanks for your comment.	1	2	fermentation8050212_makarova	1
+The hot water extraction conditions was adopt according to previous methods (Dong et al., 2021). Dong, C.L. ; Liu, N.; Wang, Y.; Song, M.; Wang, R.F. ; Yang, Y.P. ; An, X.P. Study on Antioxidant Activity of Fermented Rice Bran Water Extract. Feed Industry 2021, 42(24), 8-13. DOI:10.13302/j.cnki.fi.2021.24.002.	2	1	The hot water extraction conditions were tested before?	1	2	fermentation8050212_makarova	1
+24hpf means 24 hours post fertilization.	2	1	What do the authors mean by 24hpf (Page 4, Line 148)?	1	2	fermentation8050212_makarova	1
+AO solution is acridine orange.	2	1	Explain what AO solution is. It appears on Page 4, Line 162; Response: Thanks for your comment.	1	2	fermentation8050212_makarova	1
+; Qi, J.W. Humates added to feed stimulate the microbial growth(Huck et al., 1991). Sodium humate added to fermentation substrate improved the growth of bacteria (Liu et al., 2014, Shi et al., 2016). The role of sodium humate is fermentation synergist. Huck, T.A., N. Porter and M.E. Bushell, 1991. Effect of humates on microbial activity. Gen. Microbiol. 137: 2321-2329. Liu, N.; An, X.P. ; Tong, B. S.; Chen, D. Y.; Yu, C.Q. Study on the effect of sodium humate on solid fermentation of lactobacillus plantarum. Chinese Journal of Animal Science 2014, 50(05),79-82. Shi, J.X. ; Qi, J.W; An, X.P. ; Liu, N.; Luo, X.G; Chen, D.Y. The influence of sodium humate on fermentation characteristics of yeast. Feed Industry, 2016, 37(04), 44-47.	2	1	Explain the role of sodium humate.	1	2	fermentation8050212_makarova	1
+It is my mistake to write the water-soluble polysaccharide content of FRBE and RBE on the wrong sides. The water-soluble polysaccharide content values have been corrected.	2	1	Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.	1	2	fermentation8050212_makarova	1
+We have taken your advice to change the number of decimal places.	2	1	Table 1: The number of decimal places of some constituents is excessive. Example: for polysaccharide content, I suggest writing 552+3 for RBE.	1	2	fermentation8050212_makarova	1
+Phenolic acids belongs to polyphenols. The increasing of content of water-soluble phenolic acids in FRBE may be attributed to the degradation of rice bran cell wall during fermentation.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?	1	2	fermentation8050212_makarova	1
+Fig.3a and 3b showing different groups of LPO rate.	2	1	Page 9, Line 280: Is Fig. 3b showing different groups of LPO rate, or is Fig.3a?	1	2	fermentation8050212_makarova	1
+The 73.85 and 77.00 values are correct.	2	1	Page 10, Lines 306 and 307: check the 73.85 and 77.00 values. They seem interchanged with each other.	1	2	fermentation8050212_makarova	1
+We have rewrited the first sentence in Conclusions. In summary, the DPPH radical radical scavenging abilities of rice bran water-soluble extracts and the content of bioactive constituents were considerably altered by mixed bacteria solid-state fermentation.	2	1	Rewrite the first sentence in Conclusions.	1	2	fermentation8050212_makarova	1
+I have changed.	2	1	Page 8, Line 261: Change “FRB” to “FRBE”.	1	2	fermentation8050212_makarova	1
+The abbreviated form of some journal titles have been corrected to full form taking into account the rules of the journal.	2	1	In the reference list, write the names of the microorganisms in italic. Some journal titles are registered in abbreviated form, and others have a full name. Standardize, taking into account the rules of the journal.	1	2	fermentation8050212_makarova	1
+The statistical analysis were conducted as follows: All experiments were conducted in triplicate. The UNIVARIATE procedure of SAS was used to test the normality of the data before any further analyses were carried out. Data were subjected to ANOVA using the MIXED procedure of SAS (SAS 9.2, SAS Institute Inc., Cary, NC). Mean ± standard error (S.E.) were reported, and means were separated by LSD multiple comparisons. Differences were accepted as statistically significance at P < 0.05.	2	1	The main comment addressing the article improvement is the methodology applied for statistical analysis. The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied. Later one-factor analysis of variance should carry out accordingly. The application of the Duncan test should be justified because it is less sensitive in this kind of study.	1	2	fermentation8050212_makarova	1
+Thanks for your comment. We have enhanced clarity and detail of the image for easy reading.	2	1	Figures 2a, 3a and 4a include photos whose resolution should be improved.	1	2	fermentation8050212_makarova	1
+We have taken your advice and changed statistical methods in the manuscript.	2	1	please include a more specific section about the statistical methods.	1	2	fermentation8050212_perova	1
+We have improved English language.	2	1	"Sometimes the document is different to follow due to English issues. Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology. ""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH. Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al. (2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”."	1	2	fermentation8050212_perova	1
+FRBE is water-soluble fermented rice bran extracts.	2	1	Explain what FRBE is. It appears for the first time on Page 1, Line 17; Response: Thanks for your comment.	1	2	fermentation8050212_perova	1
+The hot water extraction conditions was adopt according to previous methods (Dong et al., 2021). Dong, C.L. ; Liu, N.; Wang, Y.; Song, M.; Wang, R.F. ; Yang, Y.P. ; An, X.P. ; Qi, J.W. Study on Antioxidant Activity of Fermented Rice Bran Water Extract. Feed Industry 2021, 42(24), 8-13. DOI:10.13302/j.cnki.fi.2021.24.002.	2	1	The hot water extraction conditions were tested before?	1	2	fermentation8050212_perova	1
+24hpf means 24 hours post fertilization.	2	1	What do the authors mean by 24hpf (Page 4, Line 148)?	1	2	fermentation8050212_perova	1
+AO solution is acridine orange.	2	1	Explain what AO solution is. It appears on Page 4, Line 162; Response: Thanks for your comment.	1	2	fermentation8050212_perova	1
+Humates added to feed stimulate the microbial growth(Huck et al., 1991). Sodium humate added to fermentation substrate improved the growth of bacteria (Liu et al., 2014, Shi et al., 2016). The role of sodium humate is fermentation synergist.	2	1	Explain the role of sodium humate.	1	2	fermentation8050212_perova	1
+It is my mistake to write the water-soluble polysaccharide content of FRBE and RBE on the wrong sides. The water-soluble polysaccharide content values have been corrected.	2	1	Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.	1	2	fermentation8050212_perova	1
+We have taken your advice to change the number of decimal places.	2	1	Table 1: The number of decimal places of some constituents is excessive. Example: for polysaccharide content, I suggest writing 552+3 for RBE.	1	2	fermentation8050212_perova	1
+Phenolic acids belongs to polyphenols. The increasing of content of water-soluble phenolic acids in FRBE may be attributed to the degradation of rice bran cell wall during fermentation.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?	1	2	fermentation8050212_perova	1
+Fig.3a and 3b showing different groups of LPO rate.	2	1	Page 9, Line 280: Is Fig. 3b showing different groups of LPO rate, or is Fig.3a?	1	2	fermentation8050212_perova	1
+The 73.85 and 77.00 values are correct.	2	1	Page 10, Lines 306 and 307: check the 73.85 and 77.00 values. They seem interchanged with each other.	1	2	fermentation8050212_perova	1
+We have rewrited the first sentence in Conclusions. In summary, the DPPH radical radical scavenging abilities of rice bran water-soluble extracts and the content of bioactive constituents were considerably altered by mixed bacteria solid-state fermentation.	2	1	Rewrite the first sentence in Conclusions.	1	2	fermentation8050212_perova	1
+The “oscillated for 10 minutes” means “shaked for 10 minutes”.	2	1	Page 3, Line 124: What do you mean “oscillated for 10 minutes”? Is mixing for 10 minutes?	1	2	fermentation8050212_perova	1
+I have changed.	2	1	Page 8, Line 261: Change “FRB” to “FRBE”.	1	2	fermentation8050212_perova	1
+The abbreviated form of some journal titles have been corrected to full form taking into account the rules of the journal.	2	1	In the reference list, write the names of the microorganisms in italic. Some journal titles are registered in abbreviated form, and others have a full name. Standardize, taking into account the rules of the journal.	1	2	fermentation8050212_perova	1
+The statistical analysis were conducted as follows: All experiments were conducted in triplicate. The UNIVARIATE procedure of SAS was used to test the normality of the data before any further analyses were carried out. Data were subjected to ANOVA using the MIXED procedure of SAS (SAS 9.2, SAS Institute Inc., Cary, NC). Mean ± standard error (S.E.) were reported, and means were separated by LSD multiple comparisons. Differences were accepted as statistically significance at P < 0.05.	2	1	The main comment addressing the article improvement is the methodology applied for statistical analysis. The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied. Later one-factor analysis of variance should carry out accordingly. The application of the Duncan test should be justified because it is less sensitive in this kind of study.	1	2	fermentation8050212_perova	1
+Thanks for your comment. We have enhanced clarity and detail of the image for easy reading.	2	1	Figures 2a, 3a and 4a include photos whose resolution should be improved.	1	2	fermentation8050212_perova	1
+Thanks for your suggestion. In this study, texture profile analysis was investigated by a Texture Analyzer (TA.XT PLUS/50, STABLEMICVO, UK) using a P/0.5R cylindrical probe, and corresponding description has been added to the manuscript in Line 526 marked in red font.	2	1	What equipment was used for the texture profile analysis?	1	2	gels8040212_makarova	1
+The gel without the emulsion was taken as the emulsion blank gel, which was the gel formed only by gel continuous phase.	2	1	Describe the emulsion blank gel.	1	2	gels8040212_makarova	1
+Wet gel were the emulsion gels we obtained, which had a three-dimensional network structure that can hold a large amount of water. The corresponding Line 570 in the manuscript has been revised to a more accurate expression, emulsion gels. Thanks again for your careful review.	2	1	"What does the term ""wet gel"" mean?"	1	2	gels8040212_makarova	1
+Thanks for carefully checking. Whey protein isolate (WPI) has been added in Line 60-61 in the manuscript.	2	1	Line 60: define WPI at first use.	1	2	gels8040212_makarova	1
+Thanks for your constructive comment. Line 84-88 has been removed and it made the manuscript more logically. Thank you again for the good comment.	2	1	Line 84-88: This is a summary of what was carried out and not a discussion, please remove.	1	2	gels8040212_makarova	1
+The emulsions of different droplet size was prepared at WPI concentration of 1.5 wt% and this information has been added in Line 102. This concentration was selected as a suitable emulsifier concentration based on the results in Figure 1a and 1b, and more details can be found in the manuscript.	2	1	Figure 2: Increase the font of the legend.	1	2	gels8040212_makarova	1
+The manuscript has been revised accordingly in Line 159-161.	2	1	Line 166: English check.	1	2	gels8040212_makarova	1
+Figure 3 has been revised accordingly.	2	1	Figure 3: rename using latin alphabet	1	2	gels8040212_makarova	1
+Figure 4 has been adjusted to make the legend clear. In fact, B1, B2 and B3 were in a relatively homogeneous state, and no phase separation had occurred. The shadow in the middle part of the apparent image of the heat-induced gels was because the texture test was carried out before shooting. Unlike TGase-induced emulsion gel had very good elastic texture, the structure of heat-induced gels did not fully recover to its original state due to its soft and less elastic texture after compression.	2	1	Figure 4: Increase legend as it is not visible. B1, B2, and B3 looks biphasic, was this the case?	1	2	gels8040212_makarova	1
+Thanks for your constructive suggestion. Figure 7B has been revised accordingly, that is, a graph was added which displayed the apparent viscosity of all emulsion gels at a fixed shear rate of 0.25/s, showing the differences between groups very visually. Thanks again for your suggestion.	2	1	Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate.	1	2	gels8040212_makarova	1
+In order to choose appropriate emulsifier, we prepare different WPI concentration of 1.0-2.0 wt% and found the emulsion formulated with 1.5 wt% WPI had smaller size and medimum zeta potential, so it was described to the optimize condition. For clarity, we have also revised the corresponding positions in the manuscript.	2	1	Line 474: What is meant by optimize condition?	1	2	gels8040212_makarova	1
+Thanks for your good suggestion. The gel continuous phase was the WPI-gelatin mixture. To more clearly describe the composition of the emulsion gel, Table 3 has been added to the manuscript. The pre-gel mixture, refered to the mixture before gelation occurred, was composed of WPI-gelatin mixture and emulsions, but due to the large number of samples prepared by the single factor experiment, only the general formulation of the emulsion gel was listed in Table 3. And the relevant statements in 4.2.2 and 4.2.3 have been revised in the manuscript.	2	1	Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing? This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.	1	2	gels8040212_makarova	1
+Thanks for your check. It was revised in the manuscript.	2	1	Line 496: remove respectively	1	2	gels8040212_makarova	1
+Thanks for your carefully review. The characterization of emulsions was investigated by the droplet size, polydispersity index (PDI) and zeta potential. This expression has been revised in the manuscript. Point 14: Line 500: Define PDI at first use	2	1	Line 499. Characteristics or characterization?	1	2	gels8040212_makarova	1
+PDI was the polydispersity index and the manuscript has been revised in Line 501.	2	1	Line 500: Define PDI at first use Line 507: Clearance height or gap`?	1	2	gels8040212_makarova	1
+The gap was set to 1 mm. Thank you again for your careful review.	2	1	Line 507: Clearance height or gap`?	1	2	gels8040212_makarova	1
+Thanks for your question. 1% strain was within the linear viscoelastic region in our pre-experiment so it was chosen for other tests.	2	1	Line 512: Was 1% found to be within the viscoelastic region?	1	2	gels8040212_makarova	1
+Thanks for your comment. The statement in 4.2.5 has been revised in the manuscript.	2	1	Line 516: rephase	1	2	gels8040212_makarova	1
+Thanks for your carefully check. Line 533 has been modified which corresponding to the line of revised manuscript.	2	1	Line 530: English check	1	2	gels8040212_makarova	1
+Thanks for your suggestion. In this study, texture profile analysis was investigated by a Texture Analyzer (TA.XT PLUS/50, STABLEMICVO, UK) using a P/0.5R cylindrical probe, and corresponding description has been added to the manuscript in Line 526 marked in red font.	2	1	What equipment was used for the texture profile analysis?	1	2	gels8040212_perova	1
+The gel without the emulsion was taken as the emulsion blank gel, which was the gel formed only by gel continuous phase.	2	1	Describe the emulsion blank gel.	1	2	gels8040212_perova	1
+Wet gel were the emulsion gels we obtained, which had a three-dimensional network structure that can hold a large amount of water. The corresponding Line 570 in the manuscript has been revised to a more accurate expression, emulsion gels.	2	1	"What does the term ""wet gel"" mean?"	1	2	gels8040212_perova	1
+It is our great pleasure to get your professional comments. Thanks very much for your contribution to our manuscript. We take the concerns seriously and comments have been carefully addressed.	2	1	One main request would be to make it clearer what was the formulations and conditions tested.	1	2	gels8040212_perova	1
+Thanks for carefully checking. Whey protein isolate (WPI) has been added in Line 60-61 in the manuscript.	2	1	Line 60: define WPI at first use.	1	2	gels8040212_perova	1
+Thanks for your constructive comment. Line 84-88 has been removed and it made the manuscript more logically. Thank you again for the good comment.	2	1	Line 84-88: This is a summary of what was carried out and not a discussion, please remove.	1	2	gels8040212_perova	1
+The emulsions of different droplet size was prepared at WPI concentration of 1.5 wt% and this information has been added in Line 102. This concentration was selected as a suitable emulsifier concentration based on the results in Figure 1a and 1b, and more details can be found in the manuscript.	2	1	Figure 1: Include in Fig.1c what was the the % of WPI used.	1	2	gels8040212_perova	1
+Thanks for your good comment. Figure 2 has been modified accordingly to make the legend clearer.	2	1	Figure 2: Increase the font of the legend.	1	2	gels8040212_perova	1
+The manuscript has been revised accordingly in Line 159-161.	2	1	Line 166: English check.	1	2	gels8040212_perova	1
+Figure 3 has been revised accordingly.	2	1	Figure 3: rename using latin alphabet	1	2	gels8040212_perova	1
+Figure 4 has been adjusted to make the legend clear. In fact, B1, B2 and B3 were in a relatively homogeneous state, and no phase separation had occurred. The shadow in the middle part of the apparent image of the heat-induced gels was because the texture test was carried out before shooting. Unlike TGase-induced emulsion gel had very good elastic texture, the structure of heat-induced gels did not fully recover to its original state due to its soft and less elastic texture after compression.	2	1	Figure 4: Increase legend as it is not visible. B1, B2, and B3 looks biphasic, was this the case?	1	2	gels8040212_perova	1
+Thanks for your constructive suggestion. Figure 7B has been revised accordingly, that is, a graph was added which displayed the apparent viscosity of all emulsion gels at a fixed shear rate of 0.25/s, showing the differences between groups very visually. Thanks again for your suggestion.	2	1	Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate.	1	2	gels8040212_perova	1
+In order to choose appropriate emulsifier, we prepare different WPI concentration of 1.0-2.0 wt% and found the emulsion formulated with 1.5 wt% WPI had smaller size and medimum zeta potential, so it was described to the optimize condition. For clarity, we have also revised the corresponding positions in the manuscript.	2	1	Line 474: What is meant by optimize condition?	1	2	gels8040212_perova	1
+Thanks for your good suggestion. The gel continuous phase was the WPI-gelatin mixture. To more clearly describe the composition of the emulsion gel, Table 3 has been added to the manuscript. The pre-gel mixture, refered to the mixture before gelation occurred, was composed of WPI-gelatin mixture and emulsions, but due to the large number of samples prepared by the single factor experiment, only the general formulation of the emulsion gel was listed in Table 3. And the relevant statements in 4.2.2 and 4.2.3 have been revised in the manuscript.	2	1	Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing? This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.	1	2	gels8040212_perova	1
+Thanks for your check. It was revised in the manuscript.	2	1	Line 496: remove respectively	1	2	gels8040212_perova	1
+Thanks for your carefully review. The characterization of emulsions was investigated by the droplet size, polydispersity index (PDI) and zeta potential. This expression has been revised in the manuscript.	2	1	Line 499. Characteristics or characterization?	1	2	gels8040212_perova	1
+PDI was the polydispersity index and the manuscript has been revised in Line 501.	2	1	Line 500: Define PDI at first use	1	2	gels8040212_perova	1
+The gap was set to 1 mm. Thank you again for your careful review.	2	1	Line 507: Clearance height or gap`?	1	2	gels8040212_perova	1
+Thanks for your question. 1% strain was within the linear viscoelastic region in our pre-experiment so it was chosen for other tests.	2	1	Line 512: Was 1% found to be within the viscoelastic region?	1	2	gels8040212_perova	1
+Thanks for your comment. The statement in 4.2.5 has been revised in the manuscript.	2	1	Line 516: rephase	1	2	gels8040212_perova	1
+Thanks for your carefully check. Line 533 has been modified which corresponding to the line of revised manuscript.	2	1	Line 530: English check	1	2	gels8040212_perova	1
+Thanks for your good suggestion and we have improved it.	2	1	The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424. Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier. To have a meaningful comparison, the authors need to show the expression level of these two genes in the drought assay (Lines 400-417). As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing. In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal. This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.	1	2	genes13030404_makarova	1
+Thanks for your good suggestion and we have added the possible reason in the discussion.	2	1	It is interesting to see that only MDA content increases in response to drought. The possible mechanism/reason should be included in the discussion.	1	2	genes13030404_makarova	1
+Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	The discussion needs to be further expanded. Esp. RNA-Seq results need to be further discussed.	1	2	genes13030404_makarova	1
+Thanks for your good suggestions on this point. In our revised manuscript, we have made corresponding modifications to summary our work.	2	1	There should be a summary (concluding statement at the end). This manuscript ends very abruptly.	1	2	genes13030404_makarova	1
+Thanks for your good suggestion. We removed “and so on” in our revised manuscript.	2	1	"Pg 1 line 45: the wording ""and so on"" is awkward and imprecise. Please rephrase and indicate what other processes are being referred to."	1	2	genes13030404_makarova	1
+Thanks for your reminding. We modified “As we all know” into “More and more research reveals that”.	2	1	Pg 2 line 67: don't assume that everyone reading the manuscript are aware of miRNA roles in plant stress physiology.	1	2	genes13030404_makarova	1
+It is really true as you mention that the details of methods is needed. But considering that this method is also expressed in thus way in our previous articles (Ren L, et al (2021) Genome-Wide Identification of TCP Transcription Factors Family in Sweet Potato Reveals Significant Roles of miR319-Targeted TCPs in Leaf Anatomical Morphology. Front. Plant Sci. 12: 686698. doi: 10.3389/fpls.2021.686698), we just cite the original source here. Point 4: Pg 6 line 239: I don't understand the point being made here.	2	1	Pg 3 Plasmid and sweet potato genetic transformation section: Include relevant protocols instead of just referring to another manuscript. Citing the original source is required but so are details needed to reproduce and evaluate the soundness of the manuscript.	1	2	genes13030404_makarova	1
+Thanks for your question Target mimicry is a common method to study the function of miRNA. There is a non-coding protein gene IPS in Arabidopsis, and its transcription RNA can specifically bind to miRNA through sequence complementarity, but it forms incomplete complementary bubble structure at the cleavage site of miRNA, that is, miRNA cannot cleavage IPS RNA, thus inhibiting the degradation of miRNA to its target gene mRNA. IPS gene is equivalent to the target mimicry. (Franco-Zorrilla et al., 2007) 2 1	2	1	"Pg 6 line 239: I don't understand the point being made here. ""sequester the normal expression"" is nonsensical. Please clarify and rephrase."	1	2	genes13030404_makarova	1
+Thanks for your good suggestion and we have improved it.	2	1	The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424. Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier. To have a meaningful comparison, the authors need to show the expression level of these two genes in the drought assay (Lines 400-417). As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing. In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal. This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.	1	2	genes13030404_perova	1
+Thanks for your good suggestion and we have added the possible reason in the discussion.	2	1	It is interesting to see that only MDA content increases in response to drought. The possible mechanism/reason should be included in the discussion.	1	2	genes13030404_perova	1
+Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	The discussion needs to be further expanded. Esp. RNA-Seq results need to be further discussed.	1	2	genes13030404_perova	1
+Thanks for your good suggestions on this point. In our revised manuscript, we have made corresponding modifications to summary our work.	2	1	There should be a summary (concluding statement at the end). This manuscript ends very abruptly.	1	2	genes13030404_perova	1
+Thanks for your good suggestion. We removed “and so on” in our revised manuscript.	2	1	"Pg 1 line 45: the wording ""and so on"" is awkward and imprecise. Please rephrase and indicate what other processes are being referred to."	1	2	genes13030404_perova	1
+Thanks for your reminding. We modified “As we all know” into “More and more research reveals that”.	2	1	Pg 2 line 67: don't assume that everyone reading the manuscript are aware of miRNA roles in plant stress physiology.	1	2	genes13030404_perova	1
+It is really true as you mention that the details of methods is needed. But considering that this method is also expressed in thus way in our previous articles (Ren L, et al (2021) Genome-Wide Identification of TCP Transcription Factors Family in Sweet Potato Reveals Significant Roles of miR319-Targeted TCPs in Leaf Anatomical Morphology. Front. Plant Sci. 12: 686698. doi: 10.3389/fpls.2021.686698), we just cite the original source here.	2	1	Pg 3 Plasmid and sweet potato genetic transformation section: Include relevant protocols instead of just referring to another manuscript. Citing the original source is required but so are details needed to reproduce and evaluate the soundness of the manuscript. The same applies to other parts of the materials and methods:	1	2	genes13030404_perova	1
+Thanks for your question Target mimicry is a common method to study the function of miRNA. There is a non-coding protein gene IPS in Arabidopsis, and its transcription RNA can specifically bind to miRNA through sequence complementarity, but it forms incomplete complementary bubble structure at the cleavage site of miRNA, that is, miRNA cannot cleavage IPS RNA, thus inhibiting the degradation of miRNA to its target gene mRNA. IPS gene is equivalent to the target mimicry. (Franco-Zorrilla et al., 2007) 2 1	2	1	"Pg 6 line 239: I don't understand the point being made here. ""sequester the normal expression"" is nonsensical. Please clarify and rephrase."	1	2	genes13030404_perova	1
+Comments #1 (gene expression assay) and #3 (proper discussion on RNA-Seq data) aren't properly addressed.	3	2	Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	genes13030404_perova	1
+The colour refer to the SEEBASE depth to basement, the legend has been added • Figure 2: The left spine (or border) does not appear on the pdf reviewed.	2	1	Figures. Figure 1: Are colors in Figure 1 A related to the hydrostratigraphy or depicting topography? If the latter applies, make sure to provide a scale.	1	2	geosciences12010037_makarova	1
+Thanks, the figure have been entirely revisited and divided into four subfigures.	2	1	Figure 2: The left spine (or border) does not appear on the pdf reviewed. This figure could be improved by adding shaded satellite image to help the reader compare the mapped drainage systems (which were automatically mapped) and fault traces/lineaments with the real features observed.	1	2	geosciences12010037_makarova	1
+The figure has been redrawn and the magnetic lineaments indicated • Figure 4: This figure could go in the appendix.	2	1	Figure 3: The text (lines 258-259) mentions magnetic lineaments as prominent features, which can be caused by faulting. Such features are hard to visualize. It would certainly help if they were highlighted directly on the figure.	1	2	geosciences12010037_makarova	1
+Thank you. We hesitated on this point but decided to keep the figure in as the quality of the seismic in the shallow level is really crucial to the discussion. We need to know if the interpretation is reliable, especially in this shallow zone that is not well imaged by this type of data. A misinterpretation can led to the targeting of circulation pathways that are in fact only artefacts.	2	1	Figure 4: This figure could go in the appendix. It is good to let the reader know that not all seismic surveys provided the same quality, but this is more a technical point.	1	2	geosciences12010037_makarova	1
+We removed the seismic interpretation of figure 7G (cross) that were misleading the reader and add the interpretation of the fluid leakage features on the images	2	1	Figure 7: What is special about the locations highlighted by the markers (crosses on Figure 7B and square markers on Figure 7C). At this scale, the reader can’t see much detail.	1	2	geosciences12010037_makarova	1
+this is due to the different seismic acquisition technics and processing applied in each survey as well as the fact that our interval of interest is shallow and not fitting with the interval of interest the companies tried to best image with those datasets”	2	1	Lines 271-272: Why is the signal quality so variable, and how were the categories (excellent, average and poor) defined?	1	2	geosciences12010037_makarova	1
+We added “as the presence of fluid or gas can locally alter the seismic signal”	2	1	Lines 323-324: The authors should explain why low magnitude local seismic signals can be believed to represent permeable pathways promoting water or gas migration.	1	2	geosciences12010037_makarova	1
+We added: “We define as recent tectonic activity the events that are recorded at the surface and can alter the surface with active tectonic features, such as modifications of the drainage system” • Discussion: One general comment, could the use of other tracers (radon, of stable isotopes of water) yield also insight on the origin of the fluids sampled in this study?	2	1	Line 360: Here and throughout the text, the word ‘recent’ is used to describe some tectonic activity. It would be good define what recent means in the context of this study. An indication is provided in the Discussion (Line 414), but it would go to state this value upfront in the text.	1	2	geosciences12010037_makarova	1
+Radon cannot give this information since with the short half-live of 3.8days it's signal comes only from the direct vicinity of the sampled location. Stable isotopes of water could in principle do this if there is a geothermal signature on them (such as plotting far left to the meteoric water line - see inset in Figure 1 of McIntosh and Ferguson, 2020 – Geophysical research Letter) - but we have not seen such a signature. We may mention that in the text - but for such a signature to be discernible the bulk of the water needs to be geothermal, and what we see is only a small admixture.	2	1	Discussion: One general comment, could the use of other tracers (radon, of stable isotopes of water) yield also insight on the origin of the fluids sampled in this study?	1	2	geosciences12010037_makarova	1
+The revisited figure 43 is now providing this information Seismic reflection 2D lines (1989-2015).	2	1	Aeromagnetic survey data acquired over the last 50+ years were employed for mapping structural and lithological features. Although the aeromagnetic data is said to be employed for structural interpretation, details somewhat sketchy. Reference is made to “…prominent […] magnetic lineaments which can be caused by faulting”, and “…several circular low magnetization anomalies, approximately two kilometers in diameter, aligned with northwest trending lineaments”. It would be nice to include a map with these features and lineaments interpreted from the aeromagnetic datasets. This would also tie the observations based on this dataset closer to the seismic interpretation (see below).	1	2	geosciences12010037_makarova	1
+Thank you, the details comments on those figures have been followed and new figures are provided Helium measurements from groundwater samples.	2	1	Seismic reflection 2D lines (1989-2015). 8500 km. Interpretation of shallow horizons and faults using all publicly available geological and geophysical data of the Beetaloo region. Fault polygons defined for five horizons. Depth conversion using check-shot velocities from 26 wells. See comments to Figure 4. The depth maps provided in Figure 5 should be supplemented by isochore maps for the rock volumes between pairs of mapped reflectors. See also comments to Figure 7.	1	2	geosciences12010037_makarova	1
+"There is no other reference to date to the ""primary documentation"" but the link to the data (they should now be online) which are in the fact sheets."	2	1	Helium measurements from groundwater samples. With respect to the helium measurements, no detailed documentation of where these samples were collected is provided beyond referring to Geological and Bioregional Assessment Program (2021a) Fact sheet 12, and Geological and Bioregional Assessment Program (2021b) Regional tracer results from the Cambrian Limestone Aquifer. These are both summary documents. Please provide a reference to the primary documentation.	1	2	geosciences12010037_makarova	1
+We have redone the figure 2 and added explanations on the process (see reply to comments on figure 2). Figure 8 should now be easier to read The surface drainage network line dataset was indeed based on the simplified (n=130) network following the following approach: Small segments with less than 10 nodes were automatically removed in SKUA (Paradigm/Emerson Trademark) and one iteration of smoothing was applied that further removed small irregularities without changing the overall orientation of major drainage lines.	2	1	Fault and surface drainage network mapping (tectonic geomorphology). The drainage network was extracted automatically. It is not clear if the “simplified” drainage network shown in Figure 2 was also extracted automatically. The data is summarized in Figure 8 (see also comments to Figure 8 below). It is not clear if the surface drainage network line dataset is based on the “unfiltered” or “simplified” data shown in Figure 2. Since n=130, one can assume the latter. It is however not made clear how this “simplified” network was generated, but there appear to be a substantial number of instances where the “simplification” provides apparent mismatches with the original data (see examples included in comments to Figure 2 below). The accuracy of the resulting plot can therefore be questioned. If the “simplified” network was generated automatically, I encourage the authors to do a manual QC of the results.	1	2	geosciences12010037_makarova	1
+There is indeed an error on the figure referencing, we modified to Figure 3B where the lineaments are now clearly indicated.	2	1	Line 303: “We reviewed the seismic data in detail at the intersections with north-northwest trending lineaments observed on the magnetic dataset (Figure 4).” No such lineaments are shown in Figure 4.	1	2	geosciences12010037_makarova	1
+We indeed need to add this link- we will put a link in place upon acceptance of the paper Figures Figure 1A.	2	1	Line 582. Link for supplementary materials does not work.	1	2	geosciences12010037_makarova	1
+done, we also modified the fault legend to add clarity - Replace white labels with black lettering with no labels and white lettering.	2	1	- Poor visual discretization of main surface fault traces and Post Wilton geophysical faults (maybe use different colour for the two?)	1	2	geosciences12010037_makarova	1
+We separated the legends from figures 1A and 1B as to add clarity. This article is focused on the Beetaloo subbasin and we use the direct input for Geoscience Australia in this stratigraphic column. We are not willing to add complexity by presenting in detail the Basins that are covering the Beetaloo Sub-Basin. We removed from the legend the details on the different basins as we understand that it is leading the reviewer in directions we are not exploring on the CLA stratigraphy- we do not refer this complexity in the results and discussion.	2	1	Figure 1B - Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin. Although this is described in the caption, it is not easy to grasp for someone not familiar with the stratigraphy in the area without spending time some time with paper and pencil.	1	2	geosciences12010037_makarova	1
+Quality improved - Very, very overloaded figure, screen-dump from a mapping programme?	2	1	Figure 2 - Poor quality figure (low resolution) in the version available to the reviewer.	1	2	geosciences12010037_makarova	1
+Thanks, the figure have been entirely revisited and divided into four subfigures as advised below -Consider splitting into several maps, e.g.	2	1	- Very, very overloaded figure, screen-dump from a mapping programme?	1	2	geosciences12010037_makarova	1
+This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control. In this process, minor mismatches of the automated simplified lines were removed and small drainage segments were connected along major lineaments for further simplification and reduction of the dataset. The adopted approach was: Small segments with less than 10 nodes were automatically removed in SKUA (Paradigm/Emerson Trademark) and one iteration of smoothing was applied that further removed small irregularities without changing the overall orientation of major drainage lines. We added this description of the adopted approach in the legend Figure 3 -Very small/illegible typeface on map coordinates – consider simplifying scales and use larger typeface.	2	1	- How the authors arrive at the simplified drainage network is not very clear	1	2	geosciences12010037_makarova	1
+figure redrawn - Replace white labels with black typeface with white typeface.	2	1	Figure 4 - Poor quality figure (low resolution) in the version available to the reviewer.	1	2	geosciences12010037_makarova	1
+Well label typos modified -Thin grey seismic lines are not explained in the legend or caption.	2	1	-Many well-name labels are hard difficult to read/illegible. Consider replacing well names on the map with numbers at the well position (white circles, black typeface) and add a table in the legend listing number and corresponding name of the well.	1	2	geosciences12010037_makarova	1
+Legend added for those lines -The outline of the Eastern and Western Beetaloo Basin in Figure 3 and 4 should be kept identical in order to position Figure 6 A, B and C in relation to the seismic lines shown in Figure 4.	2	1	-Thin grey seismic lines are not explained in the legend or caption. Why are these not classified in terms of signal quality?	1	2	geosciences12010037_makarova	1
+sochores added as supplementary material	2	1	Consider adding isochore maps for intervals between interpreted reflectors. These often highlight tectonic accommodation space creation better than maps.	1	2	geosciences12010037_makarova	1
+done, legend cleared from information not represented	2	1	d) Explain/improve labels (what do you mean by “ntspr_2M_gw”? “Coast_10million”? “frameworkboundaries”? “State_Borders_10million”. Amend labels or explain in caption.	1	2	geosciences12010037_makarova	1
+checked and clarified Clean up map: 1.	2	1	e) Several items in the legend appear to have the same or very similar signatures.	1	2	geosciences12010037_makarova	1
+Clearly identified in the legend and representation 2.	2	1	Clean up map: Not possible to differentiate post-Wilton fault types properly in the figure.	1	2	geosciences12010037_makarova	1
+removed	2	1	Could not find “Coast_10million”, “frameworkboundaries” or “State_Borders_10million” on the map. If these items are present, please use a more contrasting signatures to make it more visible. If they are not present on the map, remove these items from the legend.	1	2	geosciences12010037_makarova	1
+Outline size modified and homogenised with other figures 4.	2	1	Consider replacing the purple outline of the eastern and western sub-basin with light grey shading.	1	2	geosciences12010037_makarova	1
+Red selected 5.	2	1	Consider using the most prominent colour (red) for the possible fluid escape features as this is the key element of the map. Bright green might also bee an option.	1	2	geosciences12010037_makarova	1
+modified	2	1	Not possible to differentiate “ntspr_2M_gw” (whatever that is) and “PossibleFluidEscapeStructures” (same colour).	1	2	geosciences12010037_makarova	1
+modified accordingly Consider adding a simple conceptual sketch highlighting the stratigraphic position of the different faults (and possible fluid escape structures– this would also help visualizing which stratigraphic intervals the potential fluid escapes originate from and which stratigraphic intervals are potentially connected along potential fault-related fluid flow pathways.	2	1	The link between interpreted faults and fluid or gas escape features is a key feature of this figure. The map differentiates between “BaseCambrian seismic faults”, Post-Wilton “Strike-slip” and “Reverse” faults, and “FAULTS”. The categorization is not adequately explained (e.g. what differentiates “FAULT” from a “BaseCambrianSeismic fault”?).	1	2	geosciences12010037_makarova	1
+With the new classification the reader should be able to better read the paper and can now directly refer to the stratigraphic table Figure 8.	2	1	Consider adding a simple conceptual sketch highlighting the stratigraphic position of the different faults (and possible fluid escape structures– this would also help visualizing which stratigraphic intervals the potential fluid escapes originate from and which stratigraphic intervals are potentially connected along potential fault-related fluid flow pathways.	1	2	geosciences12010037_makarova	1
+Done, figure clarified as well Figure 9.	2	1	Please provide a more informative caption to what these plots show. What is included in the “Drainage and structural lineament analysis” plot?	1	2	geosciences12010037_makarova	1
+done, we also totally revisited this figure as to have a cleared output	2	1	Figure 9. Poor resolution in the copy provided for the review. Consider using higher contrast colours or thicker lines to highlight different faults on the map.	1	2	geosciences12010037_makarova	1
+The helium measurements have been added as supplementary data and the author made a clear difference between this study Helium measurements and other tracer studies we are citing.	4	1	With respect to the helium measurements, the references to the primary documentation requested by the reviewer in comments to v.1 of the manuscript are not provided, nor do the authors provide any comment as to why they deem this unnecessary. Section 4.3 Helium concentrations and isotopic composition measured in groundwater, is framed around results from previous studies. Providing references to these original studies (including access to the actual data that allow objective verification) rather than referencing to summary papers would be more in line with scientific publishing practice. If the primary documentation is not accessible, the authors should state this clearly.	3	2	geosciences12010037_makarova	1
+Done, thank you	4	1	Figure 4. Seismic reflection data quality is ranked and colour-coded as “Excellent”, “Average” and “Poor”. Where does the fourth category, “Fair”, which also is a qualitative category, fit in, and why not include this category in the legend?	3	2	geosciences12010037_makarova	1
+The isochores have been added in the seismic supplementary dataset	4	1	Figure 5. Isochron maps have been included in the supplementary material, the initial review suggested isoCHORE maps. Including these is maybe not critical, but as mentioned in the initial review, thickness maps sometimes highlight structural lineaments quite well. I don't know if this is the case here, but if they do, including isochore maps would enhance the readers' understanding of basin geometry and -infill.	3	2	geosciences12010037_makarova	1
+Post Wilton “Strike-slip” and “Reverse” faults have been redrawn in all figures (Figure 1, 7, 9) with a new colour code, the fluid escape features are now shown as red dots in the legend as well.	4	1	If I understand the map correctly, fluid escape features on the map are shown as red dots. Why does the legend show these as thick red lines?	3	2	geosciences12010037_makarova	1
+Number of measurements added in each plot. Figure 2a correspond to the SEEBASE structural lineaments (n=681).	4	1	Figure 8. Please keep the number of measurements included in each plot. The plot “Surface drainage network from this study” is identical to the original figure and should have n=130 measurements. According to the new caption shows the unfiltered surface drainage network. To my understanding this should correspond to Figure 2 a, which shows substantially more than 130 lineaments. How did you pick the ones you measured? Please clarify.	3	2	geosciences12010037_makarova	1
+we are very sorry for the mistake, and the manuscript was modified accordingly.	2	1	In the Abstract you state that “two patients during their course of Rivaroxaban presented severe hematoma”, but in Table 1 it appears that the two hematomas occurred in the enoxaparin group? Please clarify.	1	2	healthcare10030476_makarova	1
+The criticism of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	The introduction is not well written. The structure is unclear, and the authors repeat themselves many times. For example, the section begins with introducing inherited thrombophilias and VTE risk in these conditions, then proceeds to surgery, then back again to thrombophilia where it repeats itself and even states other numbers for prevalences and VTE risks than previously stated. Furthermore, suitable references are lacking in some places. For the present study to be acceptable for publication, the introduction should be thoroughly revised.	1	2	healthcare10030476_makarova	1
+The observation of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	The Discussion section could benefit from a language revision, especially in the latter part, to make the flow better. Also, a brief discussion on the limitations of your study should be included.	1	2	healthcare10030476_makarova	1
+The observation of the reviewer has been accepted and the table 1 was modified accordingly.	4	1	"It is still not clear how thrombophilia was defined. You write (page 3, lines 129-130): ""Only women with thrombophilia defined as a factor correlated with thrombotic phenomena linked to congenital or acquired factors. [21,40-43]"" I suggest you alter the sentence, e.g. to something like: ""Thrombophilia was defined as either the diagnosis of an inherited thrombophilia, a personal history of VTE or a familial history of VTE."" If that is how you defined thrombophilia?"	3	2	healthcare10030476_makarova	1
+The new manuscript has been evaluated by an expert of English language.	4	1	Also, another round of language editing may be suitable.	3	2	healthcare10030476_makarova	1
+The observation of the reviewer has been accepted and the table 1 was modified accordingly.	4	1	"I have one major comment. It is still not clear how thrombophilia was defined. You write (page 3, lines 129-130): ""Only women with thrombophilia defined as a factor correlated with thrombotic phenomena linked to congenital or acquired factors. [21,40-43]"" I suggest you alter the sentence, e.g. to something like: ""Thrombophilia was defined as either the diagnosis of an inherited thrombophilia, a personal history of VTE or a familial history of VTE."" If that is how you defined thrombophilia?"	3	2	healthcare10030476_perova	1
+The new manuscript has been evaluated by an expert of English language.	4	1	Also, another round of language editing may be suitable.	3	2	healthcare10030476_perova	1
+we are very sorry for the mistake, and the manuscript was modified accordingly.	2	1	In the Abstract you state that “two patients during their course of Rivaroxaban presented severe hematoma”, but in Table 1 it appears that the two hematomas occurred in the enoxaparin group? Please clarify.	1	2	healthcare10030476_perova	1
+The criticism of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	The introduction is not well written. The structure is unclear, and the authors repeat themselves many times. For example, the section begins with introducing inherited thrombophilias and VTE risk in these conditions, then proceeds to surgery, then back again to thrombophilia where it repeats itself and even states other numbers for prevalences and VTE risks than previously stated. Furthermore, suitable references are lacking in some places. For the present study to be acceptable for publication, the introduction should be thoroughly revised.	1	2	healthcare10030476_perova	1
+The observation of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	The Discussion section could benefit from a language revision, especially in the latter part, to make the flow better. Also, a brief discussion on the limitations of your study should be included.	1	2	healthcare10030476_perova	1
+The quoted literature has been adapted to the ACS Style Guide. For that, the authors used to software package Zotero.	2	1	The citations are not formatted according to the journal guidelines. Please revise.	1	2	healthcare10040723_perova	1
+STD are included in the all tables were indicated: sum+STD has been added in the legend.	2	1	Include the full form of STD described in the results.	1	2	healthcare10040723_perova	1
+In Table 3 (=) has been changed into (0). It was a typo, sorry.	2	1	Table 3 “Health apps and computer algorithms are for patients disturbing (=)…” should “(=)” be “(0)”?	1	2	healthcare10040723_perova	1
+Line 252: 7/24 has been changed into 24/7.	2	1	Line 252, 24/7 is the commonly accepted form of 7/24.	1	2	healthcare10040723_perova	1
+Line 267 and 268, the description of figure 1 “focused on senses” means that the diagnostic workup should be more focused on the physical examination (the doctor’s senses) rather than based on technical diagnostics. In the revised manuscript has been added in line 270 “and use their senses in physical examinations rather than rely on impersonal technical tools for the diagnostic work-up”.	2	1	Line 267 and 268, the description of figure 1 is not clear. Does “focused on senses” mean in-person patient-doctor interaction?	1	2	healthcare10040723_perova	1
+In Line 266/267 the term “undermine” was wrong. It has been changed into “ease”. Thank you for the hint.	2	1	Table 4 data suggest Digital networks increase doctor nurse communication but Line 266 says it undermines the same. Can you clarify?	1	2	healthcare10040723_perova	1
+Table 5: The students could rank their answer. 1 = don’t know, 2= false up to 7 = fully agree. The statistical calculation was done only for ranks 2 to 7, excluding the students, who could not or didn’t want to answer the question. The authors knew from the qualitative study part, that some students had no knowledge about AI. Therefor the “don’t know” option was included. In the first draft of the manuscript the legends contained a mistake, because the “fully agree” option had the highest number (7) but not 3. This has been changed in the revised manuscript.	2	1	Table 5, what does numbers 1 through 7 represent?	1	2	healthcare10040723_perova	1
+2. The reviewer discussed in much detail which design might better describe the method used in our study. In the sequential exploratory strategy a qualitative component is followed by a quantitative component (Creswell, JW. ; Plano Clark, VL. Designing and Conducting Mixed Methods Research. Thousand Oaks, CA: Sage; 2011). Aspects of the organizational environment (e.g., creative problem solving) were identified in the qualitative component and statistically confirmed or rejected in the preceding quantitative study phase, which is exactly what the authors did in this study. However, the reviewer suggested that the convergent parallel design might fit our methodology even better. In the convergent parallel strategy, quantitative and qualitative data collection occurred concurrently. The components are given equal weight, and the two datasets are analyzed and compared in parallel. Quantitative data identified factors statistically, which were integrated with qualitative data from in depth interviews with a subset of participants to understand how and why the identified factors influenced participants’ attitudes. In the method’s section of our study we lined out that the results of the first qualitative study part were the basis for the development of the online survey. Based on the qualitative study part, the questions for the qualitative study were formulated when students’ attitudes were expressed frequently. In this sense, the first (quantitative) phase informs the next, but the reviewer is right, that both phases are interlinked. Although our study integrates components of both strategies, the authors still think, that the sequential exploratory strategy describes our methods better than the convergent parallel design (see also: Curry et al Circ. Cardiovasc, Qual. Outcomes 2013, 6:119-123). As suggested by the reviewer, the authors specified in the method section that we used an additive rather than a parallel integrative strategy.	2	1	The fundamental issue that needs clarity is whether or not the study design was ‘exploratory mixed methods design’ or ‘convergent parallel design.’ As far as exploratory mixed methods design is concerned, the researcher begins with qualitative data and then collects quantitative information. Typically, in this design, the researcher presents the study in two phases, with the first phase involving qualitative data collection and the second phase involving quantitative data. In this regard, your study fits with the exploratory nature. In an exploratory sequential mixed-methods design, the researcher emphasizes the qualitative data (QUAL) more than the quantitative data (quan). This emphasis may reveal by presenting the overarching question as an open-ended question or discussing the qualitative results in more detail than the quantitative results. However, in this manuscript, I have not seen elements of these characteristics in the paper. As I have seen a complementary role of the qualitative and quantitative pieces is apparent instead of the qualitative piece informing the quantitative pieces. For example, the description of the study design says the following. Page 2, lines 90-92, ‘This was then integrated into the second study phase consisting of a nationally representative sample of the same sort of cohort.’ This description shows the integration of the two phases instead of the preceding phase informing the next phase. From the explanation in the study design and the presentation of your results, it sounds to me as if the mixed-methods nature of this study fits more with the convergent parallel design. Please, give your readers more information that conveniences the exploratory nature.	1	2	healthcare10040723_perova	1
+The manuscript has been changed accordingly.	2	1	In the abstract, lines 23-27, I suggest to use the following expression or something like that. Around 38% of the students felt ill-prepared and could not answer AI-related questions because digitization in medicine and AI are not a formal part of the medical curriculum.	1	2	healthcare10040723_perova	1
+The authors followed the reviewer’s suggestions and re-wrote this para as follows: “The overall objective of this study was to investigate the today’s medical students attitudes towards AI and other digital working tools. We wanted to understand if age, gender, semester level and curriculum type influences their views. This study also assembled information on students’ understanding of AI algorithms and digital applications in health care and assessed their level of confidence in working alongside these tools after graduation in patient care. It is our belief that this information may possess the means to employ digital tools including AI into the curriculum of medical students efficiently, enhancing their confidence in using them and therefor to better equip our future physicians with sufficient knowledge.” On page 2, line 111, the phrase ‘to affirm or dismiss,’ does not make sense.	2	1	Page 2, line 42, ‘the computation of compounding factors,’ I did not understand what it means. I suggest the authors to replace this with appropriate phrase.	1	2	healthcare10040723_perova	1
+The phrase in question has been replaced by “to confirm”.	2	1	On page 2, line 111, the phrase ‘to affirm or dismiss,’ does not make sense. Replace this phrase with a more appropriate phrase.	1	2	healthcare10040723_perova	1
+The authors choose 4 pre-clinical and 4 clinical students. The intention was - as outlined in the initial manuscript version - “… to understand how they perceive the subject of interest”. The wording has been specified, to clarify that in total 8 students tested the items but not 4 as the reviewer assumed. This number was considered sufficient a) to detect inconsistencies and b) to verify the correct meaning of the questions. AG and JE are two of the authors (first letter of the first and last name). In order to avoid any misunderstanding, this has been changed to “the authors” in the revised manuscript.	2	1	Page 3, lines 117-118, the statement has two issues. ‘The items were then tested through a pilot study consisting of a 117 group of 4 pre- and clinical students, mediated by AG and JE’ First, why did you use only 4 students for pilot testing? Second, what do ‘AG’ and ‘JE’ represent?	1	2	healthcare10040723_perova	1
+Convenience sampling is defined as a method to collect data from a conveniently available pool of respondents. We used it, because it is the most commonly used sampling technique as it's incredibly prompt, uncomplicated, and economical.	2	1	Page 3, line 132, it says, Convenience sampling was used. I would ask, why did you use cconvenience sampling?	1	2	healthcare10040723_perova	1
+The captation has been changed to “Characteristics of the study cohort.	2	1	Page 4, table 1, the caption and the table content do not match. Revise either of them.	1	2	healthcare10040723_perova	1
+The sampling procedure was clearly described in the initial manuscript. For phase 1: “All participants of the first phase were students from from 1st to 6th year (undergraduate 1st to 2th year, graduate 3th to 6th year) from German Universities. The inclusion criteria were to actively study medicine and their agreement for their voluntarily participation. In the same way, the exclusion criteria were to have suspended their studies, as well as other exceptional situations. Prior to start, informed consent was obtained, which was followed by the collection of telephone numbers and email addresses. They were selected purposely and consecutively, in part by snowball until theoretical saturation was reached.” For phase 2: “For the second, quantitative study phase identical inclusion/exclusion criteria applied.” This referred to medical students of all semesters, excluding students from other faculties. “The online survey was sent to all medical faculties in Germany from which most forwarded the survey invitation by email to about 80 000 medical students to fulfill the principle of maximum diversity through convenience sampling method. Each contained an invitation letter and an information sheet.” Surely, sampling procedures in mixed method studies are phase depended different.	2	1	In the methods section, clearly describe the sampling procedure for the quantitative and qualitative study parts separately. Also describe the final study sample for both. I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.	1	2	healthcare10040723_perova	1
+The statement has been changed into: “The focus on German educational system and the fact that only a small fraction of the total number of medical students filled out the online survey makes a generalization of the answers difficult”.	2	1	Page 12, line 389, the statement which says, ‘The non-probabilistic sampling makes a generalization of the answers difficult,’ is confusing. I suggest to re-write this sentence.	1	2	healthcare10040723_perova	1
+The authors agree and the phase “so called” has been removed.	2	1	I suggest to remove the phrase ‘so called,’ that has been used in the conclusion section or any other section.	1	2	healthcare10040723_perova	1
+The authors agree and this statement has been re-written in the revised manuscript version.	2	1	Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement. Re-write this sentence clearly.	1	2	healthcare10040723_perova	1
+The authors fully agree. Therefore, the conclusion paragraph has been split into the “Conclusion” paragraph, in which the study’s conclusions has been outlined, and into a the new para “Implications” where the usefulness of the integration of eHealth aspects (including AI) into the curriculum is discussed.	2	1	I suggest the authors to include a short section including the implications of the study after the conclusions.	1	2	healthcare10040723_perova	1
+We have modified this part in revised manuscript (Line 233-236). This sentence has been changed to “A suspension of A. rhizogenes strain GV3101 containing the pORER4-OfSPL11-GFP recombinant plasmid was aspirated using a 1 mL range syringe (needle not included), and the suspension was gently injected into the abaxial surface of four-week-old tobacco leaves avoiding the leaf veins.” Point 2: Rephrase lines 266-268 is unclear; wording, grammar and punctuation are wrong.	2	1	In line 227 “back of tobacco” is not clear be more specific.	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 38).	2	1	Minor corrections In line 38 replace “Fortunately” with However.	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we have replaced this word in the revised version (Line 63).	2	1	In line 63 replace “trancroptoma” with transcriptome.	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 65)	2	1	In line 65 replace “And” with In addition.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 69).	2	1	In line 69 add period after assay.	1	2	horticulturae8050412_makarova	1
+We had made correction in the manuscript (Line 83-84). The statements of “RNA extractions” were corrected as “total RNA was extracted using the RNA prep Pure Plant Plus Kit (Tiangen, Beijing, China).” Point 10: In line 91 what brand was Taq polymerase from?	2	1	In line 83 what extraction method was used? Mention kit of reagent brand.	1	2	horticulturae8050412_makarova	1
+We had made correction in the manuscript (Line 92). Taq polymerase was purchased from TaKaRa Biotechnology, Inc, Dalian, China.	2	1	In line 91 what brand was Taq polymerase from?	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 102-103). The pORER4-GFP vector and the plant expression vector OfSPL11-GFP were purchased from Tsingke, Biotechnology, Inc, Hangzhou, China.	2	1	In line 100 replace “ligated” with was ligated and quote a reference for pORERA-GFP vector or mention brand if it is commercial.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction carefully (Line 115 and 270).	2	1	In line 113 replace “psoup” with pSOUP and “tobaccos” with tobacco plants.	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 119 and 276).	2	1	In line 117 and 267 replace “vacuuming penetration” with vacuum infiltration.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 129).	2	1	In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 136).	2	1	In line 129 replace “strains” and “strain” with lines and line.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction carefully (Line 138).	2	1	In line 131 delete the period after experiments.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 145).	2	1	In line 138 Seedling should be lowercase.	1	2	horticulturae8050412_makarova	1
+"We had made correction in the manuscript (Line 156-157). The statements of “the MDA assay kit” were corrected as “the MDA assay kit (Jiancheng Bioengineering Institute, Nanjing, China).” Point 20: In line 154 add ""strain"" after receptor and mention kit brand Response 20: Thank you for your comment."	2	1	In line 149 mention the kit brand.	1	2	horticulturae8050412_makarova	1
+"We had made correction in the manuscript (Line 162 and 163). The statements of “the yeast Y187 receptor strain preparation and transformation kit” were corrected as “the yeast Y187 receptor strain preparation and transformation kit (Coolaber, Beijing, China).” Point 21: In line 169 add ""were done"" at the end."	2	1	In line 154 add “strain” after receptor and mention kit brand.	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 177).	2	1	In line 169 add “were done” at the end. A phrase must have a verb.	1	2	horticulturae8050412_makarova	1
+Thank you for your comment, we had made correction in the manuscript (Line 270).	2	1	In line 261 replace “tobaccos” with tobacco plants.	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 271).	2	1	In line 262 replace “were treatment by” with were treated with.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction carefully (Line 273).	2	1	In line 264 replace “indicate” with indicated.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 301).	2	1	In line 292 “The” must be lowercase.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction carefully (Line 327).	2	1	In line 318 “We” must be lowercase.	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 403).	2	1	In line 395 replace “it revealed” with revealed.	1	2	horticulturae8050412_makarova	1
+Thank you for your comment, we had made correction in the manuscript (Line 415).	2	1	In line 406 delete period after stress.	1	2	horticulturae8050412_makarova	1
+We have modified this part in revised manuscript (Line 326-328).	2	1	Line 302: plant leaves instead of plants leaves.	1	2	horticulturae8050412_makarova	1
+genes. We feel sorry for the unclear statement. In our study, we used the amino acid sequence of OfSPL11 to compute the protein model. Secondary structure predictions were conducted on the PRABI (http://www.prabi.fr/spip.php?page=sommaire) protein structure prediction server. The ExPASy (http://web.expasy.org) was used to analyze the molecular weight, theoretical isoelectric point, and other physiochemical properties of the translated protein. Similar methods of operation and efforts can be found in other articles (Islam, 2021). Once again,we sincerely thank you for the recommended method, which we will use in future studies. Islam, M.D.; Rahman, M.M.; Rahman, M.M. ; Jin, X.J. ; Sun, L.L. ; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S. ; Zhang, W.J. ; Sun, D.Z. In Silico and Transcription Analysis of Trehalose-6-phosphate Phosphatase Gene Family of Wheat: Trehalose Synthesis Genes Contribute to Salinity, Drought Stress and Leaf Senescence. 2021, 12, 1652. https://doi.org/10.3390/genes12111652 Point 2: Auhtors report that Actin was used as housekeeping gene.	2	1	Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?	1	2	horticulturae8050412_makarova	1
+We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015). Dheda, K.; Huggett, J.F. ; Bustin, S.A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR. Biotechniques. 2004, 37: 112-119. https://doi.org/10.2144/04371RR03 Czechowski, T.; Stitt, M.; Altmann, T. Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol. 2005, 139: 5-17. https://doi.org/10.1104/pp.105.063743 Hou, H.; Jia, H.; Yan, Q.; Wang, X. Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance. Int. J. Mol. Sci. 2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X. ; Wang, Y.G. ; Bao, Z.Y. ; Zhao, H.B. Identification of Suitable Reference Genes for Gene Expression Normalization in the Quantitative Real-Time PCR Analysis of Sweet Osmanthus (Osmanthus fragrans Lour.). Plos one. 2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.	2	1	Auhtors report that Actin was used as housekeeping gene. Do the authors have performed an efficiency test before using it? Do they tried to use otherHK genes?	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 130).	2	1	Furthermore, I reccomend a text revision since some errors are present in the main text. Line 124: generate instead of generate	1	2	horticulturae8050412_makarova	1
+We have revised the manuscript according to your comments (Line 274-277). This sentence has been changed to “A. tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 3: Rephrase lines 317-318.	2	1	"Line 267-268: Please, check this sentence. It ends with ""then"". Figure 5 caption: activates instead of activate Line 302: plant leaves instead of plants leaves"	1	2	horticulturae8050412_makarova	1
+We had made correction carefully (Line 283).	2	1	Figure 5 caption: activates instead of activate.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 310).	2	1	Line 302: plant leaves instead of plants leaves.	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 319).	2	1	Line 311: plant leaves instead of plants leaves	1	2	horticulturae8050412_makarova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 324).	2	1	"Line 315: maybe a semicolon after ""salt stress"" is appropriate."	1	2	horticulturae8050412_makarova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 366).	2	1	"Line 360: remove the space after ""assay"""	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion. We had made correction carefully (Line 381).	2	1	Figure 9 caption: measure instead of measuring	1	2	horticulturae8050412_makarova	1
+Special thanks to you for your good suggestion, we had made correction in the manuscript (Line 398).	2	1	"Line 392: I suggest to change the sentence in the following way "".. to drive the expression of target"	1	2	horticulturae8050412_makarova	1
+"Thank for your kindly suggestion. We have revised the manuscript according to your comments (Line 456). The statements of “participate in salt stress responses” were corrected as “participate in salt stress.” Point 12: Line 448: check the word ""redycing"" Response 12: Thank you for your comment."	2	1	"Line 446: ""...participate in salt stress""Line 448: check the word ""redycing"" "	1	2	horticulturae8050412_makarova	1
+We have modified this part in revised manuscript (Line 233-236). This sentence has been changed to “A suspension of A. rhizogenes strain GV3101 containing the pORER4-OfSPL11-GFP recombinant plasmid was aspirated using a 1 mL range syringe (needle not included), and the suspension was gently injected into the abaxial surface of four-week-old tobacco leaves avoiding the leaf veins.” Point 2: Rephrase lines 266-268 is unclear; wording, grammar and punctuation are wrong.	2	1	In line 227 “back of tobacco” is not clear be more specific.	1	2	horticulturae8050412_perova	1
+We have modified this part in revised manuscript (Line 326-328). This sentence has been changed to “To better understand why OfSPL11 transgenic lines exhibit enhanced abiotic stress tolerance, we analyzed the expression of known abiotic stress response genes in 3-week-old wild-type and transgenic Arabidopsis.” Point 4: In lines 271-272 and 395-396, how do you explain that salt induced promoter trans-activation?	2	1	Rephrase lines 317-318. Comments as above.	1	2	horticulturae8050412_perova	1
+In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China). The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves. The LUC signal increased profoundly after subjection to HS (37℃), although under normal conditions (room temperature), the LUC signal was very low, which implied that HS activated the promoter activity of LlWRKY39. In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions. Therefore, we suggest that salt stress can induce the activity of the OfSPL11 promoter.	2	1	In lines 271-272 and 395-396, how do you explain that salt induced promoter trans-activation? Is there any published precedent?	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 38).	2	1	Write appropriate brand of kits and reagents, followed by country. Check journal format.	1	2	horticulturae8050412_perova	1
+Thank for your kindly suggestion, we have replaced this word in the revised version (Line 63).	2	1	In line 38 replace “Fortunately” with However.	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 65).	2	1	In line 63 replace “trancroptoma” with transcriptome.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 69).	2	1	In line 65 replace “And” with In addition.	1	2	horticulturae8050412_perova	1
+We had made correction in the manuscript (Line 83-84). The statements of “RNA extractions” were corrected as “total RNA was extracted using the RNA prep Pure Plant Plus Kit (Tiangen, Beijing, China).” Point 10: In line 91 what brand was Taq polymerase from?	2	1	In line 69 add period after assay.	1	2	horticulturae8050412_perova	1
+We had made correction in the manuscript (Line 92). Taq polymerase was purchased from TaKaRa Biotechnology, Inc, Dalian, China.	2	1	In line 83 what extraction method was used? Mention kit of reagent brand.	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 102-103). The pORER4-GFP vector and the plant expression vector OfSPL11-GFP were purchased from Tsingke, Biotechnology, Inc, Hangzhou, China.	2	1	In line 91 what brand was Taq polymerase from?	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction carefully (Line 115 and 270).	2	1	In line 100 replace “ligated” with was ligated and quote a reference for pORERA-GFP vector or mention brand if it is commercial.	1	2	horticulturae8050412_perova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 119 and 276).	2	1	In line 113 replace “psoup” with pSOUP and “tobaccos” with tobacco plants.	1	2	horticulturae8050412_perova	1
+We have modified this part in revised manuscript (Line 121-125). This sentence has been changed to “Symbiotic medium was prepared by weighing 4.414 g MS, 20 g sucrose, 10 g glucose, 7 g agar into a conical flask, sterile water was fixed to 1 L. The medium was sterilized in an autoclave for 30 min and the temperature was lowered to 55℃, add 100 μl naphthalene acetic acid (0.1 mg L-1), 500 μl zeatin (1mg L-1), 1 mL folic acid (1mg L-1), 1 mL 6-BA (2mg L-1), 500 μl acetosyringone (0.1 mol L-1).” Point 15: In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	2	1	In line 117 and 267 replace “vacuuming penetration” with vacuum infiltration.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 129).	2	1	In line 118 define “symbiotic medium”.	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion, we had made correction carefully (Line 136).	2	1	In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction carefully (Line 138).	2	1	In line 129 replace “strains” and “strain” with lines and line.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 145).	2	1	In line 131 delete the period after experiments.	1	2	horticulturae8050412_perova	1
+"We had made correction in the manuscript (Line 156-157). The statements of “the MDA assay kit” were corrected as “the MDA assay kit (Jiancheng Bioengineering Institute, Nanjing, China).” Point 20: In line 154 add ""strain"" after receptor and mention kit brand Response 20: Thank you for your comment."	2	1	In line 138 Seedling should be lowercase.	1	2	horticulturae8050412_perova	1
+"We had made correction in the manuscript (Line 162 and 163). The statements of “the yeast Y187 receptor strain preparation and transformation kit” were corrected as “the yeast Y187 receptor strain preparation and transformation kit (Coolaber, Beijing, China).” Point 21: In line 169 add ""were done"" at the end."	2	1	In line 149 mention the kit brand.	1	2	horticulturae8050412_perova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 177).	2	1	In line 154 add “strain” after receptor and mention kit brand.	1	2	horticulturae8050412_perova	1
+Thank you for your comment, we had made correction in the manuscript (Line 270).	2	1	In line 169 add “were done” at the end. A phrase must have a verb.	1	2	horticulturae8050412_perova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 271).	2	1	In line 261 replace “tobaccos” with tobacco plants.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction carefully (Line 273).	2	1	In line 262 replace “were treatment by” with were treated with.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 301).	2	1	In line 264 replace “indicate” with indicated.	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction carefully (Line 327).	2	1	In line 292 “The” must be lowercase.	1	2	horticulturae8050412_perova	1
+Thank for your kindly suggestion, we had made correction carefully (Line 403).	2	1	In line 318 “We” must be lowercase.	1	2	horticulturae8050412_perova	1
+Thank you for your comment, we had made correction in the manuscript (Line 415).	2	1	In line 395 replace “it revealed” with revealed.	1	2	horticulturae8050412_perova	1
+We feel sorry for the unclear statement. In our study, we used the amino acid sequence of OfSPL11 to compute the protein model. Secondary structure predictions were conducted on the PRABI (http://www.prabi.fr/spip.php?page=sommaire) protein structure prediction server. The ExPASy (http://web.expasy.org) was used to analyze the molecular weight, theoretical isoelectric point, and other physiochemical properties of the translated protein. Similar methods of operation and efforts can be found in other articles (Islam, 2021). Once again,we sincerely thank you for the recommended method, which we will use in future studies.	2	1	Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?	1	2	horticulturae8050412_perova	1
+We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015).	2	1	Auhtors report that Actin was used as housekeeping gene. Do the authors have performed an efficiency test before using it? Do they tried to use otherHK genes?	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 130).	2	1	Line 124: generate instead of generate	1	2	horticulturae8050412_perova	1
+We have revised the manuscript according to your comments (Line 274-277). This sentence has been changed to “A. tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 3: Rephrase lines 317-318.	2	1	"Line 267-268: Please, check this sentence. It ends with ""then"".
+Reviewer_2	Author	1	5	0	2	Figure 5 caption: activates instead of activate
+Reviewer_2	Author	1	6	0	2	Line 302: plant leaves instead of plants leaves
+Reviewer_2	Author	1	7	0	2	Line 311: plant leaves instead of plants leaves
+Reviewer_2	Author	1	8	0	2	Line 315: maybe a semicolon after """"salt stress"""" is appropriate"""	1	2	horticulturae8050412_perova	1
+We are very sorry for our negligence, we had made correction in the manuscript (Line 366).	2	1	"Line 360: remove the space after ""assay"""	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion. We had made correction carefully (Line 381).	2	1	Figure 9 caption: measure instead of measuring	1	2	horticulturae8050412_perova	1
+Special thanks to you for your good suggestion, we had made correction in the manuscript (Line 398).	2	1	"Line 392: I suggest to change the sentence in the following way "".. to drive the expression of target genes.."""	1	2	horticulturae8050412_perova	1
+"Thank for your kindly suggestion. We have revised the manuscript according to your comments (Line 456). The statements of “participate in salt stress responses” were corrected as “participate in salt stress.” Point 12: Line 448: check the word ""redycing"" Response 12: Thank you for your comment."	2	1	"Line 446: ""...participate in salt stress"""	1	2	horticulturae8050412_perova	1
+Thank you for your comment. We are very sorry for our negligence. We had made correction carefully (Line 457). The word of “redycing” were corrected as “reducing.” 2 1	2	1	"Line 448: check the word ""redycing"""	1	2	horticulturae8050412_perova	1
+Thanks for your reminder. In our study, 66 children were taken part in our analysis. But we failed to get all children’s feces and urine sample every day. Thus, we collected sample number was less the involved 66 children.	2	1	Lines 116-118: The amounts of samples were confused. Although there were 66 children involved in this study, 66 food samples, 62 feces and 64 urine samples were collected.	1	2	ijerph19127332_makarova	1
+Thank you for your reminder. We have uniformed the units to “(mL/d)” Point 4.	2	1	Lines 176-178: Different units between manuscript and SM (figure S1).	1	2	ijerph19127332_makarova	1
+At present, the related content in manuscript has been revised to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Lines 209-211: Different SIR values between manuscript and figure 1.	1	2	ijerph19127332_makarova	1
+In the manuscript, the data is with outlier. However, we used the data without outlier to construct the figure. The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 5. At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.	1	2	ijerph19127332_makarova	1
+We have reorganized the SM figures and the manuscript content. The modifications are as follows: “The frequency distribution histograms show few outliers, most being high values (Figure S3 (a) and Figure S4). Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers. Other outliers may be due to the behavior of certain children, such as pica behavior or spending unusually large amounts of time playing in grassland. The medians of the SIR values after removing the high values (see Figure S3(b) and Figure 1) were taken as the final SIR values for children living in the studied e‒waste dismantling site in South China.” Point 7.	2	1	Lines 218 and 223: check the Figure 3 and figure S1.	1	2	ijerph19127332_makarova	1
+It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5). However, since negative SIR values are physically meaningless, the RI of the SIR for children living in e‒waste dismantling sites is 0‒730.4 mg/d. In this study, 95th per-centile values (383.3 mg/g) would be as recommended value.” Point 8.	2	1	Lines 253 and 377: different values between SIR and figure 3.	1	2	ijerph19127332_makarova	1
+It has been changed to “The 95th HQ values based on SIR mean value decreased in the order of Pb > As > Cr > Cu > Ni > Zn > Cd and all of them were below 1.” Point 9.	2	1	Lines 348-349: correct the order of HQs.	1	2	ijerph19127332_makarova	1
+We have revised all the figure captions in SM.	2	1	correct the figure caption in SM.	1	2	ijerph19127332_makarova	1
+Thanks for your advice, we have made the changes to the title following:” Estimation of Children’s Soil/Dust Ingestion Rates and Health Risk at E‑Waste Dismantling Area”	2	1	Title- ‘in’ should be used for larger areas, can you add the study locations? Otherwise replace ‘in’ with ‘at’Rephrase title so as not to repeat ‘and’ Answer:	1	2	ijerph19127332_makarova	1
+Thanks for your suggestion. We have introduced the sampling sites and added the following information: “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428 Point 3.	2	1	It was only mentioned towards the end – Discussion Answer: Thanks for your suggestion. Materials and Methods:I would like to see a better description of the study location.	1	2	ijerph19127332_makarova	1
+Thanks for your suggestion, we have sorted out the method part again, and now the modification is as follows: “Feces samples and food samples were pretreated in the same way. The digestion process was repeated until the sample becomes sticky.	2	1	Better explanation needed - why different processing for different samples?	1	2	ijerph19127332_makarova	1
+For this question, our answer is following: this data is from the Technical Guidelines for Soil Pollution Risk Assessment of Construction Land in China. The website address of the guidelines is https://www.mee.gov.cn/ywgz/fgbz/bz/bzwb/trhj/201912/W020191224560850148092.pdf	2	1	Line190 – why was 350d/y taken? Explain for better understanding	1	2	ijerph19127332_makarova	1
+For this problem, we conducted a questionnaire survey when collecting samples before the experiment, and all the guardians of the sample providers knew and agreed to the experiment.	2	1	Will like to see ethical approval for the project/research? Ethical approval from which institution? Were Consent/assent from participants obtained?	1	2	ijerph19127332_makarova	1
+Collecting samples followed the USEPA recommendation of a period of 28 hours from food to feces and urine. Thus, we cannot provide the data for each day. And we also revised the table 1 legend (removing the “each day”).	2	1	Results and Discussion: Table 1 – is it possible to list/ summarise the data for each day?	1	2	ijerph19127332_makarova	1
+For the convenience of you and readers, we have made full descriptions of some acronyms, and the modifications are as follows: “Therefore, in this study, the Best Tracer Method (BTM) was employed. This method has been used before”, “the Hazard Quotients (HQ) values between 1 and 10 indicate likely damage to human health”.	2	1	Line 294 – what is meaning of BTM, Would prefer re-introduction of the full meaning of acronyms in each sub-section Answer: Thank you for your suggestions.	1	2	ijerph19127332_makarova	1
+Thank you for your suggestions. We have added the description of sampling site in the materials and methods. We have replied to your first suggestion on Materials and Methods for the specific content to be added.	2	1	Line 320 -Also descript the study location in the method section Answer: Thank you for your suggestions.	1	2	ijerph19127332_makarova	1
+We have added the values for carcinogenic risk and non-carcinogenic risks in the support information.	2	1	Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.	1	2	ijerph19127332_makarova	1
+Thank you for your proposal. For this part, we have added to the methods section, and followed as: “The Hazard Quotients (HQ) is the ratio of daily intake dose of pollutants to reference dose, which is used to characterize the levels of human exposure to non-carcinogenic contaminants through a single pathway which represents the level of non‒carcinogenic risk”.	2	1	Line 346 – What is HQ? Also describe in the methods section Answer:	1	2	ijerph19127332_makarova	1
+Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities”	2	1	Line 368 – what is the meaning of ‘Whatever’? please rephrase Answer:	1	2	ijerph19127332_makarova	1
+Thank you for your advice. At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5). Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas. We have examined the tracer elements in the food. And when we calculated the SIR, we also used the parameter “Cfood” (equation 1) to analyze the food intake. In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR. Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue. At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd. The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas. Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Point 2.	2	1	Will like to see references on the metal concentrations at the e-waste recycling site.	1	2	ijerph19127332_makarova	1
+Thanks for your advice, and this explanation has been removed from the manuscript, it is changed to “this method analyzes the concentration of tracer elements in the soil to which children are exposed, the children's intake of food, their excreted feces and urine, and the content of the tracer element in the children’s food, feces, and urine.”	2	1	Page 2, lines 76-78: You can remove this sentence: a class of elements in the human body that are not easily absorbed by the human gastrointestinal tract and are also difficult to be transformed into other substances it’s not necessary Answer:	1	2	ijerph19127332_makarova	1
+; Shi, X.L. ; Wu, K.S. Human Body Burden of Heavy Metals and Health Consequences of Pb Exposure in Guiyu, an E-Waste Recycling Town in China. Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428” And we changed content as “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Point 3.	2	1	Page 3, lines 99-100: please, add references for the following information South China's economic conditions are better than in North China, but the environmental pollution is worse. The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.	1	2	ijerph19127332_makarova	1
+Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C). Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.	2	1	Freeze-drying conditions of feces. Page 3, lines 116 and 120: Please, add technical details about: lyophilization of food samples.	1	2	ijerph19127332_makarova	1
+Thank you for your suggestion. We have added the content we did not mention in this part, but the soil was not quartered, so there is no supplement. Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills. And 20 g of soil and as much dust (5-20 g) as possible were collected during the sampling process.” Point 5.	2	1	How much soil (kg) did you collect for each sample? Was the soil quartered? Page 3, lines 121-124: Please, add some information: How did you collect dust samples?	1	2	ijerph19127332_makarova	1
+Dried samples (1 g) were digested to evaporate at low temperatures of 55°C on a heating plate with 3 mL concentrated nitric acid, 3 mL hydrogen fluoride, and 1 mL perchloric acid (HNO3‒HF‒HClO4). Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2). Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min. The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C. The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3. The conditions of microwave digestion are the same as mentioned above. Finally, the digest was analyzed by HR‒ICP‒MS. Dried soil and dust samples (0.5 g) were digested as same as feces and food samples on a heating plate by HNO3‒HF‒HClO4. And then samples were microwaved with HNO3‒H2O2. The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y. In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn. The standard curve of concentration was used to determine the sample concentration was established by heavy metal standard (all standards were from The Nonferrous Metals Society of China).” Point 4. Thanks for your advice! In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way. The digestion process is repeated until the sample becomes sticky.	2	1	Lines 137-138: Please, modify as suggested The supernatant was then extracted and analyzed by HR‒ICP‒MS to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y, Answer: Thanks for your advice!	1	2	ijerph19127332_makarova	1
+Thank you for your correction. We have added (a, b, c, ect) to Figure 1 and Figure S1,2,3,4. The title of Figure 1 has been changed to “Figure 1. Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h).” The title of Figure S1 has been changed to “Fig. S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”. The title of Figure S2 has been changed to “Fig. S2 Histogram and basic statistical parameters of daily food ingestion (a) (g/d, ww) and feces (b) (g/d, dw) and urine (c) (mL/d) excretion for investigated child population.”. The title of Figure S3 has been changed to “Fig. S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”. The title of Figure S4 has been changed to “Fig. S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.	2	1	Please, add labels (a, b, c, etc.) in each figure and update the captions. Also in the supplementary. Please, check captions of figures S2 and 3 that are identical.	1	2	ijerph19127332_makarova	1
+Thanks for your reminder. In our study, 66 children were taken part in our analysis. But we failed to get all children’s feces and urine sample every day. Thus, we collected sample number was less the involved 66 children.	2	1	Lines 116-118: The amounts of samples were confused. Although there were 66 children involved in this study, 66 food samples, 62 feces and 64 urine samples were collected.	1	2	ijerph19127332_perova	1
+Thank you for your reminder. We have uniformed the units to “(mL/d)” Point 4.	2	1	Lines 176-178: Different units between manuscript and SM (figure S1).	1	2	ijerph19127332_perova	1
+At present, the related content in manuscript has been revised to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Lines 209-211: Different SIR values between manuscript and figure 1.	1	2	ijerph19127332_perova	1
+In the manuscript, the data is with outlier. However, we used the data without outlier to construct the figure. The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 5. At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.	1	2	ijerph19127332_perova	1
+We have reorganized the SM figures and the manuscript content. The modifications are as follows: “The frequency distribution histograms show few outliers, most being high values (Figure S3 (a) and Figure S4). Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers. Other outliers may be due to the behavior of certain children, such as pica behavior or spending unusually large amounts of time playing in grassland. The medians of the SIR values after removing the high values (see Figure S3(b) and Figure 1) were taken as the final SIR values for children living in the studied e‒waste dismantling site in South China.” Point 7.	2	1	Lines 218 and 223: check the Figure 3 and figure S1.	1	2	ijerph19127332_perova	1
+It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5). However, since negative SIR values are physically meaningless, the RI of the SIR for children living in e‒waste dismantling sites is 0‒730.4 mg/d. In this study, 95th per-centile values (383.3 mg/g) would be as recommended value.” Point 8.	2	1	Lines 253 and 377: different values between SIR and figure 3.	1	2	ijerph19127332_perova	1
+It has been changed to “The 95th HQ values based on SIR mean value decreased in the order of Pb > As > Cr > Cu > Ni > Zn > Cd and all of them were below 1.” Point 9.	2	1	Lines 348-349: correct the order of HQs.	1	2	ijerph19127332_perova	1
+We have revised all the figure captions in SM.	2	1	correct the figure caption in SM.	1	2	ijerph19127332_perova	1
+Thanks for your advice, we have made the changes to the title following:” Estimation of Children’s Soil/Dust Ingestion Rates and Health Risk at E‑Waste Dismantling Area”	2	1	Title- ‘in’ should be used for larger areas, can you add the study locations? Otherwise replace ‘in’ with ‘at’Rephrase title so as not to repeat ‘and’ Answer:	1	2	ijerph19127332_perova	1
+Thanks for your suggestion. We have introduced the sampling sites and added the following information: “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428 Point 3.	2	1	I would like to see a better description of the study location. It was only mentioned towards the end – Discussion Answer: Thanks for your suggestion.	1	2	ijerph19127332_perova	1
+Thanks for your suggestion, we have sorted out the method part again, and now the modification is as follows: “Feces samples and food samples were pretreated in the same way. The digestion process was repeated until the sample becomes sticky.	2	1	Better explanation needed - why different processing for different samples?	1	2	ijerph19127332_perova	1
+For this question, our answer is following: this data is from the Technical Guidelines for Soil Pollution Risk Assessment of Construction Land in China. The website address of the guidelines is https://www.mee.gov.cn/ywgz/fgbz/bz/bzwb/trhj/201912/W020191224560850148092.pdf Will like to see ethical approval for the project/research? Ethical approval from which institution? Were Consent/assent from participants obtained? For this problem, we conducted a questionnaire survey when collecting samples before the experiment, and all the guardians of the sample providers knew and agreed to the experiment.	2	1	Line190 – why was 350d/y taken? Explain for better understanding Answer:	1	2	ijerph19127332_perova	1
+Collecting samples followed the USEPA recommendation of a period of 28 hours from food to feces and urine. Thus, we cannot provide the data for each day. And we also revised the table 1 legend (removing the “each day”).	2	1	Table 1 – is it possible to list/ summarise the data for each day?	1	2	ijerph19127332_perova	1
+For the convenience of you and readers, we have made full descriptions of some acronyms, and the modifications are as follows: “Therefore, in this study, the Best Tracer Method (BTM) was employed. This method has been used before”, “the Hazard Quotients (HQ) values between 1 and 10 indicate likely damage to human health”.	2	1	Line 294 – what is meaning of BTM, Would prefer re-introduction of the full meaning of acronyms in each sub-section Answer: Thank you for your suggestions.	1	2	ijerph19127332_perova	1
+Thank you for your suggestions. We have added the description of sampling site in the materials and methods. We have replied to your first suggestion on Materials and Methods for the specific content to be added.	2	1	Line 320 -Also descript the study location in the method section Answer: Thank you for your suggestions.	1	2	ijerph19127332_perova	1
+We have added the values for carcinogenic risk and non-carcinogenic risks in the support information.	2	1	Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.	1	2	ijerph19127332_perova	1
+Thank you for your proposal. For this part, we have added to the methods section, and followed as: “The Hazard Quotients (HQ) is the ratio of daily intake dose of pollutants to reference dose, which is used to characterize the levels of human exposure to non-carcinogenic contaminants through a single pathway which represents the level of non‒carcinogenic risk”.	2	1	Line 346 – What is HQ? Also describe in the methods section Answer:	1	2	ijerph19127332_perova	1
+Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities”	2	1	Line 368 – what is the meaning of ‘Whatever’? please rephrase Answer:	1	2	ijerph19127332_perova	1
+Thank you for your advice. At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5). Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas. We have examined the tracer elements in the food. And when we calculated the SIR, we also used the parameter “Cfood” (equation 1) to analyze the food intake. In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR. Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue. At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd. The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas. Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Point 2.	2	1	Will like to see references on the metal concentrations at the e-waste recycling site.	1	2	ijerph19127332_perova	1
+Thanks for your advice, and this explanation has been removed from the manuscript, it is changed to “this method analyzes the concentration of tracer elements in the soil to which children are exposed, the children's intake of food, their excreted feces and urine, and the content of the tracer element in the children’s food, feces, and urine.”	2	1	Page 2, lines 76-78: You can remove this sentence: a class of elements in the human body that are not easily absorbed by the human gastrointestinal tract and are also difficult to be transformed into other substances it’s not necessary Answer:	1	2	ijerph19127332_perova	1
+; Shi, X.L. ; Wu, K.S. Human Body Burden of Heavy Metals and Health Consequences of Pb Exposure in Guiyu, an E-Waste Recycling Town in China. Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428” And we changed content as “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Point 3.	2	1	Page 3, lines 99-100: please, add references for the following information South China's economic conditions are better than in North China, but the environmental pollution is worse. The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.	1	2	ijerph19127332_perova	1
+Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C). Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.	2	1	Page 3, lines 116 and 120: Please, add technical details about:lyophilization of food samples. Freeze-drying conditions of feces.	1	2	ijerph19127332_perova	1
+Thank you for your suggestion. We have added the content we did not mention in this part, but the soil was not quartered, so there is no supplement. Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills. And 20 g of soil and as much dust (5-20 g) as possible were collected during the sampling process.” Point 5.	2	1	Page 3, lines 121-124:Please, add some information: How did you collect dust samples? How much soil (kg) did you collect for each sample? Was the soil quartered?	1	2	ijerph19127332_perova	1
+Dried samples (1 g) were digested to evaporate at low temperatures of 55°C on a heating plate with 3 mL concentrated nitric acid, 3 mL hydrogen fluoride, and 1 mL perchloric acid (HNO3‒HF‒HClO4). Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2). Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min. The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C. The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3. The conditions of microwave digestion are the same as mentioned above. Finally, the digest was analyzed by HR‒ICP‒MS. Dried soil and dust samples (0.5 g) were digested as same as feces and food samples on a heating plate by HNO3‒HF‒HClO4. And then samples were microwaved with HNO3‒H2O2. The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y. In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn. The standard curve of concentration was used to determine the sample concentration was established by heavy metal standard (all standards were from The Nonferrous Metals Society of China).” Point 4. Thanks for your advice! In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way. The digestion process is repeated until the sample becomes sticky.	2	1	Lines 131-135: Please, modify as suggested Crushed food samples (1 g) were microwaved with concentrated nitric acid and hydrogen peroxide (HNO3‒H2O2). The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Lines 137-138: Please, modify as suggested The supernatant was then extracted and analyzed by HR‒ICP‒MS to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y, Answer: Thanks for your advice!	1	2	ijerph19127332_perova	1
+Thank you for your correction. We have added (a, b, c, ect) to Figure 1 and Figure S1,2,3,4. The title of Figure 1 has been changed to “Figure 1. Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h).” The title of Figure S1 has been changed to “Fig. S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”. The title of Figure S2 has been changed to “Fig. S2 Histogram and basic statistical parameters of daily food ingestion (a) (g/d, ww) and feces (b) (g/d, dw) and urine (c) (mL/d) excretion for investigated child population.”. The title of Figure S3 has been changed to “Fig. S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”. The title of Figure S4 has been changed to “Fig. S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.	2	1	Please, add labels (a, b, c, etc.) in each figure and update the captions. Also in the supplementary. Please, check captions of figures S2 and 3 that are identical.	1	2	ijerph19127332_perova	1
+Thank you for this suggestion. While it would have been interesting to explore this further, our study focuses on retrospectively assessing and comparing disease progression within subgroups of patients receiving TOC. Patients not receiving TOC would have likely had a different presentation, progression and their follow-up would be slightly out of the scope of our observational retrospective study. When we compared patients who received TOC with the 86 severe patients who did not receive TOC (excluded from this analysis), the TOC group was far more severe, so there is no control group that could be matched. In addition, these patients did not have control CT scans.	2	1	The impact of Tocilizumab on radiological changes can not be adequately analyzed without a control group not receiving the drug.	1	2	jcm11051247_makarova	1
+Following this suggestion, we have included in the analysis a calculation according to the FiO2. This has a cut-off which correlates to the oxygen flow cut-off.	2	1	The proposed timing of Tocilizumab administration based on oxygen flow rate is interesting but undoubtedly requires a more detailed statistical analysis to convince the readers.	1	2	jcm11051247_makarova	1
+This is an interesting point. While knowledge of variants is of great importance from an epidemiologic point of view as they have different transmission rates and can cause diseases of different severity, the mechanism of the acute inflammatory response in severe infection seems to be similar regardless the strain. While we did not perform specific sequencing of the strain for every patient as this wouldn’t have been feasible, patients were included before the Delta variant emerged and we expect most of them to have had the initial D614G and UK B.1.1.7. Nonetheless, we have included a comment addressing this aspect in our limitations section to highlight this area of uncertainty.	2	1	Materials and Methods 2.1. Study design and population If there are mutations or differences in viral strains during the course of the study, should differences in response to TOC and timing of administration depending on the strain be considered?	1	2	jcm11051247_makarova	1
+We agree with the referee that the reasons for the exclusions were unclear and have rephrased accordingly. Addresed in text: „All those excluded were not administered TOC either because of contraindications or TOC supply gaps.” Point 3: 3.	2	1	Results 86 cases are excluded from 187 cases, which is too many. What is the reason?	1	2	jcm11051247_makarova	1
+We added the data using FiO2 ROC curve. The area under the curve for FiO2 ROC curve was smaller than the area under the curve for oxygen flow. In addition, the cut-off value for FiO2 corresponded to the cut-off for the oxygen flow rate, so the two groups in Table 3 would be similar.	2	1	Results When comparing different oxygen delivery methods, isn't it more appropriate to compare FiO2 rather than oxygen flow rate?	1	2	jcm11051247_makarova	1
+We added this information also in Table 3. We analysed FiO2 (depending on the method of oxygen delivery - nasal canulae, HNF, MV), as you suggested and the cut-off value of FiO2 corresponded to the cut-off for the oxygen flow rate, so the two groups in Table 3 would be similar.	2	1	Results The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).	1	2	jcm11051247_makarova	1
+Addressed in text and in Table 3.	2	1	The oxygen flow rate of 13 L/min measured by ROC as a cutoff value for TOC administration should be presented by FiO2.	1	2	jcm11051247_makarova	1
+Thank you for pointing this out, we have changed Table 1 accordingly and we removed the imaging data, which as you mentioned, was the same as “before TOC” in Table 2.	2	1	Comparative radiologic changes before and after TOC administration In Table 2, about the change in CT findings in fatal and non-fatal patients before and after TOC administration, isn't it the same as the difference in imaging findings between patients who survive after severe illness and those who do not, regardless of TOC?	1	2	jcm11051247_makarova	1
+Addressed, see previous comments and answers.	2	1	Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.	1	2	jcm11051247_makarova	1
+separation of sections and subheadings, revising all the text, adding paragraphs & references, adding/changing Tables & Figures): 1) Abstract was revised.	2	1	Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg. prediction mechanism, prediction accuracy, etc.) is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.	1	2	land11030363_makarova	1
+Sorry for the technical error. The sample set was divided into two subsets (75% and 25%) for calibration (86 samples) and validation (28 samples).	2	1	When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio. There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?	1	2	land11030363_makarova	1
+The sections were separated as recommended.	2	1	Should the writing of the results and discussion sections be separated? Please think carefully.	1	2	land11030363_makarova	1
+The comment was fully considered: the PCA and partial correlation analysis was adopted form the recommended study (e.g., Cheng et al., 2019). Correspondingly, separate subheadings (with new Table 4 and Figure 4) were added. Generally it was considered in Introduction and Materials & Methods, Results and Discussion sections. Such approach helped to analyze the underlying mechanisms.	2	1	In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).	1	2	land11030363_makarova	1
+It was corrected to “modeling”.	2	1	Line 104, 195, 223, 544: Is it “modelling” or “modeling”? The whole text should be unified.	1	2	land11030363_makarova	1
+It was corrected. pH was measured in water solution. (Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?	2	1	Lines 145-146: “, soil reaction (pH) in 1:1 soil to KCl solution” is not clearly expressed.	1	2	land11030363_makarova	1
+It was revised as recommended. (Line: 244-249) Line 174: “…resulting in 12 spectra”.	2	1	Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence? Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources. What does the controlled laboratory environment you describe look like?	1	2	land11030363_makarova	1
+The text was revised. Each single measurement was based on internally averaged of 50 spectra (3 times). Each sample was measured from four positions (90°rotating the Petri dish each time). Consequently, 4 rotation x 3 time = 12 spectra were obtained. It is one of the commonly used method and several references are available. Line;252-259 Line 176: “…resulted 50 spectra…”.	2	1	Line 174: “…resulting in 12 spectra”. It is not clear how the 12 spectra were obtained here.	1	2	land11030363_makarova	1
+See previous comment.	2	1	Line 176: “…resulted 50 spectra…”. It is also not clear how 50 spectra were obtained.	1	2	land11030363_makarova	1
+The whole paragraph was revised. Line: 262-269. We would like to note that in practice a large variety of pre-processing techniques are used with different gap and moving window size to improve prediction quality (Luce et al. 2017, Mammadov at al. 2020). Yet, the quality of prediction still depends on several factors (measurement method, used device etc.).	2	1	Line 190: “…moving window sizes (mainly 7, 9 and 11)”. Need to use 3 different moving window sizes? It undoubtedly increases your experimental workload, and you do not explain it later. As far as I know, most articles use only one moving window size.	1	2	land11030363_makarova	1
+The text was revised, see previous comment.	2	1	Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”. Do these two places mean the same thing?	1	2	land11030363_makarova	1
+It was revised as “change in pH, temperature, precipitation by elevation”. Line:334-337.	2	1	Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.	1	2	land11030363_makarova	1
+Corrected to “soil type”.	2	1	Line 264: “…, and hence parent material.” It’s not clear here.	1	2	land11030363_makarova	1
+Both methods are used to characterize relations between soil properties and spectra. Based on our experience, for current study (or similar studies) relations among soil properties, and between PC1 and soil properties are well characterized by Spearman, and Pearson is more relevant to the relations between spectra and soil properties.	2	1	Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?	1	2	land11030363_makarova	1
+Line:515-518.	2	1	Lines 325-328: “Regardless of… spectra.” Is there a necessary connection between these two sentences? The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.	1	2	land11030363_makarova	1
+Line:43, 440 Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here.	2	1	Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.	1	2	land11030363_makarova	1
+It was modified. Line: 438-441 Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	2	1	Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here. And all in Table 3 are “10-bands”.	1	2	land11030363_makarova	1
+Line: 696-699 Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	2	1	Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	1	2	land11030363_makarova	1
+Line:702-705 Line 409: Note the singular and plural in the sentence.	2	1	Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	1	2	land11030363_makarova	1
+Corrected to “CaCO3”. Line: 750-752 Line 455: “soil mineralogy”?	2	1	Line 449, 479: “CO3”?	1	2	land11030363_makarova	1
+It was replaced with “clay mineralogy”.	2	1	Line 455: “soil mineralogy”?	1	2	land11030363_makarova	1
+Line 736-740 Line 471, 480, 520: “(r = 0.4*)” “(r =-0.52*)” “(r = 0.40*)”, etc.	2	1	Lines 456-457: What does this sentence do here?	1	2	land11030363_makarova	1
+Line:759-762 Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what?	2	1	Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.	1	2	land11030363_makarova	1
+It was revised. It was related to the contribution of the basic soil properties (e.g. controlling factor CaCO3, Fe, clay) to the prediction of M3 extractable elements Line:780-786.	2	1	Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what? Readers may be more interested in your analysis and interpretation.	1	2	land11030363_makarova	1
+Corrected or revised. Line:786-789 Line 523: “correspond absorption” should be “correspond to absorption”.	2	1	Line 520: The correlation between K content and P content is not very high (r = 0.40*), but their corresponding VIP patterns are “identical”, how do you explain it?	1	2	land11030363_makarova	1
+It was edited. Line:790-791 Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence?	2	1	Line 523: “correspond absorption” should be “correspond to absorption”.	1	2	land11030363_makarova	1
+Partial correlation analysis showed that the contribution of Fe was more important than that of SOC though the moderate correlation existed between them. Line: 786-791 Lines 541-543: Is this sentence appropriate here?	2	1	Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence? Is VIP peak related to Fe or SOC content?	1	2	land11030363_makarova	1
+We have followed the instruction. The change was marked with the red letter.	4	1	Many thanks for your response. The manuscript has been largely improved. Modified content should be marked in the revised manuscript for easy review by reviewers.	3	2	land11030363_makarova	1
+The abstract was revised to show all key results.	4	1	The abstract could be more concise.	3	2	land11030363_makarova	1
+Introduction was modified.	4	1	The introduction part can highlight the key points and innovations of this paper.	3	2	land11030363_makarova	1
+The manuscript was checked for missing articles and edited.	4	1	The definite article “the” is missing in many places in the whole article. please check it.	3	2	land11030363_makarova	1
+We reduced the number of parentheses significantly.	4	1	Too many parentheses are used in the essay, which is not recommended in writing.	3	2	land11030363_makarova	1
+It was removed.	4	1	Line 135: “impairment water resources” Is this expression appropriate?	3	2	land11030363_makarova	1
+Soory for the technical error. In the previous version, pH values were related to the determination in KCL solution. It was replaced with pH values in H2O water.	4	1	Table 1: The pH values are different from those in the previous version.	3	2	land11030363_makarova	1
+It was edited both in the text and tables.	4	1	Attention should be paid to the number of decimal places in all numbers throughout the text.	3	2	land11030363_makarova	1
+It was corrected.	4	1	Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit. Please check the format of all references.	3	2	land11030363_makarova	1
+Thanks. The conclusions were rewritten to reflect the key results.	4	1	The conclusion is too long and does not highlight the key results.	3	2	land11030363_makarova	1
+We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. New subheading (3.2. Land use effect on soil properties) was added. New Table 4 and Figure 4 were added. Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear). We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of section and subheadings, revising all the text, adding paragraphs and references, adding or changing Tables and Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. a) Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. b) New subheading (3.2. Land use effect on soil properties) was added. It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1. c). New Table 4 and Figure 4 were added. Table 4 is related to partial correlation analysis with consideration of soil properties, and Figure 4 is related to the principal component analysis (biplot). d) Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 6) Discussion with new 3 subheading was significantly expanded and linked to the references and the novelty was highlighted. 7) Conclusions of the study was modified to show its practical outcomes for the local and international readers. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Due to the separating the Results and Discussion section, overlapping of some comments made by the reviewers and associated changes modified all the text of the manuscript was modified. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear).	2	1	The novelty of the study should be clearly stated in the Introduction section after the objectives are formulated.	1	2	land11030363_makarova	1
+The section was revised and information regarding solution standards and detection limit was added. (M&M. subheading 2.2). Chemical analyses and descriptive statistics of the results b) It is necessary to explain the method of sampling using the appropriate literature.	2	1	"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses. Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments."	1	2	land11030363_makarova	1
+The sampling scheme was explained as recommended (M&M, subheading 2.1). In general, sampling scale changes between 1:12 000 and 1:250 000 (Soil Survey Staff, USDA NRCS 2016). In this pioneering study area, soil properties are highly variable attributed to topography, land use history and land tenure ownerships (mainly 0.2-0.3 hectares and fragmented shrubbery areas) and geological setting that makes difficult to apply regular sampling scheme. Therefore, the sampling locations were randomly (irregularly) designated, yet cover variations in land use, topography, geological substrate and the erosive state of soil continuum thereby resulted in 114 samples from 525 ha used under four land use types (1 sample per ~ 5 hectares).	2	1	b) It is necessary to explain the method of sampling using the appropriate literature. Is the number of samples sufficient for the purpose of the research? Please explain.	1	2	land11030363_makarova	1
+Two references were added. (Subheading 2.1) The “Conclusions section” is not very successful concerning the presentation of results.	2	1	c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned. Reference to formal analytical methods must be made in a scientific manner. The writing language used must be different from the way an analyst mentions it in the lab.	1	2	land11030363_makarova	1
+As mentioned above, we restructured the introduction section and included new paragraphs to illuminate the importance of the study. Special focus was given to clarifying prediction mechanisms for the studied soil properties and land use effect on soil properties.	2	1	The “Introduction section ” should be reworked, highlighting the relevance of this topic worldwide. It would be wise to give more international flavor in the sections of Introduction and Discussion.	1	2	land11030363_makarova	1
+The suggested study was reviewed and included to the reference list. It helped us to explain the results of our study (e.g. prediction of Fe and Cd using basic soil properties, clay mineralogy and (soil type) and micro-nutrients).	2	1	In addition, it needs support with more and more up-to-date recent articles. I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research,https://doi.org/10.1007/s11356-021-14673-0	1	2	land11030363_makarova	1
+Response: We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of section and subheadings, revising all the text, adding paragraphs and references, adding or changing Tables and Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. a) Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. b) New subheading (3.2. Land use effect on soil properties) was added. It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1. c). New Table 4 and Figure 4 were added. Table 4 is related to partial correlation analysis with consideration of soil properties, and Figure 4 is related to the principal component analysis (biplot). d) Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 6) Discussion with new 3 subheading was significantly expanded and linked to the references and the novelty was highlighted. 7) Conclusions of the study was modified to show its practical outcomes for the local and international readers. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Due to the separating the Results and Discussion section, overlapping of some comments made by the reviewers and associated changes modified all the text of the manuscript was modified. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear).	2	1	This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models. Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear. In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are. The abstract and conclusion parts lack conciseness and prominence. Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg. prediction mechanism, prediction accuracy, etc.) is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.	1	2	land11030363_perova	1
+Sorry for the technical error. The sample set was divided into two subsets (75% and 25%) for calibration (86 samples) and validation (28 samples).	2	1	When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio. There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?	1	2	land11030363_perova	1
+The sections were separated as recommended.	2	1	Should the writing of the results and discussion sections be separated? Please think carefully.	1	2	land11030363_perova	1
+The comment was fully considered: the PCA and partial correlation analysis was adopted form the recommended study (e.g., Cheng et al., 2019). Correspondingly, separate subheadings (with new Table 4 and Figure 4) were added. Generally it was considered in Introduction and Materials & Methods, Results and Discussion sections. Such approach helped to analyze the underlying mechanisms.	2	1	During the discussion, the reasons behind the experimental results should be analyzed, not only from the correlation between the sample data but also cannot be explained by relying too much on the VIP score curve. In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide). This may help to analyze the underlying mechanism.	1	2	land11030363_perova	1
+It was corrected	2	1	Line 104, 195, 223, 544: Is it “modelling” or “modeling”? The whole text should be unified.	1	2	land11030363_perova	1
+pH was measured in water solution.	2	1	Lines 145-146: ��, soil reaction (pH) in 1:1 soil to KCl solution” is not clearly expressed.	1	2	land11030363_perova	1
+It was revised as recommended.	2	1	Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence? Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources. What does the controlled laboratory environment you describe look like?	1	2	land11030363_perova	1
+The text was revised. Each single measurement was based on internally averaged of 50 spectra (3 times). Each sample was measured from four positions (90°rotating the Petri dish each time). Consequently, 4 rotation x 3 time = 12 spectra were obtained. It is one of the commonly used method and several references are available.	2	1	Line 174: “…resulting in 12 spectra”. It is not clear how the 12 spectra were obtained here.	1	2	land11030363_perova	1
+See previous comment.	2	1	Line 176: “…resulted 50 spectra…”. It is also not clear how 50 spectra were obtained.	1	2	land11030363_perova	1
+The whole paragraph was revised. Line: 262-269. We would like to note that in practice a large variety of pre-processing techniques are used with different gap and moving window size to improve prediction quality (Luce et al. 2017, Mammadov at al. 2020). Yet, the quality of prediction still depends on several factors (measurement method, used device etc.).	2	1	Line 190: “…moving window sizes (mainly 7, 9 and 11)”. Need to use 3 different moving window sizes? It undoubtedly increases your experimental workload, and you do not explain it later. As far as I know, most articles use only one moving window size.	1	2	land11030363_perova	1
+The text was revised, see previous comment.	2	1	Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”. Do these two places mean the same thing?	1	2	land11030363_perova	1
+It was revised as “change in pH, temperature, precipitation by elevation”. Line:334-337.	2	1	Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.	1	2	land11030363_perova	1
+Corrected to “soil type”.	2	1	Line 264: “…, and hence parent material.” It’s not clear here.	1	2	land11030363_perova	1
+Both methods are used to characterize relations between soil properties and spectra. Based on our experience, for current study (or similar studies) relations among soil properties, and between PC1 and soil properties are well characterized by Spearman, and Pearson is more relevant to the relations between spectra and soil properties.	2	1	Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?	1	2	land11030363_perova	1
+Line:515-518.	2	1	Lines 325-328: “Regardless of… spectra.” Is there a necessary connection between these two sentences? The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.	1	2	land11030363_perova	1
+Line:43, 440 Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here.	2	1	Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.	1	2	land11030363_perova	1
+It was modified. Line: 438-441 Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	2	1	Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here. And all in Table 3 are “10-bands”.	1	2	land11030363_perova	1
+Line: 696-699 Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	2	1	Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	1	2	land11030363_perova	1
+Line:702-705 Line 409: Note the singular and plural in the sentence.	2	1	Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	1	2	land11030363_perova	1
+Corrected to “CaCO3”. Line: 750-752 Line 455: “soil mineralogy”?	2	1	Line 449, 479: “CO3”?	1	2	land11030363_perova	1
+It was replaced with “clay mineralogy”.	2	1	Line 455: “soil mineralogy”?	1	2	land11030363_perova	1
+Line 736-740 Line 471, 480, 520: “(r = 0.4*)” “(r =-0.52*)” “(r = 0.40*)”, etc.	2	1	Lines 456-457: What does this sentence do here?	1	2	land11030363_perova	1
+Line:759-762 Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what?	2	1	Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.	1	2	land11030363_perova	1
+It was revised. It was related to the contribution of the basic soil properties (e.g. controlling factor CaCO3, Fe, clay) to the prediction of M3 extractable elements Line:780-786.	2	1	Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what? Readers may be more interested in your analysis and interpretation.	1	2	land11030363_perova	1
+Corrected or revised. Line:786-789 Line 523: “correspond absorption” should be “correspond to absorption”.	2	1	Line 520: The correlation between K content and P content is not very high (r = 0.40*), but their corresponding VIP patterns are “identical”, how do you explain it?	1	2	land11030363_perova	1
+It was edited. Line:790-791 Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence?	2	1	Line 523: “correspond absorption” should be “correspond to absorption”.	1	2	land11030363_perova	1
+Partial correlation analysis showed that the contribution of Fe was more important than that of SOC though the moderate correlation existed between them. Line: 786-791 Lines 541-543: Is this sentence appropriate here?	2	1	Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence? Is VIP peak related to Fe or SOC content?	1	2	land11030363_perova	1
+edited	2	1	Lines 541-543: Is this sentence appropriate here?	1	2	land11030363_perova	1
+The abstract was revised to show all key results.	4	1	The abstract could be more concise.	3	2	land11030363_perova	1
+Introduction was modified.	4	1	The introduction part can highlight the key points and innovations of this paper.	3	2	land11030363_perova	1
+The manuscript was checked for missing articles and edited.	4	1	The definite article “the” is missing in many places in the whole article. please check it.	3	2	land11030363_perova	1
+We reduced the number of parentheses significantly.	4	1	Too many parentheses are used in the essay, which is not recommended in writing.	3	2	land11030363_perova	1
+It was removed.	4	1	Line 135: “impairment water resources” Is this expression appropriate?	3	2	land11030363_perova	1
+Soory for the technical error. In the previous version, pH values were related to the determination in KCL solution. It was replaced with pH values in H2O water.	4	1	Table 1: The pH values are different from those in the previous version.	3	2	land11030363_perova	1
+It was edited both in the text and tables.	4	1	Attention should be paid to the number of decimal places in all numbers throughout the text.	3	2	land11030363_perova	1
+It was corrected.	4	1	Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit. Please check the format of all references.	3	2	land11030363_perova	1
+Thanks. The conclusions were rewritten to reflect the key results.	4	1	The conclusion is too long and does not highlight the key results.	3	2	land11030363_perova	1
+We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of sections and subheadings, revising all the text, adding paragraphs & references, adding/changing Tables & Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. New subheading (3.2. Land use effect on soil properties) was added. New Table 4 and Figure 4 were added. Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript.	2	1	The novelty of the study should be clearly stated in the Introduction section after the objectives are formulated.	1	2	land11030363_perova	1
+The section was revised and information regarding solution standards and detection limit was added. (M&M. subheading 2.2). Chemical analyses and descriptive statistics of the results b) It is necessary to explain the method of sampling using the appropriate literature.	2	1	There is not adequate information regarding quality control of the chemical analyses. Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.	1	2	land11030363_perova	1
+The sampling scheme was explained as recommended (M&M, subheading 2.1). In general, sampling scale changes between 1:12 000 and 1:250 000 (Soil Survey Staff, USDA NRCS 2016). In this pioneering study area, soil properties are highly variable attributed to topography, land use history and land tenure ownerships (mainly 0.2-0.3 hectares and fragmented shrubbery areas) and geological setting that makes difficult to apply regular sampling scheme. Therefore, the sampling locations were randomly (irregularly) designated, yet cover variations in land use, topography, geological substrate and the erosive state of soil continuum thereby resulted in 114 samples from 525 ha used under four land use types (1 sample per ~ 5 hectares).	2	1	It is necessary to explain the method of sampling using the appropriate literature. Is the number of samples sufficient for the purpose of the research? Please explain.	1	2	land11030363_perova	1
+Two references were added. (Subheading 2.1) The “Conclusions section” is not very successful concerning the presentation of results.	2	1	Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned. Reference to formal analytical methods must be made in a scientific manner. The writing language used must be different from the way an analyst mentions it in the lab.	1	2	land11030363_perova	1
+As mentioned above, we restructured the introduction section and included new paragraphs to illuminate the importance of the study. Special focus was given to clarifying prediction mechanisms for the studied soil properties and land use effect on soil properties.	2	1	The “Introduction section ” should be reworked, highlighting the relevance of this topic worldwide. It would be wise to give more international flavor in the sections of Introduction and Discussion.	1	2	land11030363_perova	1
+The suggested study was reviewed and included to the reference list. It helped us to explain the results of our study (e.g. prediction of Fe and Cd using basic soil properties, clay mineralogy and (soil type) and micro-nutrients).	2	1	In addition, it needs support with more and more up-to-date recent articles. I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research,https://doi.org/10.1007/s11356-021-14673-0	1	2	land11030363_perova	1
+We agree, and we have noted this point in the revised study limitations section.	2	1	The authors admit that, line 367, younger people were overrepresented, but to me, it was that older persons, especially over age 70, who were somehow missed in the survey sampling process. The study offers little to say about long covid among the elderly.	1	2	life12060901_makarova	1
+Interesting point, which we have added to the revised Conclusion and Implications section.	2	1	The authors consider a number of factors associated with long COVID and psychological symptoms. I would suggest another factor - frustration with not getting well.	1	2	life12060901_makarova	1
+Thank you for the kind response.	2	1	I accept the manuscript for its publication in this journal.	1	2	life12060901_makarova	1
+Please see our comments to each of the listed issues below.	2	1	However, I see several issues with the ending of this paper. I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential. However, what I disliked was the interpretation and discussion of the empirical results. Frankly speaking, the authors are overselling their results. I found two major issues in the discussion of this paper: Authors response:	1	2	life12060901_makarova	1
+The first mentioned statement has been removed altogether in the revised manuscript. The second statement has been modified in accordance with the reviewer’s view (see revised section 4.3).	2	1	The authors said in the statistic section that “statistical significance was set at p<0.05.” (page 5, line 212). However, later they wrote “The descriptively higher proportion of females with long COVID bordered statistical significance (p=0.05)” page 5, line 230) and “However, the different proportions of men and women with long COVID (22% and 33%, respectively) bordered towards statistical significance […]” (page 10, line 318). As a statistician, I felt very mad about this, because the results speak a different language, this is non-significant result.	1	2	life12060901_makarova	1
+Thank you for pointing this out. We have removed the relevant section in accordance with this guidance.	2	1	The authors wrote later in the conclusion section “[…] but a non-significant trend was found for the association between female gender and long COVID.” (page 11, line 389). This is a “no-go” in scientific research, the interpretation of “trends” resulted from non-significant results is strongly misleading. Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05). That is the true story. Hence, I urge the authors not to mislead readers and to re-write these sections.	1	2	life12060901_makarova	1
+Thank you for pointing this out, we have revised the sentence (see section 4.1).	2	1	The next major issue targets the interpretation of interaction effects. The authors did three models with three different outcomes, namely psychological distress, fatigue, and perceived stress. In the results section, only two (and not three!) interaction effects became significant (psychological distress: p<0.001, fatigue: p<0.05, and perceived stress: p=0.36, see Table 4, page 7, line 269). However, the authors wrote “While participants with long COVID generally perceived more psychological distress, fatigue, and stress than those without long COVID, differences were larger for men than for women” page 9, line 303), Authors’ response: Authors introduced an interaction effect long COVID x gender into the model in order to test their third hypothesis (“(iii) whether gender moderated the associations between long 119 COVID status and the health outcomes”, page 3, line 119).	1	2	life12060901_makarova	1
+We have revised the sentence; see section 4.4.	2	1	“[…] men’s perception of poorer health when having long COVID appears to include higher levels of psychological distress, fatigue and perceived stress” (page 11, line 363),	1	2	life12060901_makarova	1
+We have revised the sentence; see section 5.	2	1	and finally “Third, long COVID appears to have a stronger effect on men than on women” (page 11, line 401). Again, the authors are overselling their results. All these statements are not true, only for two out of three outcomes, namely psychological distress and fatigue, but not for perceived stress (see also Figure 3). Hence, I strongly urge the authors to re-write these sentences so that readers are not misled.	1	2	life12060901_makarova	1
+Thank you for pointing out these mistakes, and we trust the editorial office will assist us further with any remaining details in the final stage of the process.	2	1	Besides these two major issues, I detected some smaller errors and miss-spellings (note, this list in not complete). Hence, I have some comments/suggestions that I hope will help the authors to further develop this line of work: Authors’ response:	1	2	life12060901_makarova	1
+Alignment has been fixed.	2	1	Chapter 2 Materials and Methods: Something strange happened to the alignment of heading “2.2 Sample” (page 3, line 128).	1	2	life12060901_makarova	1
+Table 1 has been revised accordingly.	2	1	Please fix this. Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”). The total sample is reported here as 303. However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310). I know, there were 7 missing values within the 310. However, this table is still wrong, it must be 310 and not 303. The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%). Hence, I urge the authors to re-calculate percentages in Table 1. Otherwise, this is misleading the readership.	1	2	life12060901_makarova	1
+Italics have been removed, see revised section 2.3.	2	1	Chapter 2.3 Measures: Sometimes the verbal scales are set in italic (e.g., “0=better than ususal”, page 4, line 173) and sometimes not (e.g., “4=very often”, page 5, line 187).	1	2	life12060901_makarova	1
+We are unsure how this problem occurred. We have fixed the problems in the revised tables.	2	1	Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold. For example, see Table 4 (page 7, line 269): Column Fatgigue, why is ES not printed in bold? Please use a consistent writing style.	1	2	life12060901_makarova	1
+Thank you for pointing out, we have addressed this issue throughout the revised manuscript.	2	1	Please use a consistent writing style throughout the whole manuscript. Results (page 7, line 274): Sometimes fatigue (page 7, line 274) and sometimes with a capital F (Fatigue, page 7, line 283).	1	2	life12060901_makarova	1
+We have corrected the two tables according to this guidance.	4	1	Wrong column headings in tables: I detected that the authors replaced “GHQ” with “psychological distress” in the text (also “PSS” replaced with “perceived stress”). This is correct. Psychological distress is the latent construct which is measured with the instrument GHQ. Unfortunately, the authors missed to replace GHQ by psychological distress in Table 4 (page 7, line 280) and Table 5 (page 7, line 297; see column headings: not “GHQ”, “Fatigue”, and “Perceived stress” but rather “Psychological distress”, “Fatigue”, and “Perceived stress”). Please correct the two tables.	3	2	life12060901_makarova	1
+Thank you for noticing. We have corrected the manuscript according to this guidance.	4	1	"Wrong use of abbreviations: The authors introduced the abbreviation “GHQ-12” for the 12-item General Health Questionnaire (see page 4, line 161), but wrote in the text only ""GHQ"" (e.g., page 5, line 207; page 6, line 256; page 7, line 274). Please replace every single “GHQ” in the text with the correct abbreviation “GHQ-12”. Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e. replace “PSS” with “PSS-10” in the whole text."	3	2	life12060901_makarova	1
+We agree, and we have noted this point in the revised study limitations section.	2	1	The authors admit that, line 367, younger people were overrepresented, but to me, it was that older persons, especially over age 70, who were somehow missed in the survey sampling process. The study offers little to say about long covid among the elderly.	1	2	life12060901_perova	1
+Interesting point, which we have added to the revised Conclusion and Implications section.	2	1	The authors consider a number of factors associated with long COVID and psychological symptoms. I would suggest another factor - frustration with not getting well. Normally, you get sick, you get better (if you don't die). You get injured moderately and you get better with time. Long COVID breaks that pattern - you get sick and stay sick. Like being sick and tired of being sick and tired. Some uncommon illnesses may act that way, but COVID has become endemic and common. What is your common cold lasted for ten months? People would get really irritated and discouraged by such a situation.	1	2	life12060901_perova	1
+Thank you for the kind response.	2	1	I accept the manuscript for its publication in this journal.	1	2	life12060901_perova	1
+The first mentioned statement has been removed altogether in the revised manuscript. The second statement has been modified in accordance with the reviewer’s view (see revised section 4.3).	2	1	The authors said in the statistic section that “statistical significance was set at p<0.05.” (page 5, line 212). However, later they wrote “The descriptively higher proportion of females with long COVID bordered statistical significance (p=0.05)” page 5, line 230) and “However, the different proportions of men and women with long COVID (22% and 33%, respectively) bordered towards statistical significance […]” (page 10, line 318). As a statistician, I felt very mad about this, because the results speak a different language, this is non-significant result.	1	2	life12060901_perova	1
+Thank you for pointing this out. We have removed the relevant section in accordance with this guidance.	2	1	The authors wrote later in the conclusion section “[…] but a non-significant trend was found for the association between female gender and long COVID.” (page 11, line 389). This is a “no-go” in scientific research, the interpretation of “trends” resulted from non-significant results is strongly misleading. Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05). That is the true story. Hence, I urge the authors not to mislead readers and to re-write these sections.	1	2	life12060901_perova	1
+Thank you for pointing this out, we have revised the sentence (see section 4.1).	2	1	The next major issue targets the interpretation of interaction effects. The authors did three models with three different outcomes, namely psychological distress, fatigue, and perceived stress. Authors introduced an interaction effect long COVID x gender into the model in order to test their third hypothesis (“(iii) whether gender moderated the associations between long COVID status and the health outcomes”, page 3, line 119). In the results section, only two (and not three!) interaction effects became significant (psychological distress: p<0.001, fatigue: p<0.05, and perceived stress: p=0.36, see Table 4, page 7, line 269). However, the authors wrote “While participants with long COVID generally perceived more psychological distress, fatigue, and stress than those without long COVID, differences were larger for men than for women” page 9, line 303), Authors’ response:	1	2	life12060901_perova	1
+We have revised the sentence; see section 4.4.	2	1	“[…] men’s perception of poorer health when having long COVID appears to include higher levels of psychological distress, fatigue and perceived stress” (page 11, line 363),	1	2	life12060901_perova	1
+We have revised the sentence; see section 5.	2	1	and finally “Third, long COVID appears to have a stronger effect on men than on women” (page 11, line 401). Again, the authors are overselling their results. All these statements are not true, only for two out of three outcomes, namely psychological distress and fatigue, but not for perceived stress (see also Figure 3). Hence, I strongly urge the authors to re-write these sentences so that readers are not misled.	1	2	life12060901_perova	1
+Alignment has been fixed.	2	1	Chapter 2 Materials and Methods: Something strange happened to the alignment of heading “2.2 Sample” (page 3, line 128).	1	2	life12060901_perova	1
+Table 1 has been revised accordingly.	2	1	Please fix this. Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”). The total sample is reported here as 303. However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310). I know, there were 7 missing values within the 310. However, this table is still wrong, it must be 310 and not 303. The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%). Hence, I urge the authors to re-calculate percentages in Table 1. Otherwise, this is misleading the readership.	1	2	life12060901_perova	1
+Italics have been removed, see revised section 2.3.	2	1	Chapter 2.3 Measures: Sometimes the verbal scales are set in italic (e.g., “0=better than ususal”, page 4, line 173) and sometimes not (e.g., “4=very often”, page 5, line 187).	1	2	life12060901_perova	1
+We are unsure how this problem occurred. We have fixed the problems in the revised tables.	2	1	Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold. For example, see Table 4 (page 7, line 269): Column Fatgigue, why is ES not printed in bold? Please use a consistent writing style.	1	2	life12060901_perova	1
+Thank you for pointing out, we have addressed this issue throughout the revised manuscript.	2	1	Please use a consistent writing style throughout the whole manuscript. Results (page 7, line 274): Sometimes fatigue (page 7, line 274) and sometimes with a capital F (Fatigue, page 7, line 283).	1	2	life12060901_perova	1
+We have corrected the two tables according to this guidance.	4	1	Before a final publication of this paper, I have two minor points left: Wrong column headings in tables: I detected that the authors replaced “GHQ” with “psychological distress” in the text (also “PSS” replaced with “perceived stress”). This is correct. Psychological distress is the latent construct which is measured with the instrument GHQ. Unfortunately, the authors missed to replace GHQ by psychological distress in Table 4 (page 7, line 280) and Table 5 (page 7, line 297; see column headings: not “GHQ”, “Fatigue”, and “Perceived stress” but rather “Psychological distress”, “Fatigue”, and “Perceived stress”). Please correct the two tables.	3	2	life12060901_perova	1
+Thank you for noticing. We have corrected the manuscript according to this guidance.	4	1	"Wrong use of abbreviations: The authors introduced the abbreviation “GHQ-12” for the 12-item General Health Questionnaire (see page 4, line 161), but wrote in the text only ""GHQ"" (e.g., page 5, line 207; page 6, line 256; page 7, line 274). Please replace every single “GHQ” in the text with the correct abbreviation “GHQ-12”. Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e. replace “PSS” with “PSS-10” in the whole text."	3	2	life12060901_perova	1
+In the paper we justify (at some length) the bending modulus we used, which we extrapolated from various sources, and explain why we believe the value in Boal and Ng’s study is likely too low. The reviewer did not question the logic or the sources used to arrive at this value, but nevertheless seemed surprised at the result. The reviewer claims that the parameter falls outside of the physical regime, but doesn’t offer a justification other than that the implied persistence length “seems too large”. However, the large persistence length simply implies that the cyanobacteria are stiff enough that they are largely unaffected by thermal fluctuations. Despite this, the trichomes in our simulations are still quite flexible in practice, as can be seen in Figure 4. Even if the virtual trichomes are somewhat too stiff, we do not feel that this would have a major impact on the results.	2	1	The literature seems to indicate a low value of the bending modulus (alpha) that corresponds to a persistence length of 500 microns. The authors instead use alpha = 2 × 10−21 Nm2, which is 1000× larger. With this value, the persistence length = alpha/(kbT) = 0.5 metres. This seems orders of magnitude too large, especially in the face of the results of Boal and Ng [44]. The authors need to be in a physical regime of parameter space in order to confront their results with observed patterns. Otherwise, they are just fitting a non-linear phenomenon with a (potentially inappropriate) non-linear model— in this case agreement does not provide firm insight into the physical system.	1	2	life4030433_makarova	1
+We added the following statement to the end of Section 3.3: “The simulation results of the previous section for β > 0.5 are unlikely to be affected by the domain size, however, since the features in those simulations are on a much smaller scale than the domain size and are also more chaotic, as seen in the correlation length and the global alignment (Figure 5a,d).”	2	1	In Section 3.3 the authors describe how an increase in system size indicated that one of the results presented earlier in the paper (at beta = 0.5) was an artifact of a smaller system size. This brings into question all results at the smaller system size, potentially including all the quantitative results of the paper. The authors should be able to argue that the other results will not depend on system size. The correlation length in Figure 3b is by eye more than the system size, indicating a potential qualitative change with increasing system size (which the authors find). The correlation length in other figures is less than the system size, indicating that they may be fine. The authors need to add some of this discussion to reassure the reader.	1	2	life4030433_makarova	1
+We added the following clarification to the end of the non-dimensionalization section: “In the subsequent sections, simulations for β = 0 use a modified interaction force such that there is no cohesion, but the hard core repulsion is maintained, i.e., 0 0 c F = for 0 h ≥ , otherwise 0 c c F F = with β = 0.125.” (4)	2	1	The authors find that results at beta = 0 are most similar to the experimental reticulate patterns. In the force model, there are three forces: elastic, gliding, and contact/cohesion. The contact/cohesion force is presented in equation 12, and is proportional to epsilon. After Equation 23, the authors define beta = epsilon/(zeta⋅v0), implying that there will be no contact/cohesion forces when beta = 0. However, in the results of Figures 3, 4, and 5 the results for beta = 0 clearly show interactions between the filaments. The fourth paragraph of section 2.9 describes a hard-core interaction, which appears to be the source of the patterns when beta = 0. This hard-core interaction should be mentioned after Equation 12, since it is not simply an implementation issue but the dominant interaction with beta = 0.	1	2	life4030433_makarova	1
+We added two references to justify using the Lennard–Jones potential.	2	1	All the results depend on the cohesion forces, which are described by a Lennard–Jones function in Equation 12, in Section 2.5. However, this way of approximating cohesion is not well justified, and in Section 2.9 (several sections later) the authors mention that in reality the attractive force “does not really exist” and that the trichomes cohere after contact. The authors are using an LJ interaction to approximate contact adhesion. They should say why, and address how good of an approximation they expect this to be.	1	2	life4030433_makarova	1
+We added the following clarification: “This is simulated by generating a pseudo-random number x following a uniform distribution X ~ U(0,1), and reversing the gliding direction if x < ω⋅∆t.	2	1	In Section 2.4, it is stated that a stochastic process determines P, but no details are given. There should be some description, if brief, for this process. What distribution is used?	1	2	life4030433_makarova	1
+We amended the following sentences of the discussion: “These differences may be due to the fact that we use a shallow domain (7.5 microns) to reduce the simulation run time, which may cause the pattern to be “squashed” as the domain ceiling prevents ridges from growing vertically” “Restricting the virtual trichomes such that they may only glide when in contact with the substratum or another trichome may promote further aggregation of the trichomes from streams into ridges, as a trichome would be less likely to successfully break away from a stream because it would lose much of its propulsive force as it lost contact with neighboring trichomes and/or the substratum.”	2	1	The authors mention in the discussion that they may get stronger results for a deeper system, or for trichomes that may only glide freely near a surface or other trichome. In terms of the understanding gained from the modelling/simulations, how would these lead to qualitative changes in the observed patterns?	1	2	life4030433_makarova	1
+We added the following sentence to Section 2.4: “Gliding requires contact with some (semi-)solid substrate in order to provide a reaction force to the gliding mechanism. We assume that the trichomes are immersed in highly viscous medium that allows them to glide freely in all directions. This medium could consist of the EPS the trichomes produce copiusly when gliding [Hoiczyk2000].” (8)	2	1	The viscosity mu = 1 Pa s is used. This value is approximately 1000 times higher than the viscosity of water, which is the environment of filamentous cyanobacteria. A reference is given, but there should also be a brief justification in the text.	1	2	life4030433_makarova	1
+The exact force is now given.	2	1	The authors do not include a description of the reaction force for the top and bottom planes. A brief explanation would be appropriate.	1	2	life4030433_makarova	1
+The bibliography now has volume and page numbers.	2	1	None of the references at the end of the paper contain journal volume or page numbers. They should.	1	2	life4030433_makarova	1
+The reviewer did not provide any examples where he/she felt a reference was warranted. We cited 55 sources, which we feel offers reasonable background and justification for our assumptions.	2	1	In general, and especially in the introduction, many statements are made without accompanying references.	1	2	life4030433_makarova	1
+References to the movies were added to the results section.	2	1	There are supplemental movies attached. These are not referred to in the text.	1	2	life4030433_makarova	1
+We added the following sentence to the end of Section 2.8: “This value corresponds to a persistence length of 0.49 m, which implies that the trichomes would be practically unaffected by thermal fluctuations.”	4	1	At the end of the discussion of the bending modulus (in Section 2.8, on page 11), the implied persistence length and the irrelevance of thermal fluctuations should be explicitly mentioned.	3	2	life4030433_makarova	1
+We added the following paragraph to the discussion: “Many of the model parameters, such as trichome length, diameter, gliding speed, reversal frequency, etc. are easily measured and are well known. However, the bending modulus is more difficult to measure directly. We attempted to use the bending modulus implied from the relation α= kBT ε and Boal and Ng’s (2010, p. 4625) measurements of the trichome persistence length, but the resulting value seemed too low compared to measurements of other bacteria and in practice the virtual trichomes appeared flaccid during simulations. It is possible that the flexure seen in Boal and Ng’s trichomes was due more to the motility of the trichomes than random thermal fluctuations, in which case the above relation would no longer be valid, and a more complex model would be required to associate the observed geometry of the trichomes to their bending modulus. For example, Wolgemuth (2005) used an elastic model to estimate the bending modulus of M. xanthus by fitting the model to the flailing motions of a Myxobacterium stuck at one end.”	4	1	In Section 4, the discussion section of the paper, the topic of the bending modulus is returned to for a paragraph. This paragraph needs to explicitly address why the measurements of Boal and Ng were not used, i.e., why Boal and Ng misinterpreted their data. In particular the authors should suggest how the measurements are significant measurements to not use. In particular this paragraph needs to R8 suggest how the measurement should be done to properly test their suggested values for the bending modulus, and so resolve the disagreement with Boal and Ng.	3	2	life4030433_makarova	1
+The symbol used to denote trichome length was changed to an uppercase lambda. The , ⋅⋅ is often used to denote a tuple in computer science, in this case a pair. The superscripts should have been subscripts. The notation used in Equations (5)–(8) was adopted from Bergou et al’s paper. h-hat is in fact a unit vector parallel to h, and we’ve clarified the text to this point.	2	1	• ˆ h looks like a unit vector parallel to h. Is there a reason that h only has a ˆ ⋅ in Equation (12)?	1	2	life4030433_makarova	1
+We added a few sentences to the introduction that expand a bit on similar structures in the Petroff and Walter references.	2	1	Walter (ref 7) describes the formation of “clumps” which are very similar to the structures describes by Shepard. Shepard and Sumner provide better images (Walter only shows sketches), but it might be worth mentioning that the phenomenon is more general. As I recall, Petroff et al. (ref 13) show similar patterns.	1	2	life4030433_makarova	1
+Actually, as Reviewer 1 noted, we use a viscosity that is 1000 times greater than that of water (following Wolgemuth et al. 2005), for the very reasons the reviewer mentioned.	2	1	In the force balance, the drag coeffcient is assumed to be the viscosity of water. Since the cells glide over a surface and next to other cells, it would seem that cell–cell friction and cell–surface friction might be at least as important as hydrodynamic drag. What about the viscosity of the material used in the “slime jet”?	1	2	life4030433_makarova	1
+The statement has been clarified and is now: “The sum of the interaction’s opposing forces and torques is zero, thereby ensuring Newton's third law is respected.”	2	1	Following Equation (13), it is stated that “The interpolation ensures the net force and net torque of the interaction forces are null”. This confuses me so I suspect that I have misunderstood something. In an overdamped system, such as this one, all forces and torques are balanced by drag on the cell. Is this all that is happening in this derivation? The text reads like certain forces and torques are assumed to vanish. Are these forces and torques introduced by the interpolation and the parameterization is chosen to cancel artifacts? Or, are there internal forces and and torques acting on the vertices that cancel as a result of Newton’s second law? Please clarify this point.	1	2	life4030433_makarova	1
+Our estimate is simplistic, however we were unable to find a reliable measurement of this parameter in the literature, and so we resorted to some simple extrapolations.	2	1	In the estimation of the Young’s modulus it is assumed that the bending resistance is due to the cell wall. I suspect that this is a very poor approximation, accounting for the two order of magnitude difference between the estimated and benchmark values of Y. It would seem that the resistance is controlled or at least influenced by additional factors such as protein expression (notably MreB) and osmotic pressure. If my intuition is wrong (I am not an expert on cell morphology), I would appreciate knowing why.	1	2	life4030433_makarova	1
+There are indeed additional parameters. We have added a complete list to Section 2.8. Many are explored to some extent in the results (beta, reversal frequency, domain dimensions, trichome R7 density), whereas most of the physical characteristics of the trichomes are known from published sources (diameter, length, gliding speed, bending modulus), and finally N was simply chosen so that trichomes had a “smooth” appearance.	2	1	If I understand correctly, there are six important length scales in the problem, the size of the domain DH , the spacing between trichomes 13 ρ − , 0 , , L l αν ζ = Θ, and the trichome length L = Nl (also denoted L in the manuscript), giving five dimensionless numbers. However, in the non-dimensionalization, the authors state that there are only two parameters: ρ and Nl γ θ = . Unless I am mistaken, there are additional parameters like l/L, Nl/D, and N that the authors hold fixed but may influence the details of the numerics.	1	2	life4030433_makarova	1
+To avoid confusion on this point we have replaced this statement with the following: “We find that these errors are infrequent and acceptable as a trade-off for increased computational performance.” We have also added a comment to Section 2.5 regarding the Lennard–Jones interaction: “The attractive force has a short range, and is practically zero at h = 2θ.”	2	1	It seems a little odd to posit an attractive force in the derivation (Equation (12)) and then ignore/dismiss it in the numerics (“… in reality the attractive force does not really exist, trichomes only cohere after they have come into contact” page 12). Clearly, this is done to (quite reasonably) avoid modeling steric interactions with hard bodies. It was a little distracting to think that sticky adhesion was decaying like 6 ( ) h Θ . Perhaps note after Equation (12) that the attractive force decreases to zero at a finite distance in the numerics.	1	2	life4030433_makarova	1
+It would be interesting to explore the approach the reviewer suggested; we have not yet explored more coarse grained models.	2	1	Can you coarse grain this model to make it analytically tractable? The basic phenomena suggest something like an active shear-thinning uid? This would make it easier to understand the bifurcations in the dynamics and may require fewer parameters. Just something to think about.	1	2	life4030433_makarova	1
+Shepard et al. claim that the formation of the reticulate pattern occurs on a faster scale than cell growth and division, and so we did not consider this in our model. Including this might make the results more robust, however, and we added a comment to the discussion.	2	1	Notably, the introduction of reproduction does not introduce another parameter. As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+). Thus, the length of a filament is not a model parameter. Rather, length is determined dynamically as ~ N k τ + (10)	1	2	life4030433_makarova	1
+We had plans to include a quantitative comparison, however we were not able obtain a good dataset for doing so. It would be interesting to replicate Shepard’s experiments taking care to collect enough good quality images/movies for a quantitative analysis on the patterns.	2	1	This project would benefit immeasurably from a quantitative comparison to experiments. I understand that this is beyond the scope of this paper, but I strongly encourage future work in this direction.	1	2	life4030433_makarova	1
+Replicating Shepard and Sumner’s results was our primary objective for this paper, as we state in the discussion. In future work we would like to increase the scale of the simulations by adding more trichomes and a deeper domain to (hopefully) get more robust reticulate formation. We would also like to do a quantitative analysis on experimental data to in order to fit the model to the data. Finally, we would like to see whether this same minimal system plus photomovement is sufficient to produce the cone-shaped structures documented by Walter, Petroff and others. If successful, we could then link macroscopic features of similar stromatolites back to the parameters of the trichomes, as well as understand the contribution of each factor in the model to building these structures. We have added this explanation to the conclusions section.	2	1	In the introduction, this project is put into the context of stromatolites. In the discussion/conclusion, I would appreciate to hear the authors thoughts on what this model teaches us about stromatolites. In the current form I am left to conclude only that communities of filamentous bacteria can develop reticulated patterns similar to forms observed in the fossil record. This conclusion does not offer much more than Shepard’s experimental observation of this result. Can we use the details of this model to, for example, estimate the cohesion β of ancient cells? Do these results provide any insight into the identity of stromatolite building cells. As far as I can tell, nothing in this model suggests that photosynthesis (much less oxygenic photosynthesis) is required to form these patterns. It seems that this model shows that polygonal patters are a generic feature of elongated gliding cells and is independent of metabolism. A quick google search produced this video of mat of the sulfur-oxidizing bacteria Beggiatoa spp. http://vimeo.com/57205513 (note: I have no idea of the details of these observations so any similarity may be spurious).	1	2	life4030433_makarova	1
+We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low. The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display. In the paper we justify (at some length) the bending modulus we used, which we extrapolated from various sources, and explain why we believe the value in Boal and Ng’s study is likely too low. The reviewer did not question the logic or the sources used to arrive at this value, but nevertheless seemed surprised at the result. The reviewer claims that the parameter falls outside of the physical regime, but doesn’t offer a justification other than that the implied persistence length “seems too large”. However, the large persistence length simply implies that the cyanobacteria are stiff enough that they are largely unaffected by thermal fluctuations. Despite this, the trichomes in our simulations are still quite flexible in practice, as can be seen in Figure 4. Even if the virtual trichomes are somewhat too stiff, we do not feel that this would have a major impact on the results.	2	1	The literature seems to indicate a low value of the bending modulus (alpha) that corresponds to a persistence length of 500 microns. The authors instead use alpha = 2 × 10−21 Nm2, which is 1000× larger. With this value, the persistence length = alpha/(kbT) = 0.5 metres. This seems orders of magnitude too large, especially in the face of the results of Boal and Ng [44]. The authors need to be in a physical regime of parameter space in order to confront their results with observed patterns. Otherwise, they are just fitting a non-linear phenomenon with a (potentially inappropriate) non-linear model— in this case agreement does not provide firm insight into the physical system.	1	2	life4030433_perova	1
+We added the following statement to the end of Section 3.3: “The simulation results of the previous section for β > 0.5 are unlikely to be affected by the domain size, however, since the features in those simulations are on a much smaller scale than the domain size and are also more chaotic, as seen in the correlation length and the global alignment (Figure 5a,d).”	2	1	In Section 3.3 the authors describe how an increase in system size indicated that one of the results presented earlier in the paper (at beta = 0.5) was an artifact of a smaller system size. This brings into question all results at the smaller system size, potentially including all the quantitative results of the paper. The authors should be able to argue that the other results will not depend on system size. The correlation length in Figure 3b is by eye more than the system size, indicating a potential qualitative change with increasing system size (which the authors find). The correlation length in other figures is less than the system size, indicating that they may be fine. The authors need to add some of this discussion to reassure the reader.	1	2	life4030433_perova	1
+We added the following clarification to the end of the non-dimensionalization section: “In the subsequent sections, simulations for β = 0 use a modified interaction force such that there is no cohesion, but the hard core repulsion is maintained, i.e., 0 0 c F = for 0 h ≥ , otherwise 0 c c F F = with β = 0.125.”	2	1	The authors find that results at beta = 0 are most similar to the experimental reticulate patterns. In the force model, there are three forces: elastic, gliding, and contact/cohesion. The contact/cohesion force is presented in equation 12, and is proportional to epsilon. After Equation 23, the authors define beta = epsilon/(zeta⋅v0), implying that there will be no contact/cohesion forces when beta = 0. However, in the results of Figures 3, 4, and 5 the results for beta = 0 clearly show interactions between the filaments. The fourth paragraph of section 2.9 describes a hard-core interaction, which appears to be the source of the patterns when beta = 0. This hard-core interaction should be mentioned after Equation 12, since it is not simply an implementation issue but the dominant interaction with beta = 0.	1	2	life4030433_perova	1
+We added two references to justify using the Lennard–Jones potential.	2	1	All the results depend on the cohesion forces, which are described by a Lennard–Jones function in Equation 12, in Section 2.5. However, this way of approximating cohesion is not well justified, and in Section 2.9 (several sections later) the authors mention that in reality the attractive force “does not really exist” and that the trichomes cohere after contact. The authors are using an LJ interaction to approximate contact adhesion. They should say why, and address how good of an approximation they expect this to be.	1	2	life4030433_perova	1
+We added the following clarification: “This is simulated by generating a pseudo-random number x following a uniform distribution X ~ U(0,1), and reversing the gliding direction if x < ω⋅∆t.	2	1	In Section 2.4, it is stated that a stochastic process determines P, but no details are given. There should be some description, if brief, for this process. What distribution is used?	1	2	life4030433_perova	1
+We amended the following sentences of the discussion: “These differences may be due to the fact that we use a shallow domain (7.5 microns) to reduce the simulation run time, which may cause the pattern to be “squashed” as the domain ceiling prevents ridges from growing vertically” “Restricting the virtual trichomes such that they may only glide when in contact with the substratum or another trichome may promote further aggregation of the trichomes from streams into ridges, as a trichome would be less likely to successfully break away from a stream because it would lose much of its propulsive force as it lost contact with neighboring trichomes and/or the substratum.”	2	1	The authors mention in the discussion that they may get stronger results for a deeper system, or for trichomes that may only glide freely near a surface or other trichome. In terms of the understanding gained from the modelling/simulations, how would these lead to qualitative changes in the observed patterns?	1	2	life4030433_perova	1
+We added the following sentence to Section 2.4: “Gliding requires contact with some (semi-)solid substrate in order to provide a reaction force to the gliding mechanism. We assume that the trichomes are immersed in highly viscous medium that allows them to glide freely in all directions. This medium could consist of the EPS the trichomes produce copiusly when gliding [Hoiczyk2000].” (8)	2	1	The viscosity mu = 1 Pa s is used. This value is approximately 1000 times higher than the viscosity of water, which is the environment of filamentous cyanobacteria. A reference is given, but there should also be a brief justification in the text.	1	2	life4030433_perova	1
+The exact force is now given.	2	1	The authors do not include a description of the reaction force for the top and bottom planes. A brief explanation would be appropriate.	1	2	life4030433_perova	1
+The bibliography now has volume and page numbers.	2	1	None of the references at the end of the paper contain journal volume or page numbers. They should.	1	2	life4030433_perova	1
+The reviewer did not provide any examples where he/she felt a reference was warranted. We cited 55 sources, which we feel offers reasonable background and justification for our assumptions.	2	1	In general, and especially in the introduction, many statements are made without accompanying references.	1	2	life4030433_perova	1
+References to the movies were added to the results section.	2	1	There are supplemental movies attached. These are not referred to in the text.	1	2	life4030433_perova	1
+We added the following sentence to the end of Section 2.8: “This value corresponds to a persistence length of 0.49 m, which implies that the trichomes would be practically unaffected by thermal fluctuations.”	4	1	At the end of the discussion of the bending modulus (in Section 2.8, on page 11), the implied persistence length and the irrelevance of thermal fluctuations should be explicitly mentioned.	3	2	life4030433_perova	1
+We added the following paragraph to the discussion: “Many of the model parameters, such as trichome length, diameter, gliding speed, reversal frequency, etc. are easily measured and are well known. However, the bending modulus is more difficult to measure directly. We attempted to use the bending modulus implied from the relation α= kBT ε and Boal and Ng’s (2010, p. 4625) measurements of the trichome persistence length, but the resulting value seemed too low compared to measurements of other bacteria and in practice the virtual trichomes appeared flaccid during simulations. It is possible that the flexure seen in Boal and Ng’s trichomes was due more to the motility of the trichomes than random thermal fluctuations, in which case the above relation would no longer be valid, and a more complex model would be required to associate the observed geometry of the trichomes to their bending modulus. For example, Wolgemuth (2005) used an elastic model to estimate the bending modulus of M. xanthus by fitting the model to the flailing motions of a Myxobacterium stuck at one end.”	4	1	In Section 4, the discussion section of the paper, the topic of the bending modulus is returned to for a paragraph. This paragraph needs to explicitly address why the measurements of Boal and Ng were not used, i.e., why Boal and Ng misinterpreted their data. In particular the authors should suggest how the measurements are significant measurements to not use. In particular this paragraph needs to R8 suggest how the measurement should be done to properly test their suggested values for the bending modulus, and so resolve the disagreement with Boal and Ng.	3	2	life4030433_perova	1
+The symbol used to denote trichome length was changed to an uppercase lambda. The , ⋅⋅ is often used to denote a tuple in computer science, in this case a pair. The superscripts should have been subscripts. The notation used in Equations (5)–(8) was adopted from Bergou et al’s paper. h-hat is in fact a unit vector parallel to h, and we’ve clarified the text to this point.	2	1	Notation: • L is used both for the length of the trichome Nl and in the non-dimensionalization as 0 αν ζ . • I have never seen , ⋅⋅used to define an edge. It looks like an inner product. Is this standard? • In Equations (5)–(8), is there any difference between subscripts and superscripts? • It is confusing that b κ is a vector that is not simply related to b. I initially thought that κ was a new parameter. Similarly, it is odd that e is a vector and [e] is a matrix. Why not use the Levi–Civita tensor? • ˆ h looks like a unit vector parallel to h. Is there a reason that h only has a ˆ ⋅ in Equation (12)?	1	2	life4030433_perova	1
+We added a few sentences to the introduction that expand a bit on similar structures in the Petroff and Walter references.	2	1	Walter (ref 7) describes the formation of “clumps” which are very similar to the structures describes by Shepard. Shepard and Sumner provide better images (Walter only shows sketches), but it might be worth mentioning that the phenomenon is more general. As I recall, Petroff et al. (ref 13) show similar patterns.	1	2	life4030433_perova	1
+Actually, as Reviewer 1 noted, we use a viscosity that is 1000 times greater than that of water (following Wolgemuth et al. 2005), for the very reasons the reviewer mentioned.	2	1	In the force balance, the drag coeffcient is assumed to be the viscosity of water. Since the cells glide over a surface and next to other cells, it would seem that cell–cell friction and cell–surface friction might be at least as important as hydrodynamic drag. What about the viscosity of the material used in the “slime jet”?	1	2	life4030433_perova	1
+The statement has been clarified and is now: “The sum of the interaction’s opposing forces and torques is zero, thereby ensuring Newton's third law is respected.”	2	1	Following Equation (13), it is stated that “The interpolation ensures the net force and net torque of the interaction forces are null”. This confuses me so I suspect that I have misunderstood something. In an overdamped system, such as this one, all forces and torques are balanced by drag on the cell. Is this all that is happening in this derivation? The text reads like certain forces and torques are assumed to vanish. Are these forces and torques introduced by the interpolation and the parameterization is chosen to cancel artifacts? Or, are there internal forces and and torques acting on the vertices that cancel as a result of Newton’s second law? Please clarify this point.	1	2	life4030433_perova	1
+Our estimate is simplistic, however we were unable to find a reliable measurement of this parameter in the literature, and so we resorted to some simple extrapolations.	2	1	In the estimation of the Young’s modulus it is assumed that the bending resistance is due to the cell wall. I suspect that this is a very poor approximation, accounting for the two order of magnitude difference between the estimated and benchmark values of Y. It would seem that the resistance is controlled or at least influenced by additional factors such as protein expression (notably MreB) and osmotic pressure. If my intuition is wrong (I am not an expert on cell morphology), I would appreciate knowing why.	1	2	life4030433_perova	1
+There are indeed additional parameters. We have added a complete list to Section 2.8. Many are explored to some extent in the results (beta, reversal frequency, domain dimensions, trichome R7 density), whereas most of the physical characteristics of the trichomes are known from published sources (diameter, length, gliding speed, bending modulus), and finally N was simply chosen so that trichomes had a “smooth” appearance.	2	1	If I understand correctly, there are six important length scales in the problem, the size of the domain DH , the spacing between trichomes 13 ρ − , 0 , , L l αν ζ = Θ, and the trichome length L = Nl (also denoted L in the manuscript), giving five dimensionless numbers. However, in the non-dimensionalization, the authors state that there are only two parameters: ρ and Nl γ θ = . Unless I am mistaken, there are additional parameters like l/L, Nl/D, and N that the authors hold fixed but may influence the details of the numerics.	1	2	life4030433_perova	1
+To avoid confusion on this point we have replaced this statement with the following: “We find that these errors are infrequent and acceptable as a trade-off for increased computational performance.” We have also added a comment to Section 2.5 regarding the Lennard–Jones interaction: “The attractive force has a short range, and is practically zero at h = 2θ.”	2	1	It seems a little odd to posit an attractive force in the derivation (Equation (12)) and then ignore/dismiss it in the numerics (“… in reality the attractive force does not really exist, trichomes only cohere after they have come into contact” page 12). Clearly, this is done to (quite reasonably) avoid modeling steric interactions with hard bodies. It was a little distracting to think that sticky adhesion was decaying like 6 ( ) h Θ . Perhaps note after Equation (12) that the attractive force decreases to zero at a finite distance in the numerics.	1	2	life4030433_perova	1
+It would be interesting to explore the approach the reviewer suggested; we have not yet explored more coarse grained models.	2	1	The parameter space is rather large, and only a small subspace is explored in the numerics. This is fine, but it leaves a lot of interesting questions un-addressed. For example, is the transition from broad streams (Figure 4a) to loops (Figure 4d) continuous or are there bifurcations/phase transitions? Can you coarse grain this model to make it analytically tractable? The basic phenomena suggest something like an active shear-thinning uid? This would make it easier to understand the bifurcations in the dynamics and may require fewer parameters. Just something to think about.	1	2	life4030433_perova	1
+Shepard et al. claim that the formation of the reticulate pattern occurs on a faster scale than cell growth and division, and so we did not consider this in our model. Including this might make the results more robust, however, and we added a comment to the discussion.	2	1	In this model, the density of cells is assumed to be constant. However, the timescale of the simulations is comparable to the doubling time of many bacteria. Because cell density is found to R6 be important in determining the morphology of communities, I suspect that reproduction is important in understanding the patterns. This point might be worth addressing in the text.	1	2	life4030433_perova	1
+We had plans to include a quantitative comparison, however we were not able obtain a good dataset for doing so. It would be interesting to replicate Shepard’s experiments taking care to collect enough good quality images/movies for a quantitative analysis on the patterns.	2	1	This project would benefit immeasurably from a quantitative comparison to experiments. I understand that this is beyond the scope of this paper, but I strongly encourage future work in this direction.	1	2	life4030433_perova	1
+Replicating Shepard and Sumner’s results was our primary objective for this paper, as we state in the discussion. In future work we would like to increase the scale of the simulations by adding more trichomes and a deeper domain to (hopefully) get more robust reticulate formation. We would also like to do a quantitative analysis on experimental data to in order to fit the model to the data. Finally, we would like to see whether this same minimal system plus photomovement is sufficient to produce the cone-shaped structures documented by Walter, Petroff and others. If successful, we could then link macroscopic features of similar stromatolites back to the parameters of the trichomes, as well as understand the contribution of each factor in the model to building these structures. We have added this explanation to the conclusions section.	2	1	In the introduction, this project is put into the context of stromatolites. In the discussion/conclusion, I would appreciate to hear the authors thoughts on what this model teaches us about stromatolites. In the current form I am left to conclude only that communities of filamentous bacteria can develop reticulated patterns similar to forms observed in the fossil record. This conclusion does not offer much more than Shepard’s experimental observation of this result. Can we use the details of this model to, for example, estimate the cohesion β of ancient cells? Do these results provide any insight into the identity of stromatolite building cells. As far as I can tell, nothing in this model suggests that photosynthesis (much less oxygenic photosynthesis) is required to form these patterns. It seems that this model shows that polygonal patters are a generic feature of elongated gliding cells and is independent of metabolism. A quick google search produced this video of mat of the sulfur-oxidizing bacteria Beggiatoa spp. http://vimeo.com/57205513 (note: I have no idea of the details of these observations so any similarity may be spurious).	1	2	life4030433_perova	1
+We are not sure if type IV pili are visible in our EM figures. They are very thin (less than 1 nm) and normally visualized by negative staining. In the published EM figures of Phormidium (conventional and cryo EM), type IV pili have not been noted. The filaments that we see in our EM figures could be components of sheath materials.	2	1	As depicted in some of the electron micrographs in Figure 3, the authors describe the presence of thin filaments within the sheaths surrounding the motile filaments. Is it possible that these are in fact type IV pili? How does the structure compare to other published reports of type IV pili?	1	2	life4040819_makarova	1
+This was corrected.	2	1	In Section 3.5, Analysis of Movement of Filaments, the descriptions were confusing at times. For instance Line 253 states “only in some exceptional cases (the straight line in Figure 6D), net displacement was achieved”. Net displacement presumably refers to any final position after a cycle that differs from the starting position. In other words, net displacement would refer to any value other than zero in Figure 6D. All of the points on this graph appear to indicate net displacement, albeit some less than others. Do the authors mean to state that only in a few instances was a large net displacement achieved? Otherwise the authors’ statement does not appear correct. Perhaps using numerical values instead of statements such as “the average velocity was quite limited” would help to clarify.	1	2	life4040819_makarova	1
+We did not find such an indication in the literature. This point was added in the text.	2	1	The authors observe that the velocity of movement was maximal immediately after a reversal and then declined exponentially until the next reversal. The authors then suggest that this observation is theoretically inconsistent with type IV pili or focal adhesion complexes as the driving force for movement. Are there any available published reports quantifying the motility of organisms using these systems that would corroborate this empirically? In short, has it been demonstrated that bacteria moving by twitching motility or focal adhesion do not show a similar pattern for velocity of movement?	1	2	life4040819_makarova	1
+The expression was modified.	2	1	The authors state that supracellular structures emerge from unordered motion and are therefore not encoded for genetically. While their model for spiral formation does not require a genetic program for controlling motility, it may not always be the case for other supercellular structures. In other words, it may not always be true that supercellular structures are not genetically determined.	1	2	life4040819_makarova	1
+Line 45—This was corrected.	2	1	Line 45—suggest adding “to” before “back-and-forth motion...”	1	2	life4040819_makarova	1
+Lines 268 and 269—These were corrected. We found many similar cases, too.	2	1	Lines 268 and 269—“(D)” and “(E)” should be bolded to be consistent	1	2	life4040819_makarova	1
+Line 287—Two values were added.	2	1	Line 287—appears to be missing e-values. The beginning of this sentence indicates 10 genes and then only gives 8 respective e-values for the 10 genes.	1	2	life4040819_makarova	1
+Line 290—Some description was added.	2	1	Line 290—do orf303 and orf297 have homology to any characterized proteins? Are they conserved hypotheticals? Is there any indication of the function for these?	1	2	life4040819_makarova	1
+Line 415—This was corrected.	2	1	Line 415—“single” appears to be formatted differently than the rest of the text	1	2	life4040819_makarova	1
+The choice of EM technique might have been wrong in visualizing the so-called oscillin fibrils, but was effective in visualizing membrane structures. The absence of “oscillin” was based on the genomic analysis but not on EM images. In fact, “oscillin” is a glycine-rich large protein which is poorly conserved in bacteria. In the reported case of P. uncinatum, oscillin might be important in forming fibrils, but in other organisms, other proteins could function as surface fibrils aligning the flow of slime. We agree that the method of fixation was not good for preserving surface structures such as oscillin fibrils, if present.	2	1	About 20 years ago it was shown that in order to preserve crucial ultrastructural features of cyanobacterial cell walls, the cells have to be processed using cryo-procedures (Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395). The authors use an outdated and very artifact-laden method to process their samples. This choice is responsible for a number of shortcomings that need to be addressed: The lack of certain features such as the S-layer and the oscillin fibrils in the reported pictures are attributable to the choice of preparation, as they are clearly present in two species of the same genus described in the above cited paper. Moreover, these structures would explain the rotation of the cells that now is somewhat “mysteriously” explained by a helical slime ejection that is not supported by the observed arrangement of the nozzles. Therefore the authors should do the following: Response:	1	2	life4040819_makarova	1
+This point was added in Section 3.7, in which oscillin is discussed. In general, pore-inclination and oscillin fibrils are not exclusive.	2	1	(a) Clearly spell out in the text why there are no oscillin fibrils and surface structures visible and compare their results with the published results of the cell wall structure of the species of the same genus prepared using cryo-procedures.	1	2	life4040819_makarova	1
+The quality of Figure 1A was rather bad. The contrast was corrected and a new figure was inserted. In the new Figure 1A, the sheath is quite visible. The quality of EM figure is very low as embedded in an MS word file. We provide a better quality figure as supplemental Figure S1. The use of “sheath” is therefore correct in the paragraph describing the figure. In the model, the word “sliding” was not understood in the sense in which we wanted it to be. This is lateral sliding that provokes curvature of the filament. This point was clarified in the text.	2	1	(b) There is no clear distinction in the text between the “sheath” and the secreted “slime” of the cells, which are two completely different structures both physically and chemically (Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Figure 1A shows a filament that lacks the “sheath”, while in other pictures the structure is visible. The reason for this is the following: cyanobacteria of the genus Phormidium build over time a carbohydrate layer on their surface that is physically attached to their cell surface, usually called the sheath. Filaments that are ensheathed are non-motile! In contrast, the slime that is secreted by gliding filaments of the same species is not visible in TEM preparations, even when cryo-preservation methods are used (see Figure 1A of the ms and Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395 and Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Therefore, the authors should go through their ms and make a careful distinction between these two structures. For example, in the discussion it sounds as if the “sheath” is preventing the R5 cells from sliding, however this is the slime tube that is secreted that is not physically attached to the cell surface at all.	1	2	life4040819_makarova	1
+What is “the basal body part”? Basal body is an organelle of eukaryotic cell. If this means the connection of junctional pore tube and inner membrane, it is clearly seen in the figure, as well as in old paper by Halfen and Castenholz. A high quality figure is provided as Supplementary Figure S1. The “bulges” in the inner membrane are also visible. In the Line 183, the description was revised.	2	1	(c) Please omit the description of the basal body part of the junctional pores or provide substantially better pictures. Even after careful inspection of your images, I cannot convince myself of seeing these parts. Moreover, in cryo-preserved and freeze-fractured cells there are no such structures visible. That does not mean that they don’t exist, it may only mean that none of the so far used preservation methods reveals these structures.	1	2	life4040819_makarova	1
+In Figure 3D, the junctional pores are seen as an array of tilted tubes. The mechanism of rotation is still a mystery because oscillin itself is not present in the strain KS. This is discussed in Section 3.7 with reservations on our hypothesis of inclined junctional pores. On the other hand, the involvement of fibrils in the rotation and locomotion was proposed a long time ago, whether the fibrils are made of oscillin or other proteins. In this respect, we still have to work a lot to identify the real mechanism of rotation.	2	1	(d) Please omit the description about helical tilted junctional pores. These structures are clearly not arranged in this way and the reasons for rotation are the unfortunately not-preserved surface structures mentioned above.	1	2	life4040819_makarova	1
+This is not a good way of naming headings. A heading should not be a phrase or conclusion, but should be descriptive words.	2	1	Results section headings and figure captions should be phrased as conclusions, supported by the data presented, when appropriate (can be a minor change, for example 3.2 could be titled, “Phormidium colonies form divergent structures,” or larger, for example 3.5 could be titled, “Individual filaments undergo directional reversals, with maximum velocity immediately following a reversal.”	1	2	life4040819_makarova	1
+This was described in Section 3.7.	2	1	In the discussion there are further points that need to be addressed: (a) Please clarify the source of the helical flow of the slime, which is most likely due to the presence of the not-preserved helically arranged cell surface proteins not the arrangement or tilt of the junctional pores.	1	2	life4040819_makarova	1
+This point was corrected.	2	1	(b) In the model that describes the potential activity of the junctional pores (How myxobacteria glide. Curr. Biol. 2002, 12, 369–377) it is not suggested that the nozzle actually “contracts” as mentioned in the ms. The swelling of the slime material in the nozzle fills the nozzle and eventually will generate a counter-force of the nozzle walls of the nozzle that results in the ejection of the slime. Please rephrase the text accordingly.	1	2	life4040819_makarova	1
+There might be a misunderstanding in the first part. The rotation of filament does not drive macroscopic rotation or spiral formation. What we describe in the text was the switch to turn to the left was governed by the filament rotation, but the real formation of a spiral is driven by the locomotion of the filament. The cited paper described the clumping of Anabaena cylindria in a dense culture. The clumping or aggregation in Arthrospira (Ohmori group) is mediated by cAMP, but A. cylindrica might be different. I have been using Anabaena for about 40 years, but I have never seen Anabaena filaments form a spiral on agar plates. Spiral formation and clumping are different phenomena. It is difficult to discuss the relationship (if any) between clumping and rotation.	2	1	(c) When the cells form spirals, there are two components to this process: the formation of spirals and their macroscopic sense of rotation: clock- or counterclockwise. In the discussion it is assumed that the rotation of the filaments is crucial for both of these components. If this would be true, then no spirals should be observed in non-rotating filamentous gliding cyanobacteria. However, this is not the case. Anabaena spec. a non-rotating species can generate spirals (see Figure X; Mucilage secretion and the movements of blue-green-algae. Protoplasm 1968, 65, 223–238). This means that the sense of rotation may only be responsible for the direction of the spiral but not for its emergence in the first place. It would be good if the authors would distinguish between these two phenomena and discuss them accordingly.	1	2	life4040819_makarova	1
+The current paper is focused on Phormidium. We never used Myxobacteria, and we have no idea about the motility in Myxobacteria. To clarify the situation, the mention to Myxobacteria was added in the text.	2	1	(d) In Lines 309–315: References 17 and 22 are accurately described, however these models relate to gliding in Myxobacteria, a distinction not noted in the text. Thus, comparisons between these models (including the focal adhesion model) and the current model are only valid if the mechanism of motility in these microorganisms is the same. The authors should (briefly) explain their reasoning on the validity of comparing these two systems, and the implications of their findings to motility in Myxobacteria.	1	2	life4040819_makarova	1
+Lines 94–95: We have determined genomic sequences of many organisms, but it is not easy to publish the genomes as genome paper. The sequences should be connected by PCR, and annotated. In the current study, we are interested in the genes involved in motility. We annotated the related genes but we would not connect the contigs and annotate all the genes. Nowadays, every researcher can sequence his/her own materials quite easily. It is not necessary to deposit all the raw sequence data.	2	1	Lines 94–95: Is it standard to sequence an entire genome, but only deposit several clusters, or should the entire genome be accessible to allow other researchers access to this source?	1	2	life4040819_makarova	1
+Lines 227–230: This was corrected.	2	1	Lines 227–230: The interpretation of the counterclockwise spirals here is somewhat distracting. It can be left as an observation here, and the model explained later.	1	2	life4040819_makarova	1
+Lines 316–317: We did not notice this point. Polysaccharide chain may not be very long. Many of the products of the hps gene cluster encode glycosyltransferases and pseudopilins, which are, respectively, involved in the synthesis and secretion of the slime. The secretion of slime is likely mediated by some molecular machinery that is otherwise involved in type II secretion/motility machinery. Reference 24 was added.	2	1	Lines 316–317: This model for slime secretion suggests that rather than slime being comprised of a single, long, polysaccharide, it is made of smaller subunits that are secreted in a step-wise fashion. The authors should connect this idea to the finding that pseudopilins appear to be part of the molecular machinery, as the model and the data are coherent on this point.	1	2	life4040819_makarova	1
+We thank the reviewers for giving us helpful comments. This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript. The text was revised according to the comments. In addition, Figure 3A was replaced by a figure of better contrast. Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement. Within the text, corrected words are highlighted.	2	1	A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.	1	2	life4040819_perova	1
+We are not sure if type IV pili are visible in our EM figures. They are very thin (less than 1 nm) and normally visualized by negative staining. In the published EM figures of Phormidium (conventional and cryo EM), type IV pili have not been noted. The filaments that we see in our EM figures could be components of sheath materials.	2	1	As depicted in some of the electron micrographs in Figure 3, the authors describe the presence of thin filaments within the sheaths surrounding the motile filaments. Is it possible that these are in fact type IV pili? How does the structure compare to other published reports of type IV pili?	1	2	life4040819_perova	1
+This was corrected.	2	1	In Section 3.5, Analysis of Movement of Filaments, the descriptions were confusing at times. For instance Line 253 states “only in some exceptional cases (the straight line in Figure 6D), net displacement was achieved”. Net displacement presumably refers to any final position after a cycle that differs from the starting position. In other words, net displacement would refer to any value other than zero in Figure 6D. All of the points on this graph appear to indicate net displacement, albeit some less than others. Do the authors mean to state that only in a few instances was a large net displacement achieved? Otherwise the authors’ statement does not appear correct. Perhaps using numerical values instead of statements such as “the average velocity was quite limited” would help to clarify.	1	2	life4040819_perova	1
+We did not find such an indication in the literature. This point was added in the text.	2	1	The authors observe that the velocity of movement was maximal immediately after a reversal and then declined exponentially until the next reversal. The authors then suggest that this observation is theoretically inconsistent with type IV pili or focal adhesion complexes as the driving force for movement. Are there any available published reports quantifying the motility of organisms using these systems that would corroborate this empirically? In short, has it been demonstrated that bacteria moving by twitching motility or focal adhesion do not show a similar pattern for velocity of movement?	1	2	life4040819_perova	1
+The expression was modified.	2	1	The authors state that supracellular structures emerge from unordered motion and are therefore not encoded for genetically. While their model for spiral formation does not require a genetic program for controlling motility, it may not always be the case for other supercellular structures. In other words, it may not always be true that supercellular structures are not genetically determined.	1	2	life4040819_perova	1
+Line 45—This was corrected.	2	1	Line 45—suggest adding “to” before “back-and-forth motion...”	1	2	life4040819_perova	1
+ Lines 268 and 269—These were corrected.	2	1	Lines 268 and 269—“(D)” and “(E)” should be bolded to be consistent	1	2	life4040819_perova	1
+ Line 287—Two values were added.	2	1	Line 287—appears to be missing e-values. The beginning of this sentence indicates 10 genes and then only gives 8 respective e-values for the 10 genes.	1	2	life4040819_perova	1
+ Line 290—Some description was added.	2	1	Line 290—do orf303 and orf297 have homology to any characterized proteins? Are they conserved hypotheticals? Is there any indication of the function for these?	1	2	life4040819_perova	1
+ Line 415—This was corrected.	2	1	Line 415—“single” appears to be formatted differently than the rest of the text	1	2	life4040819_perova	1
+The choice of EM technique might have been wrong in visualizing the so-called oscillin fibrils, but was effective in visualizing membrane structures. The absence of “oscillin” was based on the genomic analysis but not on EM images. In fact, “oscillin” is a glycine-rich large protein which is poorly conserved in bacteria. In the reported case of P. uncinatum, oscillin might be important in forming fibrils, but in other organisms, other proteins could function as surface fibrils aligning the flow of slime. We agree that the method of fixation was not good for preserving surface structures such as oscillin fibrils, if present.	2	1	About 20 years ago it was shown that in order to preserve crucial ultrastructural features of cyanobacterial cell walls, the cells have to be processed using cryo-procedures (Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395). The authors use an outdated and very artifact-laden method to process their samples. This choice is responsible for a number of shortcomings that need to be addressed: The lack of certain features such as the S-layer and the oscillin fibrils in the reported pictures are attributable to the choice of preparation, as they are clearly present in two species of the same genus described in the above cited paper. Moreover, these structures would explain the rotation of the cells that now is somewhat “mysteriously” explained by a helical slime ejection that is not supported by the observed arrangement of the nozzles. Therefore the authors should do the following: Response:	1	2	life4040819_perova	1
+This point was added in Section 3.7, in which oscillin is discussed. In general, pore-inclination and oscillin fibrils are not exclusive.	2	1	Clearly spell out in the text why there are no oscillin fibrils and surface structures visible and compare their results with the published results of the cell wall structure of the species of the same genus prepared using cryo-procedures.	1	2	life4040819_perova	1
+There is no clear distinction in the text between the “sheath” and the secreted “slime” of the cells, which are two completely different structures both physically and chemically (Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Figure 1A shows a filament that lacks the “sheath”, while in other pictures the structure is visible. The reason for this is the following: cyanobacteria of the genus Phormidium build over time a carbohydrate layer on their surface that is physically attached to their cell surface, usually called the sheath. Filaments that are ensheathed are non-motile! In contrast, the slime that is secreted by gliding filaments of the same species is not visible in TEM preparations, even when cryo-preservation methods are used (see Figure 1A of the ms and Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395 and Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Therefore, the authors should go through their ms and make a careful distinction between these two structures. For example, in the discussion it sounds as if the “sheath” is preventing the R5 cells from sliding, however this is the slime tube that is secreted that is not physically attached to the cell surface at all.	2	1	The quality of Figure 1A was rather bad. The contrast was corrected and a new figure was inserted. In the new Figure 1A, the sheath is quite visible. The quality of EM figure is very low as embedded in an MS word file. We provide a better quality figure as supplemental Figure S1. The use of “sheath” is therefore correct in the paragraph describing the figure. In the model, the word “sliding” was not understood in the sense in which we wanted it to be. This is lateral sliding that provokes curvature of the filament. This point was clarified in the text.	1	2	life4040819_perova	1
+What is “the basal body part”? Basal body is an organelle of eukaryotic cell. If this means the connection of junctional pore tube and inner membrane, it is clearly seen in the figure, as well as in old paper by Halfen and Castenholz. A high quality figure is provided as Supplementary Figure S1. The “bulges” in the inner membrane are also visible. In the Line 183, the description was revised.	2	1	Please omit the description of the basal body part of the junctional pores or provide substantially better pictures. Even after careful inspection of your images, I cannot convince myself of seeing these parts. Moreover, in cryo-preserved and freeze-fractured cells there are no such structures visible. That does not mean that they don’t exist, it may only mean that none of the so far used preservation methods reveals these structures.	1	2	life4040819_perova	1
+In Figure 3D, the junctional pores are seen as an array of tilted tubes. The mechanism of rotation is still a mystery because oscillin itself is not present in the strain KS. This is discussed in Section 3.7 with reservations on our hypothesis of inclined junctional pores. On the other hand, the involvement of fibrils in the rotation and locomotion was proposed a long time ago, whether the fibrils are made of oscillin or other proteins. In this respect, we still have to work a lot to identify the real mechanism of rotation.	2	1	Please omit the description about helical tilted junctional pores. These structures are clearly not arranged in this way and the reasons for rotation are the unfortunately not-preserved surface structures mentioned above.	1	2	life4040819_perova	1
+This is not a good way of naming headings. A heading should not be a phrase or conclusion, but should be descriptive words.	2	1	Results section headings and figure captions should be phrased as conclusions, supported by the data presented, when appropriate (can be a minor change, for example 3.2 could be titled, “Phormidium colonies form divergent structures,” or larger, for example 3.5 could be titled, “Individual filaments undergo directional reversals, with maximum velocity immediately following a reversal.”	1	2	life4040819_perova	1
+This was described in Section 3.7.	2	1	Please clarify the source of the helical flow of the slime, which is most likely due to the presence of the not-preserved helically arranged cell surface proteins not the arrangement or tilt of the junctional pores.	1	2	life4040819_perova	1
+This point was corrected.	2	1	In the model that describes the potential activity of the junctional pores (How myxobacteria glide. Curr. Biol. 2002, 12, 369–377) it is not suggested that the nozzle actually “contracts” as mentioned in the ms. The swelling of the slime material in the nozzle fills the nozzle and eventually will generate a counter-force of the nozzle walls of the nozzle that results in the ejection of the slime. Please rephrase the text accordingly.	1	2	life4040819_perova	1
+There might be a misunderstanding in the first part. The rotation of filament does not drive macroscopic rotation or spiral formation. What we describe in the text was the switch to turn to the left was governed by the filament rotation, but the real formation of a spiral is driven by the locomotion of the filament. The cited paper described the clumping of Anabaena cylindria in a dense culture. The clumping or aggregation in Arthrospira (Ohmori group) is mediated by cAMP, but A. cylindrica might be different. I have been using Anabaena for about 40 years, but I have never seen Anabaena filaments form a spiral on agar plates. Spiral formation and clumping are different phenomena. It is difficult to discuss the relationship (if any) between clumping and rotation.	2	1	When the cells form spirals, there are two components to this process: the formation of spirals and their macroscopic sense of rotation: clock- or counterclockwise. In the discussion it is assumed that the rotation of the filaments is crucial for both of these components. If this would be true, then no spirals should be observed in non-rotating filamentous gliding cyanobacteria. However, this is not the case. Anabaena spec. a non-rotating species can generate spirals (see Figure X; Mucilage secretion and the movements of blue-green-algae. Protoplasm 1968, 65, 223–238). This means that the sense of rotation may only be responsible for the direction of the spiral but not for its emergence in the first place. It would be good if the authors would distinguish between these two phenomena and discuss them accordingly.	1	2	life4040819_perova	1
+The current paper is focused on Phormidium. We never used Myxobacteria, and we have no idea about the motility in Myxobacteria. To clarify the situation, the mention to Myxobacteria was added in the text.	2	1	In Lines 309–315: References 17 and 22 are accurately described, however these models relate to gliding in Myxobacteria, a distinction not noted in the text. Thus, comparisons between these models (including the focal adhesion model) and the current model are only valid if the mechanism of motility in these microorganisms is the same. The authors should (briefly) explain their reasoning on the validity of comparing these two systems, and the implications of their findings to motility in Myxobacteria.	1	2	life4040819_perova	1
+Lines 94–95: We have determined genomic sequences of many organisms, but it is not easy to publish the genomes as genome paper. The sequences should be connected by PCR, and annotated. In the current study, we are interested in the genes involved in motility. We annotated the related genes but we would not connect the contigs and annotate all the genes. Nowadays, every researcher can sequence his/her own materials quite easily. It is not necessary to deposit all the raw sequence data.	2	1	Lines 94–95: Is it standard to sequence an entire genome, but only deposit several clusters, or should the entire genome be accessible to allow other researchers access to this source?	1	2	life4040819_perova	1
+Lines 227–230: This was corrected.	2	1	Lines 227–230: The interpretation of the counterclockwise spirals here is somewhat distracting. It can be left as an observation here, and the model explained later.	1	2	life4040819_perova	1
+Lines 316–317: We did not notice this point. Polysaccharide chain may not be very long. Many of the products of the hps gene cluster encode glycosyltransferases and pseudopilins, which are, respectively, involved in the synthesis and secretion of the slime. The secretion of slime is likely mediated by some molecular machinery that is otherwise involved in type II secretion/motility machinery. Reference 24 was added.	2	1	Lines 316–317: This model for slime secretion suggests that rather than slime being comprised of a single, long, polysaccharide, it is made of smaller subunits that are secreted in a step-wise fashion. The authors should connect this idea to the finding that pseudopilins appear to be part of the molecular machinery, as the model and the data are coherent on this point.	1	2	life4040819_perova	1
+The three references have been noticed with gratitude and included in the MS.	2	1	Section 1. The place of RNA in LUCA (page 2): In search of features that are more conserved (carrying deep phylogenetic memory) than the sequence of genes, Wächtershäuser focuses on a paper of his in Systematic and Applied Microbiology (1998) that uses gene content and order of microbial genomes to make inferences about the last universal common ancestor (LUCA) of cellular life. He then mines the significance of some of the conserved chromosomal segments in light of some other evidence. The exercise is at places compelling, but forgets some recent, very global and exhaustive analyses that also use highly conserved biological features to reconstruct the makeup of LUCA (e.g., gene order, 3D molecular structure, molecular functions). See for example PubMed references PMID: 17370266, PMID: 21612591, and PMID: 17908824, which are in line with some conclusions derived from the alignment of Figure 1. The fact that these other analyses make use of hundreds of genomes to infer the ancient biochemistry of LUCA complements and strengthens the preliminary and fragmentary analysis of only 19 of them by the author, which also excludes eukaryotic genomes from the set (understandably, few sequenced genomes were available in 1998, and eukaryotes are in general “master rearrangers”).	1	2	life4041050_makarova	1
+The 1998 paper leaves much to be desired. Its deficiencies reflect the excitement of the first hour. The state of the art at that time may be gleaned from an authoritative paper that came to the opposite conclusion: A.R. Mushegian and E.V. Koonin “Gene order is not conserved in bacterial evolution”. TIG 1996, 12, 289–290. The gene cluster table of 1998 was mainly retrieved from the annotations in published genomes and constructed manually with paper and pencil. The state of information technology at that time is reflcted by the fact that the table was folded an individually pasted by hand into each issue by the publisher.	2	1	Given my interest in the bioinformatics of gene content and order, I took the liberty of studying the brief Systematic & Applied Microbiology paper to check the validity of the “reconstruction” methods of Figure 1. The algorithmic implementation that is described is quite raw and does not extract important information that is embedded in the clusters of conserved gene segments. Important algorithms have been devised since the initial work of Sankoff in the sixties and seventies to do exactly that. I refer to the work of Pevzner, Tesler and Bourque as good examples, but also of Warnow. I also recommend visiting GRIMM (http://grimm.ucsd.edu/GRIMM/) and perhaps using the server to confirm or extract additional information from the alignments that are summarized in Figure 1. The Wächtershäuser algorithm makes use of conserved elements of gene content and order but discards information provided by the actual rearrangement operations that erase gene order history. The algorithm does not describe how gene homology was detected, how the limited set of genes was selected, and how the alignments were constructed (I imagine by hand). It is not clear if a guiding tree was used in the alignment (though this is mentioned in Line 65), since this is not made explicit in the 1998 publication. An alignment implies a tree but usually alignment algorithms are greedy and problematic and represent the most important limiting step of a phylogeny (the field is thus moving to the joint alignment and tree reconstruction). Therefore, I do not think the ancestors were properly reconstructed (the tracing of features in ancestor nodes of trees are not described, nor the actual trees). Regardless of all of these limitations, the tight conservation of certain segments is enough to show the existence of a core of ribosomal proteins that is universally present in cellular organisms. This in itself is valuable. Of course, a sample of 19 microbes may not be enough to encompass molecular diversity and the absence of Eukarya may also be problematic for any global evolutionary statement. In other words, the risks of sampling bias are clearly present and should be mentioned in the manuscript.	1	2	life4041050_makarova	1
+The 1998 paper contemplated speculatively a combination of small-scale gene doubling (as evidenced by the immediate neighborhood of EF-Tu/EF-G) and of a large-scale gene cluster doubling (as evidenced by the spacing between secE/secY and rpoH-A/rpoD) with the hope of a future deeper understanding based on folding structures. Now the referee makes an exciting suggestion that may, if executed successfully, go some way to satisfy that hope.	2	1	The list of common genes in the alignment of Table 1 is enriched in small and large subunit ribosomal proteins that are the most ancient of the ribosomal set, according to the Caetano-Anolles theorem that is mentioned later in the manuscript. Interestingly, the most ancient of them are clustered toward the 5’ end of the genomic sequences that were aligned (S12, S17, S5, S4 and L2, L3 and L24). Could this imply a possible ancient segmental duplication? Also interesting is the placement of the most ancient ribosomal protein S12 between exactly two polymerase (beta and beta prime) and elongation factor (entry and translocation) genes (separated by single and much more derived ribosomal proteins). Could this be an ancient memory of the ribosome mediating translation and replication? A commentary would enhance the value of the section.	1	2	life4041050_makarova	1
+The self-cleavage of RNA by 2'-OH is a chemical textbook fact. The 2'-OH group has the proper position and orientation for a nucleophilic attack on the phosphate bridge. The kinetics of the reaction is greatly favored by the 5-membered ring structure of the resulting cyclic phospho-bisester. The length of the RNA molecule is not relevant since each nucleophilic attack causes destruction of the chain. The effect should not be confused with the length-dependent “error catastrophe” of accumulating mutations of RNA. Incidentally, some anaerobic ribonucleotide reductases are ancient, while others (aerobic ones) are later inventions.	2	1	In terms of the genome organization of LUCA (Line 105), there are numerous arguments in favor of an RNA ancestral genome and the late unfolding of DNA as genetic repository (perhaps through viruses). I cannot understand how 2'-hydroxy groups could destabilize the molecule and lead to “intramolecular self-destruction” (Line 107). RNA has the wonderful property of folding in search of energetic and kinetic minima. These processes make use of a frustrated landscape, which is powerful in terms of its biological potential (stability, function, information). It is much more versatile than the rigid DNA alternative, which is enriched in the ‘information’ capacity. None of them have “self destruction” properties, which would have been weeded out by selection and self-organization very early in “chemical” evolution. In turn, everything about LUCA should be considered quite modern and far away from an initial FeS world. I refer to the work of Daniel Lundin and colleagues in Sweden about the rather late rise of some structures of the ribonuceotide reductase enzymes needed to built the DNA polymers. The Maurel theorem stands if one thinks of single genome molecules. But what if there were many short RNA genomes, as anticipated by Woese, each perhaps linked to different and ancient tRNA-like cofactors? As the author mentions, LUCA is contemporary to a rather complex biochemistry, with numerous structures harboring a multitude of active and allosteric sites in proteins, most of which drive central enzymatic functions.	1	2	life4041050_makarova	1
+Genome rearrangements are certainly important during later evolution of the phyla. At the level of LUCA, i.e., prior to the splitting of the domains, it is not clear, if and to what extent rearrangements of the modern style occurred. In this regard we should bear in mind that the LUCA genome may have exhibited sense-antisense coding on both strands as suggested by Rodin and Carter. This position has been adopted and discussed in the present paper. Therefore, speculations concerning possible genome rearrangements may be a bridge too far.	2	1	In Line 114, the role of chromosomal rearrangement in evolution is not well described. What are the effects of genome rearrangements? Several scenarios are possible, including: (1) Genes remain linked R4 probably because it is more efficient to transcribe genes that produce interacting proteins (part of complexes) than those that are not; (2) Genes remain linked if they are of relatively recent origin (rearrangement has not had the change to split them apart); (3) Genes remain linked if they are part of functional groups historically united by genomic regions (encoding metabolic functions or rRNA? ); this includes genes sharing an operon structure for economy purposes in highly reduced organisms such as bacteria; (4) Genes remain linked because they originated when genomic rearrangements were not biochemically motivated and their sequence makeup was later refractory to rearrangement hotspots. There is a rich literature about rearrangement, hotspots and many processes related to these (including domain organization in proteins). Some discussion in this front could be clarifying.	1	2	life4041050_makarova	1
+The present analysis comes to the conclusion that the sets of canonical amino acids and bases as well as the genetic triplet code were largely complete at the level of LUCA. The Wong coevolution theory of the genetic code has been discussed in detail. The literature comprises numerous proposals concerning the origin of translation and other aspects of the genetic machinery. A review of all these proposals and many others is beyond the scope of the present paper. The present paper is a research paper and not a review paper. It aims at a comprehensive account of early evolution from the origin of life all the way to LUCA. This puts a systematic constraint on literature selection. An effort (unfortunately fallible) has been made to include all those references that integrate with the main lines of the present account into a coherent account. Contributions by others that have the character of theoretical modules that fit well into this account have been termed “theorems” with names of the main authors attached. A reference to a paper on nucleotide biosynthesis phylogeny is now cited in Section 7.	2	1	In relation to comments of Line 188 onwards, Di Giulio, Caetano-Anolles and others have suggested that the genetic code started to unfold prior to LUCA but continue to do so once life diversified. The corollary, is that the complete canonical set of amino acids may have not been encoded in LUCA. In terms of metabolic and biochemical competency, there are numerous and interesting studies, which have not been cited (especially in origins and evolution of modern metabolism). This includes the coevolution theory of the genetic code, the coexistence of prebiotic chemistries with modern metabolic reactions (recently studied for nucleotide metabolism), and theories about the origin of translation.	1	2	life4041050_makarova	1
+Based on the valuable criticism Section 2 has been extensively revised. Terminology has been clarified. This Section has a rather restricted purpose. It provides chemical arguments for the proposition that the pioneer organisms could only exist at high temperature and that the subsequent forms of life remained hyperthermophilic for a long time until much later an irreversible evolution generated organisms that required lower and lower temperatures. This conclusion places severe constraints on all aspects of the evolution of the genetic machinery. The fascinating topics of thermodynamics, energy dissipation and information are outside the scope of the paper.	2	1	Section 2: Thermal course of evolution (Page 6): Why a focus on thermal energy? The framework should be on thermodynamics, energy dissipation and information, all of which are linked. What is thermally upward or downward adaptation? Is it conquering niches on Earth or a process involving molecular makeup? Vocabulary and definitions are murky, especially related to the links of environmental thermal fluctuations, energy of folding and stability of polymers. Subsection (1) must be rewritten to help the reader understand the ideas. Should all adaptations comply with maximizing energy R5 dissipation? How can this be reconciled with upward and downward trends? No references are provided despite the rich literature underscoring the controversial link between physics, information theory and biology. If the focus is conquering planetary niches, then perhaps reference the contrasting views and some of their proponents (deep sea versus surface; thermophilic versus mesophilic, etc.). Further elaboration of the Wolfenden theorem appears relevant and its connections to what is known about the origin and evolution of metabolism also of importance.	1	2	life4041050_makarova	1
+The author expresses his gratitude for the two additional references, which have been included in the text.	2	1	Section 5. Place of RNA in the origin and early evolution of translation/Pre-translational coding of peptides. The view that is presented in this crucial segment posits a pre-translational mechanism (side-by-side tRNA mediated coding) that very much resembles the ribosomal entropic system, which could be very advanced. What if the pre-translational coding was assembly line-like and mediated by the aminoacyl-tRNA synthetases themselves? See PMID: 23991065 for one such alternative and a related previous model (PMID: 22210458). This would also match commentary of a possible early origin of non-ribosomal peptide synthesis machinery compared to that of the ribosome.	1	2	life4041050_makarova	1
+The statement has been clarified.	2	1	Section 9. General overview. While the grand finale “Nothing in evolution makes sense except in the light of chemical predetermination” is impressive and summarizes the championing work of the author, chemical predetermination may apply to the very early stages of evolution and not to many of the stages described in the manuscript. How much chemical predetermination can there be with polymers as these explore a minute fraction of the space they make possible? In the enthalpic-entropic gradient that is proposed, there is also a gradient from “predetermination” to historical contingency. The boundaries of such a gradient is murky and the final statement may not apply to much of modern biochemistry, once proteins start to achieve stable complex structure and much earlier that the time of LUCA. In fact, the genome rearrangements that underlie Figure 1 are testament to the historical contingencies that were already at play in LUCA and not to chemical predetermination.	1	2	life4041050_makarova	1
+The author prefers the present title, because the paper is concerned with the origin of the genetic machinery. Theories on the origin of life or on other aspects of early evolution, such as cellularization, serve merely as starting points.  In the introduction the term “or proteins” has been added after “RNA” ,  and the term “retrodict” has been defined.  The term “interpolate” has been clarified.  The problem of mutational saturation is now included in the discussion of Figure 1.  The term “multiply impaired” has been replaced by a clearer wording.  LUCA is discussed only in Section 1.  The protein cycle is discussed in a separate Section 4, which is concerned with the course of evolution before LUCA.	2	1	Line 406. “LUCA protein cycle”. Why bring LUCA to a link with the proposed “protein cycle”. LUCA is just the last of a chain of ancestor of diversified life. If the protein cycle requires a coupling of the enthalpic activation of amino acids and the entropic biosynthetic action of a primordial ribosome, then perhaps it is better to call it “ribocellular protein cycle”, since this coupling marks the start of modern cells, not necessarily “modern diversified cells” (i.e., LUCA).	1	2	life4041050_makarova	1
+There are many relevant literature references. Among these there are specific proposals concerning a circumscribed problem that are included to fill a logical gap, as a theoretical module so to speak. These have been designated by the term “theorem” with the added name(s) of the main R7 author(s). This makes it clear that the account given is comprehensive in the sense that major independent contributions by other scientists integrate readily with the overall account given.	2	1	Please explain what do you mean with Kandler, Wolfenden etc. Theorem. Is a theorem Kandler’s theory or proposal?	1	2	life4041050_makarova	1
+The relationship of His and Trp biosyntheses has been toned down.	2	1	Line 716: The biosyntheses of His and Trp are not so closely related. They only have in common that they use phosphoribosediphosphate as substrate and therefore share two related enzymes.	1	2	life4041050_makarova	1
+Section 8 has been revised in order to address the issues involved in the last query.	2	1	In the iron sulfur world the surface metabolism prevents the diffusion of the fixed products into the ocean. But without compartmentation soluble intermediates would escape. Furthermore, the establishment of early bioenergetics based on electrochemical Na+ gradients would have been impossible.	1	2	life4041050_makarova	1
+The concern the reviewer mentions here was also one of our main conclusions for the article. Apparently we did not formulate it well enough. It has thus been reformulated to: “However an accurate taxonomy can never be achieved by the use of a barcoding method only, since it is based on nucleotide substitutions of a single gene. Accurate classification always requires an integrative taxonomy effort R2 including characteristics from ecology, morphology and physiology, as already previously suggested for prokaryotes [9,28,29,45], and as it is becoming common also for animal taxonomy (e.g., [53,54]).” Moreover, it is necessary to mention the following aspects of this manuscript: (1)	2	1	The manuscript of Eckert et al., submitted into the journal Life, concerns in principle again only one, but very important genetic criterion for evaluation of prokaryotic diversity. It should be a serious part of this complex evaluation. It is very important and useful and publication of the article is surely recommendable. However, the connection, coincidences and relations to other criteria should be better expressed.	1	2	life5010050_makarova	1
+We understand that our choice of naming as in the database needed clarification. We have thus added the following paragraph. We thank the reviewer for the useful example and have included it: “The names of genera and species used in this study where copied form the names given in the database. This choice was made for the reason that cyanobacterial phylogeny underlies continuous changes and genera and species are often reclassified (e.g., [34]). Thus a whole different type of work would be required in order to use all state-of-art classification of cyanobacteria. Moreover, for the aims of this study not so much the phylogenetic classification of the bacterial species, but the genetic structure of the cyanobacterial 16S rRNA genes was of importance. The use of the names provided in the database on the other hand enables other researchers to conduct a similar analysis using the same sequences. Thus some old and revised names are used, for example the species name Anabaena bergii is used throughout the study despite its revised taxonomy in the genus Crysosporum [35].” (2)	2	1	If we accept also strains and populations from literature or from databases to the evaluation, there is always questionable the taxonomic determination (scientific names) of various species and strains, which are based on the old nomenclature and morphology. The critical revision of identification of these strains and cited materials is necessary.	1	2	life5010050_makarova	1
+We understand that particularly the case of Synechoccocus needs clarification and have thus extended the explanation there: “Sequences termed Synechococcus on the other hand were analysed together with sequences of the monophyletic clade Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included, too”.	2	1	It follows from the whole article that the authors accepted the names (and also concepts) of taxa (mostly genera) from older literature, without necessary criticism. There are included revised genera according to modern methods together with taxonomically very problematic groups. As examples is possible to mention that Aphanizomenon ovalisporum and Anabaena bergii are really very related taxa, but they were already re-classified in one special and separate genus Chrysosporum. On the other hand, the complex “Prochlorococcus/Synechococcus” is still very unclear and surely is distant from the typical genus Synechoccocus. etc.	1	2	life5010050_makarova	1
+We agree with the reviewer that we have to avoid misunderstandings of this kind and added a very clear statement to the discussion: “Moreover it has to be emphasised that a close analysis of the actual properties of the cyanobacterial groups tested is limited by the fact that we used the provided R3 names for the sequences and groups. Thus this study intends only to verify the existence of a barcoding gaps within the 16S rRNA sequences of certain cyanobacterial groups, and by no means revise or confirm the complex cyanobacterial taxonomy.”	2	1	Numerous cited taxa (on the generic level) were already controlled and revised by most modern methods, and examples in the manuscript should be used especially and selected from these genera. It is true, that majority of the cited genera were already revised (Microcystis, Planktothrix, Aphanizomenon), but just these taxa are important as examples and should be preferred. From the presented results follows only the confirmation of the present methodological principles and a modern system. This fact should be emphasized.	1	2	life5010050_makarova	1
+We agree with the reviewer that we have not stressed this fact enough and have added more sentences explaining why OTUs are used and the problem: “Enormous progress happened in prokaryote taxonomy in the recent years [8,12,24]. However, the data produced with novel methodologies, such as next generation sequencing, often requires a high throughput taxonomic classification of sequences such as fixed threshold to identify OTUs. This kind of fixed numeric classifications can always only be a vague approximation to the actual structure of relatedness of organisms.”	2	1	It is necessary to stress the fact that the numerical criteria (e.g., exactly limited percentage similarity) never are valid uniformly for large groups of organisms and cannot be applied as one main criterion in large groups.	1	2	life5010050_makarova	1
+our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gap existed at all.	2	1	As concerns the delimitation of taxa on the level of cyanobacterial species, there exists a lot of valuable literature, which was not cited. The characterization and separation of species was not satisfactory solved, but it is particularly important (criteria are not unique for all genera) and need better discussion.	1	2	life5010050_makarova	1
+we now include new citations in relationship to different problems: Butlin et al. (2009), Cohan (2011), Cohan (2013), Cohan & Aracena (2012), Diekmann et al. (2004), Nosil (2012), Vos (2011), Wiedenbeck & Cohan (2011).	2	1	In Line 40 is cited only DeQueiroz (2007) to the problematic of species concept. It would be useful to evaluate and mention many other authors, who discussed this question more complexly. It concerns also several other problems, e.g., problematics of horizontal gene transfers (Line 43), “bounderies between taxon units” (Line 45), species concept (Line 48), etc. About species concept in cyanobacteria exist particularly many studies from last years.	1	2	life5010050_makarova	1
+We agree with the reviewer’s concern. In fact, this is exactly the reason why we chose Cyanobacteria for this analysis. We reformulated this part of the introduction, to make this statement more clear, which now reads: “We choose Cyanobacteria as an example of prokaryotes, not because they are representative for all prokaryotes, but because there is ample phenotypic, ecological, physiological ultrastructural, and biochemical evidence of the existence of independently evolving units in this group [20,21]. Thus, the expectation is that, if a barcoding gap exists in prokaryotes, this should be more easily seen in taxa where groups can be identified also with other methods, as in Cyanobacteria.”	2	1	I am not sure that cyanobacteria can be accepted as a typical example of all prokaryotes. On the contrary, they have very special position (phylogenetic, metabolic, function in nature, ultrastructural, they grow often in multicellular and differentiated thallus) and their diversity must be evaluated respecting these specificities. It is clear that the taxonomic classification must be different in different groups of organisms.	1	2	life5010050_makarova	1
+We apologize for our carelessness in double-checking the naming of the organisms and hope that we now eliminated all errors.	2	1	The correct citation of scientific names is the obvious request of serious scientific publications. Few unnecessary mistakes are in the manuscript, e.g., in the whole text and also in Figures is written “Arthospira” instead the correct Arthrospira, on other places must be Leptolyngbya (Table 1, Line 191), Fischerella (Line 207), Aphanizomenon (Line 218), Cylindrospermopsis (Line 219).	1	2	life5010050_makarova	1
+We agree with the reviewer, this part was formulated clumsily. What we meant was actually exactly what the reviewer means. The two groups were polyphyletic and we could only chose a monophyletic group when analysing them together. We changed the sentence to: “… by two genera that were monophyletic only when taken together in the database used, e.g., Leptolyngbia and Chamaesiphon …” In accordance we did not check for relatedness of the taxa, neither in this one nor for any other one.	2	1	I do not understand why Leptolyngbya and Chamaesiphon could be “closely related genera”. It is in contraversion to the whole up to date results and taxonomic classification. The authors really found the “close” relation just between these two genera? By way, they both are polyphyletic and will be surely divided in several different taxa after the following precise studies.	1	2	life5010050_makarova	1
+We agree that this might be confusing to the reader. We have therefore clarified this choice and added this part to the first M&M section and the literature suggested has been cited: “Sequences termed Synechococcus on the other hand were analysed together with sequences of Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included too.”	2	1	It is overall well-written and innovative work, but there are few things to consider in methods and results, which should be revised, and some formal flaws. When you refer to the Synechococcus, it is important to specify, which clade you have in mind. Honda et al. (1999) and Roberston et al. (2001) showed Synechococcus is polyphyletic genus composed of a least 5 lineages. There are more recent works showing even more clades. From an amount of sequences and relationship to Prochlorococcus, I can assume, it is a marine pelagial picoplanktic clade. However, it should be specified for unexperienced reader in the field of cyanobacteria. The other polyphyletic genera should be also specified—Phormidium, Leptolyngbya, Microcoleus and others. Moreover, although the sequences might have same name (i.e., Synechococcus), they belong to polyphyletic groups, thus they probably belong to different genera, which have not yet been described.	1	2	life5010050_makarova	1
+The sequences we used were clustering monophyletically a-priori in the tree provided in the database used, and we chose them independently of the name. We now clarified this in the first section of M&M: “Thereby a group was considered monophyletic if it was monophyletic in the tree provided with the database, regardless of the taxonomy and the nomenclature of the organisms included in the clade (Supplemetary Figure 1), and only secondarily if there was a correspondence with known taxa.”	2	1	They might also appear to be monophyletic, because there are missing taxa between them.	1	2	life5010050_makarova	1
+As mentioned above, we did not only choose sequences that had the same name, but groups of sequences that were monophyletic in the database tree. Thus we should not have artificially introduced a barcoding gap by sequence selection.	2	1	This might also cause false positive barcoding gaps, because there would be low similarity among sequences with same name. Therefore it may largely affect results.	1	2	life5010050_makarova	1
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names. We thus only briefly discuss names and taxonomy in our analysis. Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point. We hope that now both reviewers, and all other potential readers, will not be misled by our approach. We do not deal with taxonomy at all, only with testing the presence of a barcoding gap. Similarly to the replies for the previous reviewer, our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gaps existed at all. Given this rationale and the kind of data we used, no clear statement towards the two barcoding groups within Cylindrospermopsis can be made. Nearly all sequences within the database are termed Cylindrospermopsis raciborskii by the people who deposited the sequences. However the clade does not contain the type-strain (which was not in the database). So we would rather not speculate too much on the taxonomic naming of these and other barcoded groups. However, we can clearly say that what is deposited in database SILVA 111 as Cylindrospermopsis contains two species with a barcoding gap. For Planktothrix we added a sentence: “Considering the naming of Planktothrix sequences in the database, some OTUs and ABGD units seem to correspond not only to monophyletic groups but also to named species such as P. mougeotii or similarly in the case of Fischerella muscicola (Figure 3).” R6 However we wish to remain careful on these kinds of statements since we are not sure if the underlying naming of the species used is correct or not.	2	1	I suggest that the paper should be expanded with a discussion, whether barcode gaps are able to delimit described species. For example, are there two species within Cylindrospermopsis? There seems to be a barcoding gap within this clade. In this particular case, it would be beneficial to use a species epithet too to avoid confusion. Is it Cylindrospermopsis raciborskii? Does a barcoding gap in the Planktothrix clade correspond to the described species?	1	2	life5010050_makarova	1
+We understand why the reviewer would think that that would be beneficial. However, one has to keep in mind that we were not constructing a phylogenetic tree for all cyanobacteria but for single genera. In this case it is preferential to use a sister group that is more closely related to gain resolution within that group. e.g., Hedtke, S.M. ; Townsend, T.M. ; Hillis, D.M. Resolution of phylogenetic conflict in large data sets by increased taxon sampling. Syst. Biol. 2006, 55, 522–529. Moreover, for the sake of the barcoding analyses, the choice of the outgroup will not affect the results, given that it will not change the relationships towards the tips of the tree.	2	1	Why did you use Phormidium as an outgroup (Line 139)? Would not be more beneficial to use Gloeobacter or some other bacterium?	1	2	life5010050_makarova	1
+The last paragraph of the discussion already included some comments on this issue. Now the paragraph has been expanded to provide a more detailed discussion on the possibility of using a DNA barcoding gap in cyanobacteria.	2	1	Is there any way to use barcoding gaps in a taxonomy of cyanobacteria? This might be added to discussion.	1	2	life5010050_makarova	1
+We apologize for our carelessness in double checking the naming of the organisms and hope that we now eliminated all errors.	2	1	Minor point: There are errors in nomenclature throughout the text, figures, and supplements, e.g. Leptolyngbya, Aphanizomenon, Arthrospira. You might check: http://www.cyanodb.cz/. It is an updated database of names of cyanobacteria.	1	2	life5010050_makarova	1
+The concern the reviewer mentions here was also one of our main conclusions for the article. Apparently we did not formulate it well enough. It has thus been reformulated to: “However an accurate taxonomy can never be achieved by the use of a barcoding method only, since it is based on nucleotide substitutions of a single gene. Accurate classification always requires an integrative taxonomy effort R2 including characteristics from ecology, morphology and physiology, as already previously suggested for prokaryotes [9,28,29,45], and as it is becoming common also for animal taxonomy (e.g., [53,54]).”	2	1	The manuscript of Eckert et al., submitted into the journal Life, concerns in principle again only one, but very important genetic criterion for evaluation of prokaryotic diversity. It should be a serious part of this complex evaluation. It is very important and useful and publication of the article is surely recommendable. However, the connection, coincidences and relations to other criteria should be better expressed.	1	2	life5010050_perova	1
+We understand that our choice of naming as in the database needed clarification. We have thus added the following paragraph. We thank the reviewer for the useful example and have included it: “The names of genera and species used in this study where copied form the names given in the database. This choice was made for the reason that cyanobacterial phylogeny underlies continuous changes and genera and species are often reclassified (e.g., [34]). Thus a whole different type of work would be required in order to use all state-of-art classification of cyanobacteria. Moreover, for the aims of this study not so much the phylogenetic classification of the bacterial species, but the genetic structure of the cyanobacterial 16S rRNA genes was of importance. The use of the names provided in the database on the other hand enables other researchers to conduct a similar analysis using the same sequences. Thus some old and revised names are used, for example the species name Anabaena bergii is used throughout the study despite its revised taxonomy in the genus Crysosporum [35].” (2)	2	1	If we accept also strains and populations from literature or from databases to the evaluation, there is always questionable the taxonomic determination (scientific names) of various species and strains, which are based on the old nomenclature and morphology. The critical revision of identification of these strains and cited materials is necessary.	1	2	life5010050_perova	1
+We understand that particularly the case of Synechoccocus needs clarification and have thus extended the explanation there: “Sequences termed Synechococcus on the other hand were analysed together with sequences of the monophyletic clade Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included, too”.	2	1	It follows from the whole article that the authors accepted the names (and also concepts) of taxa (mostly genera) from older literature, without necessary criticism. There are included revised genera according to modern methods together with taxonomically very problematic groups. As examples is possible to mention that Aphanizomenon ovalisporum and Anabaena bergii are really very related taxa, but they were already re-classified in one special and separate genus Chrysosporum. On the other hand, the complex “Prochlorococcus/Synechococcus” is still very unclear and surely is distant from the typical genus Synechoccocus. etc.	1	2	life5010050_perova	1
+We agree with the reviewer that we have to avoid misunderstandings of this kind and added a very clear statement to the discussion: “Moreover it has to be emphasised that a close analysis of the actual properties of the cyanobacterial groups tested is limited by the fact that we used the provided R3 names for the sequences and groups.	2	1	Numerous cited taxa (on the generic level) were already controlled and revised by most modern methods, and examples in the manuscript should be used especially and selected from these genera. It is true, that majority of the cited genera were already revised (Microcystis, Planktothrix, Aphanizomenon), but just these taxa are important as examples and should be preferred. From the presented results follows only the confirmation of the present methodological principles and a modern system. This fact should be emphasized.	1	2	life5010050_perova	1
+We agree with the reviewer that we have not stressed this fact enough and have added more sentences explaining why OTUs are used and the problem: “Enormous progress happened in prokaryote taxonomy in the recent years [8,12,24]. However, the data produced with novel methodologies, such as next generation sequencing, often requires a high throughput taxonomic classification of sequences such as fixed threshold to identify OTUs. This kind of fixed numeric classifications can always only be a vague approximation to the actual structure of relatedness of organisms.”	2	1	It is necessary to stress the fact that the numerical criteria (e.g., exactly limited percentage similarity) never are valid uniformly for large groups of organisms and cannot be applied as one main criterion in large groups.	1	2	life5010050_perova	1
+our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gap existed at all.	2	1	As concerns the delimitation of taxa on the level of cyanobacterial species, there exists a lot of valuable literature, which was not cited. The characterization and separation of species was not satisfactory solved, but it is particularly important (criteria are not unique for all genera) and need better discussion.	1	2	life5010050_perova	1
+we now include new citations in relationship to different problems: Butlin et al. (2009), Cohan (2011), Cohan (2013), Cohan & Aracena (2012), Diekmann et al. (2004), Nosil (2012), Vos (2011), Wiedenbeck & Cohan (2011).	2	1	In Line 40 is cited only DeQueiroz (2007) to the problematic of species concept. It would be useful to evaluate and mention many other authors, who discussed this question more complexly. It concerns also several other problems, e.g., problematics of horizontal gene transfers (Line 43), “bounderies between taxon units” (Line 45), species concept (Line 48), etc. About species concept in cyanobacteria exist particularly many studies from last years.	1	2	life5010050_perova	1
+We agree with the reviewer’s concern. In fact, this is exactly the reason why we chose Cyanobacteria for this analysis. We reformulated this part of the introduction, to make this statement more clear, which now reads: “We choose Cyanobacteria as an example of prokaryotes, not because they are representative for all prokaryotes, but because there is ample phenotypic, ecological, physiological ultrastructural, and biochemical evidence of the existence of independently evolving units in this group [20,21]. Thus, the expectation is that, if a barcoding gap exists in prokaryotes, this should be more easily seen in taxa where groups can be identified also with other methods, as in Cyanobacteria.”	2	1	I am not sure that cyanobacteria can be accepted as a typical example of all prokaryotes. On the contrary, they have very special position (phylogenetic, metabolic, function in nature, ultrastructural, they grow often in multicellular and differentiated thallus) and their diversity must be evaluated respecting these specificities. It is clear that the taxonomic classification must be different in different groups of organisms.	1	2	life5010050_perova	1
+We apologize for our carelessness in double-checking the naming of the organisms and hope that we now eliminated all errors.	2	1	The correct citation of scientific names is the obvious request of serious scientific publications. Few unnecessary mistakes are in the manuscript, e.g., in the whole text and also in Figures is written “Arthospira” instead the correct Arthrospira, on other places must be Leptolyngbya (Table 1, Line 191), Fischerella (Line 207), Aphanizomenon (Line 218), Cylindrospermopsis (Line 219).	1	2	life5010050_perova	1
+We agree with the reviewer, this part was formulated clumsily. What we meant was actually exactly what the reviewer means. The two groups were polyphyletic and we could only chose a monophyletic group when analysing them together. We changed the sentence to: “… by two genera that were monophyletic only when taken together in the database used, e.g., Leptolyngbia and Chamaesiphon …” In accordance we did not check for relatedness of the taxa, neither in this one nor for any other one.	2	1	I do not understand why Leptolyngbya and Chamaesiphon could be “closely related genera”. It is in contraversion to the whole up to date results and taxonomic classification. The authors really found the “close” relation just between these two genera? By way, they both are polyphyletic and will be surely divided in several different taxa after the following precise studies.	1	2	life5010050_perova	1
+We agree that this might be confusing to the reader. We have therefore clarified this choice and added this part to the first M&M section and the literature suggested has been cited: “Sequences termed Synechococcus on the other hand were analysed together with sequences of Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included too.”	2	1	When you refer to the Synechococcus, it is important to specify, which clade you have in mind. Honda et al. (1999) and Roberston et al. (2001) showed Synechococcus is polyphyletic genus composed of a least 5 lineages. There are more recent works showing even more clades. From an amount of sequences and relationship to Prochlorococcus, I can assume, it is a marine pelagial picoplanktic clade. However, it should be specified for unexperienced reader in the field of cyanobacteria. The other polyphyletic genera should be also specified—Phormidium, Leptolyngbya, Microcoleus and others. Moreover, although the sequences might have same name (i.e., Synechococcus), they belong to polyphyletic groups, thus they probably belong to different genera, which have not yet been described.	1	2	life5010050_perova	1
+The sequences we used were clustering monophyletically a-priori in the tree provided in the database used, and we chose them independently of the name. We now clarified this in the first section of M&M: “Thereby a group was considered monophyletic if it was monophyletic in the tree provided with the database, regardless of the taxonomy and the nomenclature of the organisms included in the clade (Supplemetary Figure 1), and only secondarily if there was a correspondence with known taxa.”	2	1	They might also appear to be monophyletic, because there are missing taxa between them.	1	2	life5010050_perova	1
+As mentioned above, we did not only choose sequences that had the same name, but groups of sequences that were monophyletic in the database tree. Thus we should not have artificially introduced a barcoding gap by sequence selection.	2	1	This might also cause false positive barcoding gaps, because there would be low similarity among sequences with same name. Therefore it may largely affect results.	1	2	life5010050_perova	1
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names. We thus only briefly discuss names and taxonomy in our analysis. Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point. We hope that now both reviewers, and all other potential readers, will not be misled by our approach. We do not deal with taxonomy at all, only with testing the presence of a barcoding gap. Similarly to the replies for the previous reviewer, our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gaps existed at all. Given this rationale and the kind of data we used, no clear statement towards the two barcoding groups within Cylindrospermopsis can be made. Nearly all sequences within the database are termed Cylindrospermopsis raciborskii by the people who deposited the sequences. However the clade does not contain the type-strain (which was not in the database). So we would rather not speculate too much on the taxonomic naming of these and other barcoded groups. However, we can clearly say that what is deposited in database SILVA 111 as Cylindrospermopsis contains two species with a barcoding gap. For Planktothrix we added a sentence: “Considering the naming of Planktothrix sequences in the database, some OTUs and ABGD units seem to correspond not only to monophyletic groups but also to named species such as P. mougeotii or similarly in the case of Fischerella muscicola (Figure 3).” R6 However we wish to remain careful on these kinds of statements since we are not sure if the underlying naming of the species used is correct or not.	2	1	I suggest that the paper should be expanded with a discussion, whether barcode gaps are able to delimit described species. For example, are there two species within Cylindrospermopsis? There seems to be a barcoding gap within this clade. In this particular case, it would be beneficial to use a species epithet too to avoid confusion. Is it Cylindrospermopsis raciborskii? Does a barcoding gap in the Planktothrix clade correspond to the described species?	1	2	life5010050_perova	1
+We understand why the reviewer would think that that would be beneficial. However, one has to keep in mind that we were not constructing a phylogenetic tree for all cyanobacteria but for single genera. In this case it is preferential to use a sister group that is more closely related to gain resolution within that group. e.g., Hedtke, S.M. ; Townsend, T.M. ; Hillis, D.M. Resolution of phylogenetic conflict in large data sets by increased taxon sampling. Syst. Biol. 2006, 55, 522–529. Moreover, for the sake of the barcoding analyses, the choice of the outgroup will not affect the results, given that it will not change the relationships towards the tips of the tree.	2	1	Why did you use Phormidium as an outgroup (Line 139)? Would not be more beneficial to use Gloeobacter or some other bacterium?	1	2	life5010050_perova	1
+The last paragraph of the discussion already included some comments on this issue. Now the paragraph has been expanded to provide a more detailed discussion on the possibility of using a DNA barcoding gap in cyanobacteria.	2	1	Is there any way to use barcoding gaps in a taxonomy of cyanobacteria? This might be added to discussion.	1	2	life5010050_perova	1
+We apologize for our carelessness in double checking the naming of the organisms and hope that we now eliminated all errors.	2	1	There are errors in nomenclature throughout the text, figures, and supplements, e.g. Leptolyngbya, Aphanizomenon, Arthrospira. You might check: http://www.cyanodb.cz/. It is an updated database of names of cyanobacteria.	1	2	life5010050_perova	1
+This review does not require any action on our part.	2	1	Overall a beautifully written, wonderfully presented, paper with clear concepts, methods and proposals that significantly contribute to original knowledge in the field.	1	2	life5010181_perova	1
+Here, the reviewer did not like our use of the particular words “novelty” and “innovation”. o Use of the word novelty − Instances of “chemical novelty” in the manuscript replaced by “emergent chemical behaviour”. − On Page 8, after Equation 8, we removed sentence: The emergence of bistability will serve as a proxy for the emergence of other chemical novelties in the vesicle reactor model. Replaced whole paragraph, to read: In particular, in these initial stages, we will focus on the emergence of bistability in the vesicle reactor model—a dynamical feature deducible directly from the number and stability of the fixed points present (i.e., two asymptotically stable points separated by an unstable saddle point). We also expect that more complicated dynamical regimes could also be present in the model, like multi-stability or global phase space features such as limit cycles giving rise to sustained oscillations. However, investigation of these regimes will be deferred to later work: for the time being, the “emergent chemical behavior” referred to in the title will be restricted to bistability. We think this is a clearer explanation, and also uses the word “regime” suggested by the reviewer. o Use of the word innovation We kept the 2 occurrences of the word “innovation” in the abstract and the introduction. An innovation is defined as a “new method, idea or product” in the dictionary. We use the word to refer to new emergent chemical behaviour that the whole vesicle system exhibits (e.g., expanded steady states), which did not exist before. We think the use of this word is acceptable.	2	1	The authors write: “In these initial stages, the emergence of bistability will serve as a proxy for the emergence of other chemical novelties in the vesicle reactor model.” The authors probably means that the existence of more than one stable regime could consent regime changes, but this situation do not indicates how these changes appear: in the article, the change is stimulated by means of a deliberated injection of chemical substances from outside. So, I think that the appearance of novelties requires a different order of considerations, whereas the bistability plays possibly the role of novelties amplification and consolidation (in particular conditions). The same observation can be done for the word “innovation”, used on the abstract and at Line 45. In order to avoid misunderstandings, I think that the authors should avoid these expressions, ad use the words “regime change”, or similar concepts.	1	2	life5010181_perova	1
+we added a new footnote, to explain our standing on this issue: Reference 36: In this work, concentrations outside the vesicle are set as system parameters. However, we make no commitment to the type of environment the vesicle is embedded in or how these concentrations are maintained. Our purpose is simply to show that bistability can exist in the model for certain sets of outside concentrations. Exploration of the model in explicit environments is deferred to future work.	2	1	Note 4 presents a very important issue: why the inner chemical environment is different from the external one, if the running chemical reactions are the same? The presence of particular features of the vesicle interior or of entrapped catalysts seem a too easy and ad hoc hypothesis. A useful reference proposing an explanation of this symmetry breaking (without requiring different conditions in internal and external environments) could be “R. Serra, M. Villani Mechanism for the formation of density gradients through semipermeable membranes Physical Review E 87, 2013”.	1	2	life5010181_perova	1
+Reference 11 has been added after Equation 10, to point the reader to where to find information about phi limits was first discussed. Repeating this information here would complicate the paper. The lines Phi = 2^(1/3) and 4^(1/3) in Figure 1d have now been explained.	2	1	How the authors derive the range [0.9, 101/3] for the so called “reduced surface” index? In Figure 1d, Lines 21/3 and 41/3 have some particular meanings, or are plotted mainly to ease the figure comprehension? Could the authors add some indications about the derivation of these upper limits?	1	2	life5010181_perova	1
+It is difficult to make a direct and meaningful comparison like this. Without a container, the reaction system has less parameters, and thus the sampled parameter space is smaller. The aim in the paper, was to simply show that encapsulating a Schlogl model which was bistable in bulk condition, seemed to destroy this bistability.	2	1	The authors write (case 1): “of the 5000 parameter set tested under constant surface area, 82% (4098) gave a single fixed point, 321 15.7% (785) gave two fixed points, and 2.3% (117) no fixed points.” How this statistics compare with a similar one, performed on a similar chemical situation without container? Could the authors add the results of the same within a not confined environment?	1	2	life5010181_perova	1
+The minor issues were fixed.	2	1	Adamala, K.; Szostak, J. ompetition betweenmodel protocells driven by an encapsulated catalyst. Nat. Chem. 2013, 5, 495–501. → “ompetition” should be: “Competition”.	1	2	life5010181_perova	1
+This immunoquantitation method used is described in detail in the reference given (Brown et al. 2008). The reviewer is directed in particular to the Supplementary Methods section. To allow readers to get a better understanding of the method in this manuscript we have revised this part of the Experimental Section, adding extra text to increase clarity concerning the method.	2	1	The data are interesting and worth publishing but… (1) I don’t understand how they calculated the protein molar ratios. In the methodology section we learn that they extracted the crude proteins and then performed western analyses using specific antibodies, followed by image analyses. I don’t understand how they converted the western data to read the amount of specific proteins, in fmol, in the various organisms as shown in Figure 2; and used for the calculations in the rest of the paper. (Why are the units provided again in Line 220?). Please provide the reader with the methodology used for the quantification.	1	2	life5010403_makarova	1
+Since submitting the manuscript, we have accumulated a set of 84 parallel measurements of steady state oxygen evolution and the functional content of PSII measured using flash yields (with a solid state optode) and simultaneous FRR chlorophyll fluorescence induction curves, from which we can extract e- PSII-1 s-1, for both Synechococcus and Prochlorococcus cultures. Consistent with the long literature history of such measurements (ex. Suggett et al. 2004, 2009), we observe a good correlation between the two measures of electron transport per PSII. Plotting the FRR estimate of e- PSII-1 s-1 versus the O2 evolution/PSII content estimate of e- PSII-1 s-1 gives a slope of 1.26 and an R2 of 0.58. We have added this information to the Materials and Methods to support our use of FRR estimates of electron transport per PSII. Furthermore, our estimates of ETRmax from FRR induction curves are independently validated by the close correlation between ETRmax and 1/tau, shown in Figure 4A, since 1/tau is derived from the rate constant for the decay of fluorescence after induction, and thus does not depend (computationally) upon our estimator for ETRmax.	2	1	PSII activity is derived from fluorescence measurement, as ETR. I am missing calibration with real measurements of PSII activity such as O2 evolution. You have got to show it for each organism you are examining otherwise it is worthless. There are many reports in the literature showing dramatic decline in fluorescence but hardly any change in O2 evolution.	1	2	life5010403_makarova	1
+This has been addressed with the addition of text in the abstract, introduction and discussion specifying that the cultures were grown under low light conditions.	2	1	The cells were grown under a relatively low light intensity and thus the data presented here only apply to these conditions. Please make sure the reader is aware of it.	1	2	life5010403_makarova	1
+We have removed the word “nondiazotrophic” as we agree that it is not relevant to the discussion.	2	1	Line 19 – I am not sure why stating the cyanobacteria are nondiazatrophic is relevant?	1	2	life5010403_makarova	1
+We mean that they numerically dominate and we have clarified this in the introduction.	2	1	Line 46 – (and throughout) be clear if you mean numerically dominate or dominate production – they are not always the same thing.	1	2	life5010403_makarova	1
+We have clarified this statement in the text. We refer to the nitrogen cost in the form of allocation to protein per pigment bound.	2	1	Introduction paragraph 2 – I am unsure we know that physbobilisomes require more resource than pcb proteins – this depends on the ratio of antenna to reaction centre? Please include references if R3 this has been shown. Also, not all Pcb proteins are constitutively expressed (e.g., see Bibby et al. Nature 2003).	1	2	life5010403_makarova	1
+We are referring to the number of RUBISCO active sites measured by immunoquantitation. We are not referring to measured RUBISCO activity. We use this expression to be clear that we are referring to RbcL subunits rather than oligomeric RUBISCO. We have added a parenthetic phrase to make this more clear.	2	1	Line 250 – This is estimated number of RUBISCO active sites – or is there evidence all the RUBSCIO you quantify is active?	1	2	life5010403_makarova	1
+We thank the reviewer for drawing this study to our attention. We have added a paragraph to the discussion to compare and contrast the Sukenik work with that presented here. The Sukenik data support a strong positive correlation between 1/tau and the RUBISCO to PSU ratio over a series of growth irradiances. While we also see a strong positive correlation between 1/tau and RUBISCO to PSII our results differ as the molar ratios of the components of the PSU differ significantly between the strains analyzed in the current work. This allows us to pinpoint the relationship of PSII to RUBISCO rather than other subunits of the PSU as the determinant of electron transport rate.	2	1	Figure 4 – The crunch of this paper is Figure 4e which show the Rubisco:PSII ratio is correlated to ETR. While I’m convinced in the presented data only three data-points are shown – I’m most convinced in that this same relationship is presented in in “Aquatic Photosynthesis (edition 2) Figure 7.9 – Falkowski” based on work of Sukenik (1986?). A greater discussion should be given in the text to how these datasets compare and what is significantly new in this dataset? References 35 and 36 are discussed in relation to the potential control of cytb6f on limiting electron transfer from PSII but more should be made of Falkowski’s observations which support this data.	1	2	life5010403_makarova	1
+We have added text to the end of the discussion to address this comment and the relevant reference has been added. Thank you for this suggestion.	2	1	Line 308 – The data presented is used to support potential cyclic electron flow around PSI – however other alternative terminal electron sinks could also be up-regulated – these concepts are discussed in a recent review by Milligan and Behrenfeld annual review 2013 – the implications of this should be discussed unless the authors can show specifically enhanced flow around PSI.	1	2	life5010403_makarova	1
+We have added a supplemental figure (Supplemental Figure #2) to show this calibrations curve. As all experiments presented here were performed under iron replete conditions, no iron starvation was performed.	4	1	“In line 178 onward we read about the calibration curve. Please presnt it in the paper. Once you apply stress such as iron starvation you should recalibrate.” R4 Response:	3	2	life5010403_makarova	1
+We have added a supplemental figure (Supplemental Figure #1) that shows the method applied with a sample blot, calibration curve and data analysis. This work required dozens of blots, each with its own standard curve, so it would not be practical to show all of the standard curves for each determination.	4	1	“Please present the calibration curves for the protein levels.” Response:	3	2	life5010403_makarova	1
+p. 7, Figure 3, formate formula has been changed to “HCOO-” from “COOH-”.	2	1	In Figure 3, formate formula should be better symbolized as HCOO- instead of COOH- as indicated.	1	2	life5010432_makarova	1
+p. 2, Lines 22–24; the data at the website cannot be posted as a supplementary figure because of copyright issues. I will ask them to revise the figure using English.	2	1	Lines 59–60: Instead of links, a figure (may be included in supplementary document) will be more helpful. The links lead to a non-English website, and also they may become outdated in year or two.	1	2	life5010432_makarova	1
+p. 2, Line 24; I confirmed that the following website link to the data of nitrite concentration is correct. “http://www.data.jma.go.jp/gmd/kaiyou/db/vessel_obs/hq/2006spr/137e/index_line.php?id=no2” 	2	1	Line 60: Link to NO2− concentrations shows temperature profiles.	1	2	life5010432_makarova	1
+p. 4, line 16; Six NitM proteins registered recently were added to Figure 1 and Figure 5.	2	1	Line 118: Was the analysis done on all FNT-proteins available at NCBI database? Please clarify.	1	2	life5010432_makarova	1
+p5, Figure 1; Figure 1 was reproduced by using the UPGMA clustering method instead of the NJ clustering method according to the suggestion of the reviewer, then, NitM from α-cyanobacteria and β-cyanobacteria form clearly distinct groups. “using the UPGMA (Unweighted Pair Group Method with Arithmetic mean) clustering method of ClustalX.” was added in the legend of Figure 1.	2	1	Line 40: Line 120: From the tree as it is, one can see that NitM from β-cyanobacteria form a distinct clade. However, NitM from α‐cyanobacteria are not grouped in one clade. It is possible that another outgroup sequence or different methods for alignment and tree construction will help. Line 122: What methods were used for creating alignment and phylogenetic tree (for example, neighbor-joining)?	1	2	life5010432_makarova	1
+Formate inhibition experiments of the NitM from CC9311, CC9605 and MIT9313 have not been carried out yet.	2	1	Was formate inhibition test also done on strains with nitM from CC9311, CC9605 and MIT9313?	1	2	life5010432_makarova	1
+We will consider the nitrite uptake experiments at environmentally relevant concentrations of nitrite (~50 nM) in the future.	2	1	It would be especially exciting to see the uptake of nitrite at environmentally relevant concentrations (less than 50 nM)? Considering the genetic constructs available, I was wondering if the authors have attempted such experiments and what were the results.	1	2	life5010432_makarova	1
+p. 7, Figure 3, formate formula has been changed to “HCOO-” from “COOH-”.	2	1	In Figure 3, formate formula should be better symbolized as HCOO- instead of COOH- as indicated.	1	2	life5010432_perova	1
+p. 2, Lines 22–24; the data at the website cannot be posted as a supplementary figure because of copyright issues. I will ask them to revise the figure using English.	2	1	Lines 59–60: Instead of links, a figure (may be included in supplementary document) will be more helpful. The links lead to a non-English website, and also they may become outdated in year or two.	1	2	life5010432_perova	1
+p. 2, Line 24; I confirmed that the following website link to the data of nitrite concentration is correct. “http://www.data.jma.go.jp/gmd/kaiyou/db/vessel_obs/hq/2006spr/137e/index_line.php?id=no2” 	2	1	Line 60: Link to NO2− concentrations shows temperature profiles.	1	2	life5010432_perova	1
+p. 4, line 16; Six NitM proteins registered recently were added to Figure 1 and Figure 5.	2	1	Line 118: Was the analysis done on all FNT-proteins available at NCBI database? Please clarify.	1	2	life5010432_perova	1
+p5, Figure 1; Figure 1 was reproduced by using the UPGMA clustering method instead of the NJ clustering method according to the suggestion of the reviewer, then, NitM from α-cyanobacteria and β-cyanobacteria form clearly distinct groups. “using the UPGMA (Unweighted Pair Group Method with Arithmetic mean) clustering method of ClustalX.” was added in the legend of Figure 1.	2	1	Line 40: Line 120: From the tree as it is, one can see that NitM from β-cyanobacteria form a distinct clade. However, NitM from α‐cyanobacteria are not grouped in one clade. It is possible that another outgroup sequence or different methods for alignment and tree construction will help. Line 122: What methods were used for creating alignment and phylogenetic tree (for example, neighbor-joining)?	1	2	life5010432_perova	1
+Formate inhibition experiments of the NitM from CC9311, CC9605 and MIT9313 have not been carried out yet.	2	1	Was formate inhibition test also done on strains with nitM from CC9311, CC9605 and MIT9313?	1	2	life5010432_perova	1
+We will consider the nitrite uptake experiments at environmentally relevant concentrations of nitrite (~50 nM) in the future.	2	1	It would be especially exciting to see the uptake of nitrite at environmentally relevant concentrations (less than 50 nM)? Considering the genetic constructs available, I was wondering if the authors have attempted such experiments and what were the results.	1	2	life5010432_perova	1
+Multi-alignment of concatenated protein (or DNA) segments is a genome-scale, but not whole-genome approach. Its applicability depends on the scope of the phylogenetic study. When dealing with not-too-distantly related species it may yield more or less useful result. However, in a study covering many phyla it is very difficult, if not impossible, to collect a common set of conserved proteins. Moreover, the concatenation method can never lead to very convincing conclusion, as give or take a few proteins may change the result. The phylogenomics people have noticed this problem, see, e.g., O. Jeffroy, H. Brinkman, F. Delsuc, H. Philippe (2008) Phylogenomics: the beginning of incongruence? Trends in Genetics, 22(4): 225–231. An example from the Bacteria domain is the relationship of the closely related Shigella and Escherichia coli strains. Concatenation of different number of genes led to different way of mixing-up of the two groups, but CVTree gave unambiguous separation of the strains as different species in the same genus Escherichia, see: G.-H. Zuo, Z. Xu, B.L. Hao (2013) Shigella strains are not clones of Escherichia coli but sister species in the genus Escherichia. Genomics Proteomics Bioinformatics, 11: 61–65. In order to carry out multi-alignment of concatenated sequences, a postdoc or well-trained PhD student equipped with the corresponding software is required. In contrast, with genome sequencing becoming a common practice in many labs it costs no additional work for a bench-microbiologist to get phylogenetic and taxonomic information by using a convenient and publically available tool such as the CVTree web serve. Well, we would be glad to see comparison of CVTree phylogeny with multi-alignment of concatenated proteins if anyone finds a way to do it for so many diverse phyla, but we do not consider it as a doable job.	2	1	I think the alignment-free methodology is interesting, however I would like to see a comparison with at least one regular alignment/treeing method, based on the same genomes the authors used, and not just a visual topological comparison with other published trees.	1	2	life5010949_makarova	1
+These points were discussed in the “Material and Method” section added at the suggestion of Reviewer 2. The following was copied from the manuscript: “Traditionally a newly generated phylogenetic tree is subject to statistical re-sampling tests such as bootstrap and jackknife. CVTree does not use sequence alignment. Consequently, there is no way to recognize informative or non-informative sites. Instead we take all the protein products encoded in a genome as a sampling pool for carrying out bootstrap or jackknife tests (citing our 2004 paper). Although it was very time-consuming, CVTrees did have well passed these tests (citing our 2010 paper). However, successfully passing statistical re-sampling tests only tells about the stability and self-consistency of the tree with respect to small variations of the input data. It is by far not a proof of objective correctness of the tree. Direct comparison of all branchings in a tree with an independent taxonomy at all ranks would provide such a proof, The 16S rRNA phylogeny cannot be verified by the Bergey's taxonomy, as the latter follows the former. However, agreement of branchings in CVTree with the Bergey's taxonomy would provide much stronger support to the tree as compared to statistical tests. This is the strategy we adopt for the CVTree approach.” “There are two aspects of a phylogenetic tree: the branching order (topology) and the branch lengths. Branching order is related to classification and branch length to evolution time. Calibration of branch lengths is always associated with the assumption that mutation rate R3 remains more or less a constant across all species represented in a tree, an assumption that cannot hold true in a large-scale phylogenetic study like the present one. Therefore, branching order in trees is of primary concern, whereas calibration of branch lengths makes less sense. Accordingly, all figures in this paper only show the branching scheme without indication of branch lengths and bootstrap values”.	2	1	As far as I can see, statistical support on the branches is missing, so I have no way of assessing if this branching order is valid. Bootstrapping or jackknifing are by no means the final word on the significance of branches, however an explanation as to how the user should assess the significance of braches would be good, i.e., branch length.	1	2	life5010949_makarova	1
+Yes, this is an apparent discrepancy of CVTree from 16S (and 23S) analysis for the given set of 179 archaeal genomes. However, in an on-going study of ours (not published yet) using a much larger data set this violation no longer shows up; both Korarchaeota and Crenarchaeota restore their phylum status. Taking into account the fact that both Korarchaeota and Thermofilaceae are represented by single species for the time being, their placement certainly requires further study with broader sampling of genomes.	2	1	The fact that Thermofilum is placed outside Crenarchaeota in Figures 3 and 4 is a little disturbing. I haven’t come across such a placement in other phylogenetic analyses of Archaea, for example in Brochier-Armanet et al 2008 Nat Rev Microb, or Rinke et al. 2013 Nature. I believe this needs to better explained in the manuscript, rather than just saying “this fact is noted”.	1	2	life5010949_makarova	1
+Highly degenerated genomes of many symbiont organisms tend to move around, in particular, to the baseline of a tree and thus distorts the overall structure of the tree. Therefore, it is better not to mix them with free-living organisms in a study. We rephrased the corresponding paragraph in the manuscript: “The nanosized archaean symbiont Nanoarchaeum equitans has a highly reduced genome (490,885 bp). It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota. We note that the monophyly of Euryarchaeota was also violated by Nanoarchaeum in some 16S rRNA trees, see, e.g., Figure 4 in a 2009 microbial survey as well as (c) and (d) in our Figure 3. It has been known that tiny genomes of endosymbiont microbes often tend to move towards baseline of a tree and distort the overall picture. In fact, we have suggested skipping such tiny genomes when studying bacterial phylogeny, see, e.g., (citing our 2010 paper) and a note in the home page of the CVTree Web Server. In the present case we may at most say that Nanoarchaeota probably makes a separate phylum, but its cutting into Euryarchaeota might be a side effect due to the tiny size of the highly reduced genome”.	2	1	I am also not very convinced with the placement of Nanoarchaeota. It seems like this phylum is moving around with the addition of new sequence data (for example in Rinke et al. Figure 2 tree, they are on an entirely different branch than Euryarchaeota). Though the authors also rightfully point out that the reduced genome size may have something to do with this placement.	1	2	life5010949_makarova	1
+Yes, there was certain disturbing effect of the tiny and lonely Nanoarchaeum genome, yet the Halobacteria is a very specific clade, forming a tightly connected group and moving around as a whole, mainly due to the biased acidity of their constituent amino acids. We anticipate that the relative placement of Halobacteria with respect to other groups may stabilize when more genomes are used to construct a tree.	2	1	The placement of Halobacteria (due to interfering Nanoarchaeota, I presume) is also a little disturbing. I would recommend that the authors provide a discussion of this. Especially with regards to other archaeal trees. For instance, in the tree of Armanet et al 2011 that the authors also refer to, the placement of Halobacteria with respect to Nanoarchaeota is very different.	1	2	life5010949_makarova	1
+We have A new “Material and Method” section has been added. Such issues as statistical resampling tests (bootstrap and jackknife), calibration of branch length, the meaning and choice of the peptide length K, etc. , were discussed in the new section. Figures 1 and 2 were combined to a new Figure 1; Figures 3 and 4 were combined to become a new Figure 2. Figure captions were made more detailed. The whole text was checked for language flaws and many places were rephrased.	2	1	In summary, I find the piece interesting, but parts of the discussion are rather weak, therefore I am suggesting a major revision. Another reason for major revision is the style that the manuscript is written. I am not a native speaker, but given that I had to read sentences several times, I suspect the manuscript can benefit from an English language check.	1	2	life5010949_makarova	1
+We thank the Reviewer for the detailed comments/suggestions given in the previous report and the suggestion of doing spelling-check this time. We have gone through the final manuscript carefully once more.	4	1	I found the revised version of this manuscript quite good, and I thank the authors for responding thoroughly to all my comments.	3	2	life5010949_makarova	1
+A “Material and Method” section has been added where the CVTree algorithm, the interactive tree-viewer, statistical resampling tests (bootstrap, jackknife), calibration of branch lengths, etc., were discussed in slightly more detail.	2	1	I would really appreciate a “Methods” part where the CVTree is explained shortly, and the tree-viewer is explained with more detail. The absence of branch lengths and bootstraps should be discussed here. Other technical aspects of the paper (e.g., sequence dataset), parameters, criteria ... all could be well organized in this part.	1	2	life5010949_makarova	1
+Yes, 16S rRNA phylogeny is quite stable and it almost defines the present taxonomy. We have given due credit for this. In general, CVTree does not challenge 16S rRNA analysis but complement it.	2	1	Archaeal phylogeny has already been studied in detail, with 16S and other marker genes, and with genomic approaches too. Some of the undersigning authors had already published on this before, although with smaller input datasets. Therefore, the fact that 16S topology is quite stable and comparable with other approaches is already known.	1	2	life5010949_makarova	1
+A robust phylogenetic tree comes with a fixed branching order of leaves. One looks at the leaf names and their taxonomic lineage and tries to map the latter to the branches. To this end we added the following paragraphs in the “Material and Method” section. “There are two aspects of a phylogenetic tree: the branching order (topology) and the branch lengths. Branching order is related to classification and branch length to evolution time. Calibration of branch lengths is always associated with the assumption that mutation rate remains more or less a constant across all species represented in a tree, an assumption that cannot hold true in a large-scale phylogenetic study like the present one. Therefore, branching order in trees is of primary concern, whereas calibration of branch lengths makes less sense. Accordingly, all figures in this paper only show the branching scheme without indication of branch lengths and bootstrap values.” “Branching order in a tree by itself does not bring about taxonomic ranks, e,g, class or order.	2	1	Taxonomists have traditionally circumscribed the high taxa (specially orders and classes) with great subjectivity, i.e., without well accepted criteria. In terms of phylogenetic trees one premise has always been clear, a taxon must be monophyletic. This principle has been used in the present work to reconsider the status of some high taxa. However, authors do not explain objective criteria to properly interpret the rank of the clades, which impedes making a profound evaluation of the archaeal classification. Therefore, although authors have strong tools and dataset, they just achieved a small revision of the high taxa which is, indeed, quite biased by the underlying 16S guidelines.	1	2	life5010949_makarova	1
+We have reorganized the manuscript mainly by adding a new “Material and Method” section where discussions on branch length, statistical resampling, meaning and choice of K, etc., were given. The original Figure 1 was deleted with some related points explained in the text accompanying the original Figure 2. All figure captions have been rewritten for clarity.	2	1	I have noted some lack of scientific rigor according to: many wrong taxonomic names and typos, scarce figure legends, few comments about the missing branch lengths or bootstraps (! ), redundancy in text and figures and fragments which are really difficult to understand. Authors should pay attention to language, explanations and text organization.	1	2	life5010949_makarova	1
+A few more sentences were added in the “Conclusion” regarding the power and achievement of the 16S rRNA analysis.	2	1	Authors shouldn't forget (particularly in conclusion) that the resolution power of 16S for high ranks (genus and above) is currently well accepted. And the number of non-redundant 16S entries available is much much larger than that of archaeal genomes. 16s data offer a much comprehensive view of the archaeal diversity, including deep branches.	1	2	life5010949_makarova	1
+We tried to rephrase the paragraph by changing, deleting, or adding a few words as follows: “In this paper we study Archaea phylogeny across many phyla. This is in contrast with phylogeny of species in a narrow range of taxa, e.g., that of vertebrates (a subphylum) or human versus close relatives (a few genera). Accordingly, the phylogeny should be compared with taxonomy at large, or, as Cavalier-Smith (citing cavalier-smith 2002) put it, with “megaclassificaton” of prokaryotes. Although in taxonomy the description of a newly discovered organism necessarily starts from the lower ranks, higher rank assignments are often incomplete or lacking. At present the ranks above class are not covered by the Bacteriological Code. The number of plausible microbial phyla may reach hundreds and archaeal ones are among the less studied. According to the 16S rRNA analysis, the major archaeal classes and their subordinate orders have been more or less delineated. Therefore, in order to carry out the aforementioned cross verification we make emphasis on higher ranks such as phyla, classes, and orders. A study using 179 Archaea genomes should provide a framework for further study of lower ranks.”	2	1	- L40 “Since at present (…) are not covered (...)” is a weak reason for choosing high ranks. I recommend to shortly summarize why high ranks are so important, and why do you choose order as the lowest considered rank.	1	2	life5010949_makarova	1
+Branching order in a tree is directly related to taxonomy, while branch lengths have more to do with evolution. For large-scale phylogenetic study across many phyla the former is more important than the calibration of branch lengths. The latter is based on the assumption that mutation rate is more or less constant. This assumption cannot hold when dealing with many phyla.	2	1	Part 3.1 The figure legends require more rigorous explanation. Would be interesting to remember that the branch lengths are not taken into account, or were omitted.	1	2	life5010949_makarova	1
+This is done in the newly added “Material and Method” section.	2	1	In Line 80—authors write some explanations to understand the tree figures. I would suggest to put this text before the first tree figure.	1	2	life5010949_makarova	1
+The original Figure 1 was deleted and a few words added to the legend of the original Figure 2, now the new Figure 1.	2	1	Figure 1 is a bit redundant. A short comment about the inclusion of that particular sequence into the Thermoplasmatales can be added into the legend of Figure 2.	1	2	life5010949_makarova	1
+Judging by the cluster labeled as Euryarchaeote{0+3} in Figure 2 Methanomassiliicoccus was not reclassified into Thermoplasmataceae but to an yet un-specified class.	2	1	However, the reclassification of Methanomassiliicoccus into Thermoplasmataceae needs more explanation. To be objective, authors should address the following question, why in the same family and not in another new family?	1	2	life5010949_makarova	1
+The sentence has been moved to the legend of Fig. 1 and slightly rephrased.	2	1	L106–107: maybe a bit inappropriate on that position. I would suggest to add a comment on the figure legend instead.	1	2	life5010949_makarova	1
+Done.	2	1	Part 3.2 L109–118: Summarize and move to introduction. Those sentences are of general importance for the topic and not specific to part 3.2.	1	2	life5010949_makarova	1
+the K issue is discussed in the newly added “Material and Method” section; so scattered mentioning of K has been deleted from the rest of text.	2	1	L119–123: if the K issue is relevant to understand the text then please add a proper explanation. If not, then keep it simple and avoid entering into the K issue (L122–123, L132, L135, L139, 183, and also remove this K = 6 from Figure 3.)	1	2	life5010949_makarova	1
+Done in the caption of Figure 2.	2	1	Add more explanations in legend of Figure3.	1	2	life5010949_makarova	1
+We have rewritten the paragraph as: “The new phylum Korarchaeota violates the monophyly of the phylum Crenarchaeota by drawing to itself the family Thermofilaceae. However, in an on-going study of ours (not published yet) using a much larger data set, this violation no longer shows up; both R8 Korarchaeota and Crenarchaeota restore their phylum status. Taking into account the fact that both Korarchaeota and Thermofilaceae are represented by single species for the time being, their placement certainly requires further study with broader sampling of genomes.”	2	1	L143: Sounds clearer if you avoid mixing class and phylum, for example: “The placement of phylum Korarchaeota, as a closest neighbor of family Thermofilaceae, violates the monophyly of phylum Crenarchaeota.” Response:	1	2	life5010949_makarova	1
+From the original Figures 3 and 4 only one has been kept and the legend rewritten. In fact, the whole paragraph changed to: “The newly proposed phylum Thaumarchaeota appears to be non-monophyletic as an outlying strain Candidatus Caldiarchaeum subterranum was assigned to this phylum according to the NCBI taxonomy. The NCBI assignment might reflect its position in some phylogenetic tree based on concatenated proteins, e.g., Figure 2 in […]. However, in the original paper reporting the discovery of this strain […] and in recent 16S rRNA studies, e.g., […], Candidatus Caldiarchaeum subterranum was proposed to make a new phylum Aigarchaeota. CVTrees support the introduction of this new phylum. A lineage modification of Candidatus Caldiarchaeum subterranum from Thaumarchaeota to Aigarchaeota would lead to a monophyletic Thaumarchaeota{7}.”	2	1	L146: No need to explain the 6+2 if it is properly explained in the figure legend. Figure 3 is redundant. I would recommend to avoid presenting different versions of the same tree; just the final tree is OK (use final/valid labels) and all important explanations in the text or legend. Perhaps the whole reasoning in Lines 142–180 is not so relevant for the current objective of comparing CVT, LTP, Bergeys? Or, perhaps, define this objective more clearly.	1	2	life5010949_makarova	1
+The problem of taxonomic placement of Aciduliprofundum is a good example to demonstrate how one extract information from CVTrees. In the Reysenbach et al. Nature 2006 paper it was taken as the first cultivated member of the DHEV2 (deep-sea hydrothermal euryarchaeate 2) clade based on a maximum-likelihood 16S rRNA tree. Unfortunately, all other 13 members of this clade were represented by 16S rRNA sequences only and no genome data are available so far. The NCBI taxonomy gave an incomplete lineage: Archaea; Euryrchaeota; unclassified Euryarchaeota; missing taxonomic assignment at the rank class and below. In order to make use of CVTree we must touch on the K-issue a little more. The alignment-free comparison of genomes in CVTree is implemented by counting the number of K-peptides in the protein products encoded in a genome followed by subtraction of a random background caused by neutral mutations. The peptide length K looks like a parameter, but it is actually not a parameter. Using a longer K emphasizes species-specificity, while a shorter K takes into account more common features with neighboring species. However, we never adjust K: a fixed K is used for all genomes to construct a tree, but one may construct a series of trees for K = 3, 4, 5, 6, 7, 8, … We have shown repeatedly that R9 K = 5 and 6 lead to best results in the sense of agreement with taxonomy, so usually only a K = 6 tree is given in publications. Let us look at a subtree, i.e., part of a tree, containing the organisms of interest. If the branching order in all trees built for different Ks turns out to be same, it would be a strong support to the branching order. In most cases the branching order varies with K: K = 3 and 4 make sense, K = 5 and 6 yield the best, K = 7 and 8 become slightly worse, etc. For too big a K, even if the closest strains remain grouped together the whole tree may tend to become a star-tree, i.e., every small clade stands in its own and their mutual placements become less meaningful. Therefore, inspection of trees for a range of K-values provides an additional dimension to evaluate the results. For Aciduprofumdum we have a stable pair (<C>Thermococci{18}, <G>Aciduprofumdum{2}) at K=3, 5, 6, 7. At K = 4 we have (<C>Thermococci{18}, (<G>Staphylothermus{2}, <G>Aciduprofundum{2})) In all these cases Thermoplasmata stands farther away from the above pair. However, at K = 8 and 9, when the overall tree picture has been largely distorted, Aciduliprofundum does stand closer to Thermaplasmata. Putting together all the above results we tend to consider the pair (<C>Thermococci{18}, <G>Aciduliprofundum{2}) as reflecting a more probable relation. Confined to the available data for the time being one may assign <G>Aciduliprofundum to <C>Thermococci, e.g., to denote the pair as <C>Thermococci{20}=(<F>Thermococcaceae{18}, <G>Aciduliprofundum{2}) leaving its family unclassified or assign it to a new family. Without further phenotypic and chemotaxonomic evidence it is better not to introduce new taxon names if the present naming scheme is capable to accommodate the leaves without conflict. This was why we wrote “this modification would hold as long as no new facts challenge it”. Anyway, taxonomy has always been a work in progress. One has to be prepared for modifications when new data appear. To make a long story short, we have rewritten the paragraph as: “The Candidatus genus Aciduliprofundum is considered a member of the DHEV2 (deap-sea hydrothermal vent euryarchaeotic 2) phylogenetic cluster. No taxonomic information was given in the original papers [55,56]. The NCBI Taxonomy did not provide definite lineage information for this taxon at the class, order, and family ranks. According to [55] the whole DHEV2 cluster was located close to Thermoplamatales in a maximum-likelihood analysis of 16S rRNA sequences. A similar placement was seen in [54] where a Bayesian tree of the archaeal domain based on concatenation of 57 ribosomal proteins put a lonely Aciduliprofundum next to Thermoplasmata. However, in CVTrees, constructed for all K-values from 3 to 9, Aciduliprofundum juxstaposes with the class Thermococci{18}. An observation in [56] that this organism shares a rare lipid structure with a few species from Thermococcales may hint on its possible association with the latter. If we temporarily presume a lineage <C>Thermococci<O>Unclassified<F>Unclassified<G>Aciduliprofundum… R10 one might have a monophyletic class <C>Thermococci{20}. Since none of the 13 DHEV2 members listed in [55] has a sequenced genome so far, CVTree cannot tell the placement of the DHEV2 cluster as a whole for the time being. It remains an open problem as whether DHEV2 is close to Thermoplasmata or to Thermococci, or a new class is needed to accommodate DHEV2.”	2	1	L156–166. If I understood right, there was good support for Candidatus Aciduliprofundum as part of a clade called DHEV2, which is a sister clade of Thermoplasmatales. However in the present work the authors intend here to reclassify Aciduliprofundum into family Thermococcaceae of Thermococcales. This needs further explanation. Since the new affiliation is quite in disagreement with previous observations, and this is not properly justified in the results/discussion, the final statement “this modification would hold as long as no new facts challenge it” seems unacceptable. In addition, why should Aciduliprofundum be regarded as member of Thermococcaceae and not as another distinct family?	1	2	life5010949_makarova	1
+No, we did not mean it.	2	1	L163: If I’m right the current observation actually does not support the previous work done by Brochier-Armanet.	1	2	life5010949_makarova	1
+We should first explain how these inappropriate names appeared. We have insisted to use the directory name at the NCBI FTP site as genome name. However, in November 2013 NCBI announced that they would not release genomes of different strains of the same species as before. In a period thereafter NCBI sometimes put several genomes in a directory and we had to extract the data and to assign a name from the “Source” line of the GenBank file. This caused some confusion. For example, as of February 27, 2015, a directory name at NCBI remained “archaeon_Mx1201_uid196597” and we had to change it to: Candidatus_Methanomethylophilus_alvus_Mx1201_uid196597 Now all “wrong names” as pointed out by the Reviewer no longer appear in figures. In the text we tried to refer to their names as complete as possible.	2	1	L167–173: The names are wrongly written (please check the original submission). Authors have to explain with more clarity, why is this clade of rank class. If that is the case, is it a single-order class? A single family order?	1	2	life5010949_makarova	1
+The genome name at NCBI FTP site is “halophilic_archaeon_DL31_uid72619”. The uid number was dropped when mentioned in the text. We put it back and capitalized the first letter to “Halophilic”, still an illegal genus name.	2	1	L174–180: I don’t understand the reasoning along this paragraph. In addition “haolphiic_archaeon_DL31” is not well written, please be careful when copying names from other source. Also, a similar question about the objectiveness for detecting high taxa: why not to create new family?	1	2	life5010949_makarova	1
+It is Figure 2 in the revised manuscript. We discussed it at some length.	2	1	Figure 4: the reclassifications must be clearly justified in the text.	1	2	life5010949_makarova	1
+No, organism cannot be published. Thanks for correcting our mistake.	2	1	L191: organisms can't be validly published, but their names.	1	2	life5010949_makarova	1
+The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system. Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being. Even if defined in the future, it must be lineage-dependent. For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla. In addition, monophyly is a guiding principle in comparing branching order with taxonomy. Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy. If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.” R6 One cannot tell the rank of a node/leaf in a tree by simply looking at it. A reference taxonomy is alwys needed. We put the following in the “Material and Method” section to explain it: “Branching order in a tree by itself does not bring about taxonomic ranks, e,g, class or order.	2	1	Part 3.3 L221–222: Sure, but authors do not provide explanations about how do they know that a clade in a tree is a family, an order, a class, etc. There is a lack of criteria to reclassify the leaves.	1	2	life5010949_makarova	1
+The phrase “3063 identical nucleotide positions” was copied from the caption of Figure 4 of the cited Nunoura et al. 2011 paper without much thinking. We simply deleted it.	2	1	L224: I don’t understand “3063 identical nucleotide positions”. Why identical?	1	2	life5010949_makarova	1
+The whole paragraph has been rewritten as: “The nanosized archaean symbiont Nanoarchaeum equitans has a highly reduced genome (490,885 bp [44]). We note that the monophyly of Euryarchaeota was also violated by Nanoarchaeum in some 16S rRNA trees, see, e.g., Figure 4 in a 2009 paper [61] as well as (c) and (d) in our Figure 4. In fact, we have suggested skipping such tiny genomes when studying bacterial phylogeny, see, e.g., [29] and a note in the home page of the CVTree Web Server [21]. In the present case we may at most say that Nanoarchaeota probably makes a separate phylum, but its cutting into Euryarchaeota might be a side effect due to the tiny size of the highly reduced genome.”	2	1	L239–244: hard to read, please rephrase.	1	2	life5010949_makarova	1
+As a method CVTree is independent of 16S rRNA analysis. First, it uses protein products in a genome instead of RNA segments in the genome. Second, it does not do sequence alignment. CVTree generates stable trees but cannot tell which branch corresponds to what taxon. Only after comparison with the existing classification and nomenclature one would be able to make connections with taxonomy. In this sense it does depend on 16S rRNA taxonomy. Anyway, CVTree does not challenge 16S rRNA analysis but makes it more convincing in most cases. The revealed discrepancies call for further study.	2	1	Conclusion I disagree that CVTree approach is independent of 16S, because authors are using the current accepted classification (which is mainly 16S-based) to validate the observed clades.	1	2	life5010949_makarova	1
+In fact, genomic approaches are more effective at species level and below due to their high resolution power. At high ranks CVTree may be more effective in the sense that it does not require additional work. Suffice it to put genomes in CVTree web server and the branches come out, then compare them with a reference taxonomy.	2	1	Why at higher ranks, genomic approaches are more effective? That needs more explanation. And authors should also consider the large benefits of 16S data availability, specially at high ranks (genus and above) where the 16S has good resolution.	1	2	life5010949_makarova	1
+Even “herenow” does not seem to be an correct English word; we changed it to “so far”.	2	1	L208: erenow → herenow	1	2	life5010949_makarova	1
+We changed “is in contrast with” to “is distinct from” and added a phrase “focusing on taxonomy of higher ranks” at the end of a sentence. Now the sentences read: “This is distinct from phylogeny of species in a narrow range of taxa, e.g., that of vertebrates (a subphylum) or human versus close relatives (a few genera). Accordingly, the phylogeny should be compared with taxonomy at large, or, as Cavalier-Smith \cite{cavalier-smith2002} put it,with “megaclassificaton” of prokaryotes, focusing on taxonomy of higher ranks.” -	4	1	L35-38: I don’t get well the sentences which start from “This is in contrast ... ”. Please can you specify a bit more?	3	2	life5010949_makarova	1
+done	4	1	L45: should provide → provides	3	2	life5010949_makarova	1
+Moved to the conclusion section and the first word “Though” replaced by “In addition, since”	4	1	L47–49: move to conclusions?	3	2	life5010949_makarova	1
+The whole paragraph has been deleted. This paragraph was added to the revised manuscript because one of the Reviewers asked “Does CVTree still require input genome data to be annotated to gene features, i.e., protein or CDS?” Well, this question reminds us that for many so-called “Permanent Draft” genomes it may be worthwhile returning to our early practice of using whole genome nucleotide sequences without distinguishing coding and non-coding segments. Although it did not lead to better results as compared with using translated protein products, but it is doable on un-annotated contigs. We will try this later.	4	1	L81–85: I think this paragraph is interrupting a bit the text flow. I suggest deletion.	3	2	life5010949_makarova	1
+We have deleted all what appeared within the curly brackets and kept only “monophyletic Thaumarchaeota” as suggested. In fact, we could not tell how these words appeared there; there was none in our draft manuscript.	4	1	L244–245: I think the text within {} deviates the attention. I suggest delete that part, ending sentence with “monophyletic Thaumarchaeota” is also ok.	3	2	life5010949_makarova	1
+Bending modulus is a material constant which does not depend on the size and shape of the object, but on the composition of the vesicle membrane and aqueous solution used (Lines 121–123 in the new version). We have inserted a definition of the membrane bending modulus (Lines 115–116) while the text regarding this issue extends between Lines 112–128.	2	1	"It is an interesting topic, however, the presentation seems not to be clear. It would be better to have a short but clear description about what ""bending elasticity modulus"" is and its relevance to biological functions."	1	2	life5021101_makarova	1
+As the referee suggested The Theory and Experimental section were organized into the Section “Materials and Methods”. Other sections were renumbered accordingly.	2	1	The theory section could be in the Method section with both principle and procedures to get kc values.	1	2	life5021101_makarova	1
+As it was explained in the text our experimental method is developed for application to almost spherical vesicles. Vesicles with diameters between 20 and 40 micrometers were considered, whose deviations from spherical shape (fluctuation of the radius) were small in comparison with the mean sphere radius. We added the information concerning the shape and the size of the studied objects and added pictures of the equatorial cross section of a fluctuating vesicle (new Figure 3), as seen under the phase contrast microscope.	2	1	However, not all results seem to be presented. For example, it is not clear what kind of images were recorded, which were used to calculate the kc values, what the sizes/shapes of the fluctuating vesicles looked like. How these observations would be different or similar to other types of vesicles.	1	2	life5021101_makarova	1
+We have rewritten the introductory paragraph (Lines 46–60).	2	1	The first sentence in the introduction section should be corrected as cell should not be considered to be “the building block of life”. It’s better to re-write the “basic motivation” for the study.	1	2	life5021101_makarova	1
+Thank you for the comment, there was a mistake. We have tidied and updated the references.	2	1	There are two reference lists, one for 1–31 and another from 1–34. There are repeated ones. It’s not clear why the case was.	1	2	life5021101_makarova	1
+We thank the reviewer for further comments and hope that we can clarify the issues as given below. We have made changes in the manuscript (marked red) and added one new reference (ref. [35]).	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.	3	2	life5021101_makarova	1
+In principal in order to obtain elastic constants you need to apply force and measure the deformation that this force causes. If we want to obtain the bending elasticity of lipid vesicle we need a force with a very small power. In the case of thermally induced shape fluctuations as a force we use the Brownian (thermal) motion of water molecules, bombarding the membrane. This force is stochastic (we do not know its instant value), but the mean value of it is proportional to the temperature. As a result of this stochastic bombardment the lipid vesicle deforms (changes its shape or fluctuates). We acquire a big amount of pictures of fluctuating vesicle in order to get the mean value of the deformation that our force causes. To extract mean we need to have stationary conditions over the time, so the mean is taken, that is why it is really important to have constant temperature throughout the experiment.	4	1	If only measured at one temperature, how would it be possible to draw a conclusion with “thermally induced shape fluctuations”?	3	2	life5021101_makarova	1
+Using our experimental system we can measure bending elasticity also at higher temperatures (up to approximately 40 degrees C).	4	1	Is it possible to do the measurement at elevated temperatures?	3	2	life5021101_makarova	1
+The bending elasticity modulus depends on the temperature below and near the phase transition temperature, but far above the phase transition temperature (this is the case in our experiment) the bending elasticity modulus is practically constant. See: Temperature and Chain Length Effects on Bending Elasticity of Phosphatidylcholine Bilayers. Fernandez-Puente, I. Bivas, M. D. Mitov and P. Meleard, Europhys. Lett., 28, 181 (1994). We have added this text to the Discussion (Lines 234–237) and also added the reference to the reference list (ref.	4	1	Would kc be temperature dependent? If yes, what would be the valid range?	3	2	life5021101_makarova	1
+It is not a problem for us to make pictures of a fluctuating vesicle at different temperatures, but these photos would not be related to the manuscript.	4	1	The photo added merely shows the shape, but is it possible to show a few time related changes of the shape at different temperatures?	3	2	life5021101_makarova	1
+We thank the referee for pointing to negligent mistakes in numbering. We have corrected wrong numbering of subsections.	4	1	The numbering from Lines 306–307 may not be correct as there are two 2.2.	3	2	life5021101_makarova	1
+As the referee suggested, we have added data for the bending elasticity modulus of another lipid type—POPC lipid membrane. Also, we have added a new figure (Figure 3).	2	1	In addition, the number of tables and figures are too low for article. If could, the author should add other experimental data.	1	2	life5021101_makarova	1
+A number of improvements of the experimental procedure for the thermally induced shape fluctuation method were used in this study: the stroboscopic illumination was improved on several steps to achieve a better experimental conditions; a thermostatic stage was used to set and control a constant temperature, the analysing procedure was improved by adding strict objective criteria for qualification of the vesicle as a whole as well as for acceptance or rejection of a given contour of the sequence of recorded images and the white noise contribution to the amplitudes of thermal shape fluctuations was R3 evaluated and taken into account. We have given short description of the improvements made and for each of them cited the corresponding articles for detailed explanation. (Lines 181–184; 187–193).	2	1	In Page 1, Lines 27–28, the author described “the experimental set-up was improved …”. However, the authors have already reported the measurements of bending elasticity using stroboscopic illumination in author’s previous papers (such as Genova et al., J. Optoelectro. Adv. Mat., 2005). Describe how the experimental set-up was improved in this paper more precisely.	1	2	life5021101_makarova	1
+SOPC was chosen as both lipid types have transition temperature far below the temperature at experiments. In the revised version, we measured the bending modulus for another lipid type (POPC), measured and analysed by the same method for the same experimental conditions.	2	1	As a comparison of archaeal lipids, SOPC was used in this study. SOPC contains an unsaturated bond in its alkyl chains and its transition temperature seems to be quite low compared with archaeal lipids. What is the reason why the authors selected the SOPC for the representative of eukaryotic lipids?	1	2	life5021101_makarova	1
+We have corrected the wrong number.	2	1	In page 4, between line 108 and 109, this equation should be numbered as (6).	1	2	life5021101_makarova	1
+The aim of the present study was to obtain the bending elasticity modulus of the archaeal membrane at fixed temperature (27 degrees C) far above the phase transition temperature. We have not measured the temperature dependence of the bending constant.	2	1	Describe the temperature condition for measurement of fluctuation of GVs. Are there any effects of temperature on the fluctuation of GVs?	1	2	life5021101_makarova	1
+In this work we present the bending elasticity modulus of archaeal lipid membrane in pure water environment. We added the details concerning the water purification (Lines 173–175). In other works the influence of different admixtures in the aqueous solution around the membrane was studied and the values depending on the type and the concentration of it for given lipid were reported. As the referee suggested we added such information with the corresponding references in the introduction section (Lines 57–60).	2	1	In this paper, bending elasticity modulus was measured in pure water. Are there any effects of ionic strength or composition of solution on the fluctuation of GVs?	1	2	life5021101_makarova	1
+As I understand the referee had no further comments.	4	1	The manuscript was thoroughly revised according to reviewer’s suggestions and comments. Now this revised manuscript can be acceptable for publication.	3	2	life5021101_makarova	1
+Bending modulus is a material constant which does not depend on the size and shape of the object, but on the composition of the vesicle membrane and aqueous solution used (Lines 121–123 in the new version). We have inserted a definition of the membrane bending modulus (Lines 115–116) while the text regarding this issue extends between Lines 112–128.	2	1	"It would be better to have a short but clear description about what ""bending elasticity modulus"" is and its relevance to biological functions."	1	2	life5021101_perova	1
+As the referee suggested The Theory and Experimental section were organized into the Section “Materials and Methods”. Other sections were renumbered accordingly.	2	1	The theory section could be in the Method section with both principle and procedures to get kc values.	1	2	life5021101_perova	1
+As it was explained in the text our experimental method is developed for application to almost spherical vesicles. Vesicles with diameters between 20 and 40 micrometers were considered, whose deviations from spherical shape (fluctuation of the radius) were small in comparison with the mean sphere radius. We added the information concerning the shape and the size of the studied objects and added pictures of the equatorial cross section of a fluctuating vesicle (new Figure 3), as seen under the phase contrast microscope.	2	1	However, not all results seem to be presented. For example, it is not clear what kind of images were recorded, which were used to calculate the kc values, what the sizes/shapes of the fluctuating vesicles looked like. How these observations would be different or similar to other types of vesicles.	1	2	life5021101_perova	1
+It would also be helpful to clarify/expand the description about the drug delivery system using the archaeal vesicle. The first sentence in the introduction section should be corrected as cell should not be considered to be “the building block of life”. It’s better to re-write the “basic motivation” for the study.	2	1	We have rewritten the introductory paragraph (Lines 46–60).	1	2	life5021101_perova	1
+Thank you for the comment, there was a mistake. We have tidied and updated the references.	2	1	There are two reference lists, one for 1–31 and another from 1–34. There are repeated ones. It’s not clear why the case was.	1	2	life5021101_perova	1
+We thank the reviewer for further comments and hope that we can clarify the issues as given below. We have made changes in the manuscript (marked red) and added one new reference (ref. [35]).	4	1	It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones. The observed fluctuations of the membrane do not destabilize it. However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.	3	2	life5021101_perova	1
+In principal in order to obtain elastic constants you need to apply force and measure the deformation that this force causes. If we want to obtain the bending elasticity of lipid vesicle we need a force with a very small power. In the case of thermally induced shape fluctuations as a force we use the Brownian (thermal) motion of water molecules, bombarding the membrane. This force is stochastic (we do not know its instant value), but the mean value of it is proportional to the temperature. As a result of this stochastic bombardment the lipid vesicle deforms (changes its shape or fluctuates). We acquire a big amount of pictures of fluctuating vesicle in order to get the mean value of the deformation that our force causes. To extract mean we need to have stationary conditions over the time, so the mean is taken, that is why it is really important to have constant temperature throughout the experiment.	4	1	If only measured at one temperature, how would it be possible to draw a conclusion with “thermally induced shape fluctuations”?	3	2	life5021101_perova	1
+Using our experimental system we can measure bending elasticity also at higher temperatures (up to approximately 40 degrees C).	4	1	Is it possible to do the measurement at elevated temperatures?	3	2	life5021101_perova	1
+The bending elasticity modulus depends on the temperature below and near the phase transition temperature, but far above the phase transition temperature (this is the case in our experiment) the bending elasticity modulus is practically constant. See: Temperature and Chain Length Effects on Bending Elasticity of Phosphatidylcholine Bilayers. Fernandez-Puente, I. Bivas, M. D. Mitov and P. Meleard, Europhys. Lett., 28, 181 (1994). We have added this text to the Discussion (Lines 234–237) and also added the reference to the reference list (ref.	4	1	Would kc be temperature dependent? If yes, what would be the valid range?	3	2	life5021101_perova	1
+It is not a problem for us to make pictures of a fluctuating vesicle at different temperatures, but these photos would not be related to the manuscript.	4	1	The photo added merely shows the shape, but is it possible to show a few time related changes of the shape at different temperatures?	3	2	life5021101_perova	1
+We thank the referee for pointing to negligent mistakes in numbering. We have corrected wrong numbering of subsections.	4	1	The numbering from Lines 306–307 may not be correct as there are two 2.2.	3	2	life5021101_perova	1
+As the referee suggested, we have added data for the bending elasticity modulus of another lipid type—POPC lipid membrane. Also, we have added a new figure (Figure 3).	2	1	This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids. Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs. According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs. The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers. In addition, the number of tables and figures are too low for article. If could, the author should add other experimental data.	1	2	life5021101_perova	1
+A number of improvements of the experimental procedure for the thermally induced shape fluctuation method were used in this study: the stroboscopic illumination was improved on several steps to achieve a better experimental conditions; a thermostatic stage was used to set and control a constant temperature, the analysing procedure was improved by adding strict objective criteria for qualification of the vesicle as a whole as well as for acceptance or rejection of a given contour of the sequence of recorded images and the white noise contribution to the amplitudes of thermal shape fluctuations was R3 evaluated and taken into account. We have given short description of the improvements made and for each of them cited the corresponding articles for detailed explanation. (Lines 181–184; 187–193).	2	1	I also have questions and comments for acceptance. The list of comments is as follows. In Page 1, Lines 27–28, the author described “the experimental set-up was improved …”. However, the authors have already reported the measurements of bending elasticity using stroboscopic illumination in author’s previous papers (such as Genova et al., J. Optoelectro. Adv. Mat., 2005). Describe how the experimental set-up was improved in this paper more precisely.	1	2	life5021101_perova	1
+SOPC was chosen as both lipid types have transition temperature far below the temperature at experiments. In the revised version, we measured the bending modulus for another lipid type (POPC), measured and analysed by the same method for the same experimental conditions.	2	1	As a comparison of archaeal lipids, SOPC was used in this study. SOPC contains an unsaturated bond in its alkyl chains and its transition temperature seems to be quite low compared with archaeal lipids. What is the reason why the authors selected the SOPC for the representative of eukaryotic lipids?	1	2	life5021101_perova	1
+We have corrected the wrong number.	2	1	In page 4, between line 108 and 109, this equation should be numbered as (6).	1	2	life5021101_perova	1
+The aim of the present study was to obtain the bending elasticity modulus of the archaeal membrane at fixed temperature (27 degrees C) far above the phase transition temperature. We have not measured the temperature dependence of the bending constant.	2	1	Describe the temperature condition for measurement of fluctuation of GVs. Are there any effects of temperature on the fluctuation of GVs?	1	2	life5021101_perova	1
+In this work we present the bending elasticity modulus of archaeal lipid membrane in pure water environment. We added the details concerning the water purification (Lines 173–175). In other works the influence of different admixtures in the aqueous solution around the membrane was studied and the values depending on the type and the concentration of it for given lipid were reported. As the referee suggested we added such information with the corresponding references in the introduction section (Lines 57–60).	2	1	In this paper, bending elasticity modulus was measured in pure water. Are there any effects of ionic strength or composition of solution on the fluctuation of GVs?	1	2	life5021101_perova	1
+As I understand the referee had no further comments.	4	1	The manuscript was thoroughly revised according to reviewer’s suggestions and comments. Now this revised manuscript can be acceptable for publication.	3	2	life5021101_perova	1
+We have rephrased the four paragraphs where the referee found that the described examples are not always easy to follow.	2	1	However, being experts in their field the authors might not be aware that for readers less familiar with the metabolism/physiology of archaea, the examples are not always easy to follow, such as examples given on p. 7 starting line 196 (The main difference here is between a ferredoxin-dependent and/or a NAD-dependent decarboxylation of pyruvate into acetyl-CoA?/A few lines later when I read that “halophilic archaea do not contain the coenzyme methanopterin”, I had to go back and reread the preceding sentences to get an idea of the argument. ), p. 10 starting Line 299, and p. 12 starting Line 354.	1	2	life5021427_makarova	1
+We have added a short “outlook-type” paragraph towards the end of the conclusions. In this paragraph we mention three approaches that will facilitate gene discovery, among them large-scale screening techniques as exemplified by the insertion mutant library pointed out by the referee.	2	1	There is now available a great resource for the gene discovery in Haloferax volcanii (Kiljunen et al 2014: BMC Biology). A comprehensive insertion mutant library has been generated for this model archaeon in order to facilitate gene discovery. This should be mentitioned and discussed in the text; i.e., how this resource could be used to assign functions for genes.	1	2	life5021427_makarova	1
+We have rephrased the four paragraphs where the referee found that the described examples are not always easy to follow.	2	1	Giving specific examples is certainly a good idea. However, being experts in their field the authors might not be aware that for readers less familiar with the metabolism/physiology of archaea, the examples are not always easy to follow, such as examples given on p. 7 starting line 196 (The main difference here is between a ferredoxin-dependent and/or a NAD-dependent decarboxylation of pyruvate into acetyl-CoA?/A few lines later when I read that “halophilic archaea do not contain the coenzyme methanopterin”, I had to go back and reread the preceding sentences to get an idea of the argument. ), p. 10 starting Line 299, and p. 12 starting Line 354.	1	2	life5021427_perova	1
+We have added a short “outlook-type” paragraph towards the end of the conclusions. In this paragraph we mention three approaches that will facilitate gene discovery, among them large-scale screening techniques as exemplified by the insertion mutant library pointed out by the referee.	2	1	There is now available a great resource for the gene discovery in Haloferax volcanii (Kiljunen et al 2014: BMC Biology). A comprehensive insertion mutant library has been generated for this model archaeon in order to facilitate gene discovery. This should be mentitioned and discussed in the text; i.e., how this resource could be used to assign functions for genes.	1	2	life5021427_perova	1
+The authors would like to thank for this helpful comment. We changed the subsection as described below. With stating that argon-nitrogen mixtures mainly emit high amounts of radiation, the authors want to emphasize, that the sterilization efficiency is based on radiation rather than interaction of radicals with the sample. In low-pressure argon-nitrogen plasmas, the only significant possible radical is atomic nitrogen. As atomic nitrogen is not known for high etching efficiency towards organic materials, the main reason for inactivation of D. radiodurans is based on radiation effects. Furthermore, Argon-Nitrogen plasmas emit high amounts of radiation compared to Argon-Oxygen plasmas, as shown for example in [5]. The data in [5] is not comparable in absolute values, as the system was changed afterwards. Absolutely calibrated spectra were measured and have been submitted in another publication. The authors think that estimating the inactivation efficiency towards D. radiodurans based on the UV dose cannot be performed on a reliable bases due to the following reasons: First, the inactivation efficiency towards spores is not only depending on the overall dose of the emitted radiation, but especially on the dose per wavelength. This has been demonstrated by Munakata et al. for B. subtilis spores (Munakata et al., Photochemistry and Photobiology, Blackwell Publishing Ltd, 1991, 54, 761-768). To the authors’ knowledge, there is no publication measuring the wavelength depending inactivation efficiency of B. radiodurans in the VUV. Second, the samples are mixed with Mars Analog Soil and most of the samples are covered with it. This leads to a very inhomogeneous exposure of the samples which can only be estimated arbitrarily. To address the reviewer’s comments, subsection 2.3 has been modified as follows: An argon-nitrogen mixture (100:5 sccm) was used with continuous power of 500 W, as it emits significantly higher amounts of radiation in the bactericidal wavelength region from 100 nm to 400 nm compared to an argon-oxygen mixture (100:5 sccm) [5]. The absolute values presented in [5] are not applicable to the results shown here, due to modification of the plasma system concerning the power coupling. Nevertheless, the significantly higher emission of the argon-nitrogen mixture is shown. Absolute values of the radiation dose in the range from 100 nm to 400 nm of the system configuration used in this study were measured and submitted [6]. A possible radical that can interact with the samples is atomic nitrogen. As atomic nitrogen is not known for etching of organic materials, we assume that inactivation in the argon-nitrogen mixture is based on radiation effects. [6] Raguse, M.; Fiebrandt, M.; Denis, B.; Stapelmann, K.; Eichenberger, P.; Driks, A.; Eaton, P.; Awakowicz, P.; Moeller, R. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low pressure plasma sterilization. Submitted to Journal of Physics D: Applied Physics.	2	1	The plasma conditions are not described sufficiently. It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are. The term amount is not applicable here. With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.	1	2	life6020022_makarova	1
+The authors thank the referee for this comment that demonstrates that the authors need to clarify. However, to the authors it is not clear which potentially inactivating components of the plasma are not specified, as radicals and radiation are mentioned and the temperature is kept below 80°C to prevent inactivation effects due to heating. As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV. Thus, sputtering of the sample due to ion bombardment can be neglected. Low-pressure plasmas dissociate molecular gases up to several percent. Thus, the authors wanted to emphasize, that the atomic oxygen density and therewith the atomic oxygen flux to the samples is in the order of a few percent of the overall gas flux. Quantitative and even qualitative analysis of atomic oxygen densities by optical emission spectroscopy is not an easy task as several parameters like gas temperature, electron density and electron temperature are necessary. This is even more challenging in pulsed mode, as the plasma does not achieve a steady state due to switching it on and off to prevent intensive heating of the plasma. Determination of the gas densities and plasma parameters is currently addressed in extensive analysis of plasmas with different gas compositions but not finished and published yet. Nevertheless, the authors’ state, that a relevant amount of atomic oxygen species is produced in the low pressure argon-oxygen plasma as the minimum energy for dissociation of molecular oxygen is around 5 eV. The ionization energy for argon is 16 eV and 13 eV for molecular oxygen. Thus, sufficient electrons with enough energy to dissociate molecular oxygen will be present in the plasma for dissociation up to a few percent. In the authors’ view, estimating the mean free path of the atomic oxygen in the plasma is not necessary, as recombination of radicals in low pressure plasmas in the regime of a few Pascal takes place at the surfaces. Thus, any radical formed will be lost at the walls and not on its way to a surface. Estimating the mean free path of the radical in the Mars Analog Soil can only be performed on an arbitrary basis as the sticking and recombination coefficients of atomic oxygen are not known for the Mars Analog Soil. Thus, any calculation of the mean free path of the radical is based on estimated data and could only be verified by experiments. To address the reviewer’s comments, subsection 2.3 has been modified as follows: Only JSC Mars-1A analog soil was used in the second trial. In this case, an argon-oxygen mixture (100:5 sccm) was used as it produces reactive oxygen species due to dissociation of molecular oxygen in the plasma. Since radiation-based sterilization is faster than sterilization due to oxidation, the treatment time was increased. As this leads to heating of the sample above 80 °C in the continuous plasma, we used a pulsed mode, switching the plasma on and off at a frequency of 1 kHz and a duty cycle of 10 %. To account for this fact, the treatment time was increased to 45 min, yielding a plasma exposure time of 4.5 min. Furthermore, the power was increased to 1500 W, yielding a mean power of 150 W, as it is only applied 10 % of the time. The plasma conditions are not matched to each other as the main objective was to observe to what degree the cells of D. radiodurans were affected by radiation or reactive species and not which component is more efficient. Thus, sputtering of the sample due to ion bombardment can be neglected as possible inactivation mechanism.	2	1	The other potentially inactivating components of the plasma are not specified in the manuscript. For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description. Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.	1	2	life6020022_makarova	1
+The grain size distributions of the three Mars regolith simulants have been added to the text in form of two additional tables (table 2: P-MRS and S-MRS; and table 3: JSC Mars-1A), as well as a new citation for the grain size distribution of JSC Mars-1A [8].	2	1	Three different Mars regolith simulants were used. The mineralogical composition is given in table, but not the grain size distribution. Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.	1	2	life6020022_makarova	1
+To assess the shading effect of the regolith is difficult. As described we used a very thin layer of regolith particles, but did not measure them according to their shading effect but according to the weight, which was the same for all types of regolith. This experiment was meant as a test of the sterilization method adjusted to our purposes, as described in the introduction section. Therefore, we did not plan it according to the maximal theoretically achievable sterilization effect, but to design a life detection mission to Mars. [8] Wan, L., Wendner, R., & Cusatis, G. A Novel Material for In Situ Construction on Mars: Experiments and Numerical Simulations. 2016, arXiv preprint arXiv:1512.05461	2	1	The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.	1	2	life6020022_makarova	1
+The title has been changed.	2	1	First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_makarova	1
+In the introduction we already described the problem of the destructiveness of heat sterilization like autoclaving on the organic contents of a soil, which was our motivation to test an alternative method such as low temperature plasma sterilization. D. radiodurans was used as test organism because of its resistance towards UV radiation, which is a part of the sterilizing effect of plasma sterilization. The appropriate passages in the introduction section have been edited.	2	1	The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).	1	2	life6020022_makarova	1
+Since we did not intend to test the influences of the Martian Analog soils on D. radiodurans, but the sterilizing effect of the low temperature plasma treatment on cells intermixed with soil, we did not make a control to determine the effects of the Mars analog soils. The starting values of cell numbers were estimated on samples prepared the same way as the samples for the plasma sterilization (intermixed with soils and desiccated as described in the method section). They were incubated on TYG agar plates like the plasma samples after the treatment. This explanation was missing in the text and has now been added to the appropriate passages in the method section (Chapter 2.2.) for a better understanding.	2	1	In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.	1	2	life6020022_makarova	1
+The names are written in italics now, the mistake must have happened during the editing of the text.	2	1	Lines 31-32: Bacillus subtilis and B.pumilus should be written in italic.	1	2	life6020022_makarova	1
+Done, thank you.	2	1	Line 38: Deinococcus radiodurans should be written in italic. While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.	1	2	life6020022_makarova	1
+Yes, we just used one medium (TYG), so we changed it to ‘medium’.	2	1	Line 41: Medium instead of media. If I understood correctly, you used only one medium.	1	2	life6020022_makarova	1
+The writing has been changed accordingly.	2	1	Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Reply:	1	2	life6020022_makarova	1
+Since the bottles containing the Martian Analog soils were autoclaved before use, they should be sterile. We also made a separate test to check for sterility after autoclaving the Martian Analog soils, which has been negative (meaning no contamination could be detected via plating of the autoclaved JSC Mars-1A regolith on TYG medium).	2	1	Line 57: Are you sure that the Martian Analog soils were sterile before using?	1	2	life6020022_makarova	1
+An explanation has been added to the text passage.	2	1	Line 78: Please explain in the text what is “a vacuum treatment control”.	1	2	life6020022_makarova	1
+The differences might be caused due to the finer grain size of the other regolith used - particularly P-MRS, which possibly increased the shielding effect. This has been added to the discussion section.	2	1	Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?	1	2	life6020022_makarova	1
+The authors would like to thank for this helpful comment. We changed the subsection as described below. With stating that argon-nitrogen mixtures mainly emit high amounts of radiation, the authors want to emphasize, that the sterilization efficiency is based on radiation rather than interaction of radicals with the sample. In low-pressure argon-nitrogen plasmas, the only significant possible radical is atomic nitrogen. As atomic nitrogen is not known for high etching efficiency towards organic materials, the main reason for inactivation of D. radiodurans is based on radiation effects. Furthermore, Argon-Nitrogen plasmas emit high amounts of radiation compared to Argon-Oxygen plasmas, as shown for example in [5]. The data in [5] is not comparable in absolute values, as the system was changed afterwards. Absolutely calibrated spectra were measured and have been submitted in another publication. The authors think that estimating the inactivation efficiency towards D. radiodurans based on the UV dose cannot be performed on a reliable bases due to the following reasons: First, the inactivation efficiency towards spores is not only depending on the overall dose of the emitted radiation, but especially on the dose per wavelength. This has been demonstrated by Munakata et al. for B. subtilis spores (Munakata et al., Photochemistry and Photobiology, Blackwell Publishing Ltd, 1991, 54, 761-768). To the authors’ knowledge, there is no publication measuring the wavelength depending inactivation efficiency of B. radiodurans in the VUV. Second, the samples are mixed with Mars Analog Soil and most of the samples are covered with it. This leads to a very inhomogeneous exposure of the samples which can only be estimated arbitrarily. To address the reviewer’s comments, subsection 2.3 has been modified as follows: An argon-nitrogen mixture (100:5 sccm) was used with continuous power of 500 W, as it emits significantly higher amounts of radiation in the bactericidal wavelength region from 100 nm to 400 nm compared to an argon-oxygen mixture (100:5 sccm) [5]. The absolute values presented in [5] are not applicable to the results shown here, due to modification of the plasma system concerning the power coupling. Nevertheless, the significantly higher emission of the argon-nitrogen mixture is shown. Absolute values of the radiation dose in the range from 100 nm to 400 nm of the system configuration used in this study were measured and submitted [6]. A possible radical that can interact with the samples is atomic nitrogen. As atomic nitrogen is not known for etching of organic materials, we assume that inactivation in the argon-nitrogen mixture is based on radiation effects. [6] Raguse, M.; Fiebrandt, M.; Denis, B.; Stapelmann, K.; Eichenberger, P.; Driks, A.; Eaton, P.; Awakowicz, P.; Moeller, R. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low pressure plasma sterilization. Submitted to Journal of Physics D: Applied Physics.	2	1	The plasma conditions are not described sufficiently. It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are. The term amount is not applicable here. With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.	1	2	life6020022_perova	1
+The authors thank the referee for this comment that demonstrates that the authors need to clarify. However, to the authors it is not clear which potentially inactivating components of the plasma are not specified, as radicals and radiation are mentioned and the temperature is kept below 80°C to prevent inactivation effects due to heating. As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV. Thus, sputtering of the sample due to ion bombardment can be neglected. Low-pressure plasmas dissociate molecular gases up to several percent. Thus, the authors wanted to emphasize, that the atomic oxygen density and therewith the atomic oxygen flux to the samples is in the order of a few percent of the overall gas flux. Quantitative and even qualitative analysis of atomic oxygen densities by optical emission spectroscopy is not an easy task as several parameters like gas temperature, electron density and electron temperature are necessary. This is even more challenging in pulsed mode, as the plasma does not achieve a steady state due to switching it on and off to prevent intensive heating of the plasma. Determination of the gas densities and plasma parameters is currently addressed in extensive analysis of plasmas with different gas compositions but not finished and published yet. Nevertheless, the authors’ state, that a relevant amount of atomic oxygen species is produced in the low pressure argon-oxygen plasma as the minimum energy for dissociation of molecular oxygen is around 5 eV. The ionization energy for argon is 16 eV and 13 eV for molecular oxygen. Thus, sufficient electrons with enough energy to dissociate molecular oxygen will be present in the plasma for dissociation up to a few percent. In the authors’ view, estimating the mean free path of the atomic oxygen in the plasma is not necessary, as recombination of radicals in low pressure plasmas in the regime of a few Pascal takes place at the surfaces. Thus, any radical formed will be lost at the walls and not on its way to a surface. Estimating the mean free path of the radical in the Mars Analog Soil can only be performed on an arbitrary basis as the sticking and recombination coefficients of atomic oxygen are not known for the Mars Analog Soil. Thus, any calculation of the mean free path of the radical is based on estimated data and could only be verified by experiments. To address the reviewer’s comments, subsection 2.3 has been modified as follows: Only JSC Mars-1A analog soil was used in the second trial. In this case, an argon-oxygen mixture (100:5 sccm) was used as it produces reactive oxygen species due to dissociation of molecular oxygen in the plasma. Since radiation-based sterilization is faster than sterilization due to oxidation, the treatment time was increased. As this leads to heating of the sample above 80 °C in the continuous plasma, we used a pulsed mode, switching the plasma on and off at a frequency of 1 kHz and a duty cycle of 10 %. To account for this fact, the treatment time was increased to 45 min, yielding a plasma exposure time of 4.5 min. Furthermore, the power was increased to 1500 W, yielding a mean power of 150 W, as it is only applied 10 % of the time. The plasma conditions are not matched to each other as the main objective was to observe to what degree the cells of D. radiodurans were affected by radiation or reactive species and not which component is more efficient. Thus, sputtering of the sample due to ion bombardment can be neglected as possible inactivation mechanism.	2	1	The other potentially inactivating components of the plasma are not specified in the manuscript. For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description. Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.	1	2	life6020022_perova	1
+The grain size distributions of the three Mars regolith simulants have been added to the text in form of two additional tables (table 2: P-MRS and S-MRS; and table 3: JSC Mars-1A), as well as a new citation for the grain size distribution of JSC Mars-1A [8]. To assess the shading effect of the regolith is difficult. As described we used a very thin layer of regolith particles, but did not measure them according to their shading effect but according to the weight, which was the same for all types of regolith. This experiment was meant as a test of the sterilization method adjusted to our purposes, as described in the introduction section. Therefore, we did not plan it according to the maximal theoretically achievable sterilization effect, but to design a life detection mission to Mars. [8] Wan, L., Wendner, R., & Cusatis, G. A Novel Material for In Situ Construction on Mars: Experiments and Numerical Simulations. 2016, arXiv preprint arXiv:1512.05461	2	1	Three different Mars regolith simulants were used. The mineralogical composition is given in table, but not the grain size distribution. Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.	1	2	life6020022_perova	1
+The title has been changed.	2	1	First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_perova	1
+In the introduction we already described the problem of the destructiveness of heat sterilization like autoclaving on the organic contents of a soil, which was our motivation to test an alternative method such as low temperature plasma sterilization. D. radiodurans was used as test organism because of its resistance towards UV radiation, which is a part of the sterilizing effect of plasma sterilization. The appropriate passages in the introduction section have been edited.	2	1	First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_perova	1
+Since we did not intend to test the influences of the Martian Analog soils on D. radiodurans, but the sterilizing effect of the low temperature plasma treatment on cells intermixed with soil, we did not make a control to determine the effects of the Mars analog soils. The starting values of cell numbers were estimated on samples prepared the same way as the samples for the plasma sterilization (intermixed with soils and desiccated as described in the method section). They were incubated on TYG agar plates like the plasma samples after the treatment. This explanation was missing in the text and has now been added to the appropriate passages in the method section (Chapter 2.2.) for a better understanding.	2	1	The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).	1	2	life6020022_perova	1
+Done, thank you.	2	1	Line 38: Deinococcus radiodurans should be written in italic. While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.	1	2	life6020022_perova	1
+Yes, we just used one medium (TYG), so we changed it to ‘medium’.	2	1	Line 41: Medium instead of media. If I understood correctly, you used only one medium.	1	2	life6020022_perova	1
+The writing has been changed accordingly.	2	1	Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Reply:	1	2	life6020022_perova	1
+Since the bottles containing the Martian Analog soils were autoclaved before use, they should be sterile. We also made a separate test to check for sterility after autoclaving the Martian Analog soils, which has been negative (meaning no contamination could be detected via plating of the autoclaved JSC Mars-1A regolith on TYG medium).	2	1	Line 57: Are you sure that the Martian Analog soils were sterile before using?	1	2	life6020022_perova	1
+An explanation has been added to the text passage.	2	1	Line 78: Please explain in the text what is “a vacuum treatment control”.	1	2	life6020022_perova	1
+The differences might be caused due to the finer grain size of the other regolith used - particularly P-MRS, which possibly increased the shielding effect. This has been added to the discussion section.	2	1	How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?	1	2	life6020022_perova	1
+I added a description at line 38 – 39.	2	1	line 38 - The linking of 'other molecules' to 'phenotype' here is a little odd. There may be a link from one gene to one protein, but one protein is not really a phenotype. There are many quantitative traits affected by many genes.	1	2	life6030029_makarova	1
+I added a description at line 73 – 74.	2	1	line 73 - the point is made that current RNA World theories do not seem compatible with a hydrothermal origin of life. But it is not clear where this leads us. Maybe life did not begin with RNA. Maybe life did not begin with hydrothermal vents. Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable. Maybe there is some slightly different nucleic acid-like polymer that is stable in hydrothermal conditions. Too many unknowns here to be a useful point.	1	2	life6030029_makarova	1
+This comment is in relation to the first comment by reviewer 2. Thus, I added description in lines 99 – 101 and 111 – 112. And I shorten this part to appreciate the comment by reviewer 2.	2	1	lines 93-106 - I mostly agree with this paragraph but it is not said very clearly. Certainly we need to distinguish between long peptides formed spontaneously and those that are translated. But several questions still remain - were there long peptides before long RNA strands? Was there a way of reproducibly making the same amino acid sequence without translating it from RNA? If not, then is there any way that random non-encoded peptides could be useful to RNAs?	1	2	life6030029_makarova	1
+Actually, the comparison was made for cell-based life-like systems. So, I removed 2 references regarding civilization from this part, and add words at line 243 and 276 – 277. And, I simplify descriptions as shown in line 272.	2	1	The idea in caption to Fig 4 that building blocks do not interact directly with the environment seems interesting and plausible, but not very well defined. Are you saying a cell interacts with the environment but a single gene does not? Or a multicellular organism interacts but one cell does not? The citations 52 and 53 are in fields that are not related to biology and origin of life. So I think this idea needs to be explained and justified in a biological context.	1	2	life6030029_makarova	1
+This part is important. While it was difficult to simplify, but I tried to simplify section 2.3 as possible and added some descriptions.	2	1	Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point. The abstract promised to talk about RNA World and the origin of life. At this point the article seems to have strayed a long way from this intention. From the origin of life viewpoint, even the simplest of these (the prokaryote) is still very complex. The issue for the origin of life is how to get to a prokaryote. I don't really think that looking at social insects and human societies will help much in understanding the origin of life. The ideas of CCSI and CMIO seem interesting, but they are discussed with the high-level examples (pages 10-11), not with molecular and cellular examples. Probably a lot of this could be simplified. It is only when we get to paragraph 3.1 that we get to the point of the article.	1	2	life6030029_makarova	1
+It is not essential that the whole circular network to be incorporated in a life-like system. A part of the network is accelerate by a newly evolved ribozyme is important. I tried to describe this, but it seems not sufficient. So I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 48	2	1	Fig 7 - Viewing a metabolism as a cycle seems to be only half the story. Nutrients go in and waste comes out. This is a throughput, not a cycle. Also the diagram does not indicate whether the ribozymes are made by the cycle. I think there is some room for improvement in this diagram.	1	2	life6030029_makarova	1
+I added a description at lines 538 – 540.	2	1	Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.	1	2	life6030029_makarova	1
+As I mentioned above, I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 488.	2	1	Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included. For example there must be ways of making nucleotides from scratch. It cannot be true that the only source of nucleotides is by degrading oligomers.	1	2	life6030029_makarova	1
+Formation of membrane is very important as reviewer 1 mentioned. I added a description at lines 542 – 544, 483 – 488, 544 – 548. line 554 - The case that two genes were essential initially is not made.	2	1	Section 3.4 - 'There is a debate whether CMIO could have formed by a CCSI consisting entirely or mainly of RNA'. This section seems to mix up two important questions. (i) Are RNA catalysts sufficient? Do we need other kinds of biomolecules like protein catalysts? (ii) Do we need the RNA system to be enclosed in a cell membrane or other kind of compartment? If so, can RNA control the growth and division of the membrane? These issues need to be considered separately.	1	2	life6030029_makarova	1
+Polymerase can only amplify the population of RNA molecules. So, the population of RNA molecules would become of a population consisting of most rapid molecules for replication and then further evolution will struggle. Thus, connection of CCSI with CMIO is essential process to escape from such a non-living state.	2	1	line 554 - The case that two genes were essential initially is not made. Can we not have an RNA polymerase that uses other copies of itself as a template? If chemistry supplies monomers, then only one gene is necessary. The input and output would not necessarily need to be controlled by another gene. It would seem difficult to evolve two separate ribozyme functions at the same time.	1	2	life6030029_makarova	1
+As I agree the reviewer 1, this is important point. And it should be evaluate to identify what process was accelerated at the beginning.	2	1	If there were two genes, what were the functions of the two genes? The connection between the reactions catalyzed by the ribozymes and the concepts of CCSI and CMIO is not very clear at this point.	1	2	life6030029_makarova	1
+I remove some descriptions about hydrothermal environments from Abstract and Conclusions.	2	1	The conclusion comes back to the point about primitive Earth environment. This links to the introductory section about hydrothermal conditions etc. I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question. It is only mentioned in the introduction and conclusions.	1	2	life6030029_makarova	1
+This comment is in relation to the thrid comment by reviewer 1. But, reviewer 1 agrees this part. So, I shorten this part.	2	1	One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused. It is also fairly rambling: for example, lines 95-106 could be removed,	1	2	life6030029_makarova	1
+I remove the reference 11 from this part, and moved to line 108. I changed “soft molecules” to “organic molecules”, and changed to “formation and degradation”. The statement was changed to a sentence “… to focus …” shown in line 227.	2	1	"The English is a problem too: ""soft molecules"" is not a typical way of describing organic molecules, and ""formation and deformation"" of molecules would be better expressed as formation and degradation. Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."	1	2	life6030029_makarova	1
+8. This is related to the comments about Fig. I added ref 74, 75. In addition, for the size of ribozyme, I added description at line 565 – 566 and ref 67 – 69.	2	1	More fundamentally perhaps, the distinction that is made between CCSI and CMIO is, as the author himself notes, remarkably similar to that between genotype and phenotype, and information and function, and even replication and metabolism. The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol. 364:249-59). Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.	1	2	life6030029_makarova	1
+I added a description at line 38 – 39.	2	1	line 38 - The linking of 'other molecules' to 'phenotype' here is a little odd. There may be a link from one gene to one protein, but one protein is not really a phenotype. There are many quantitative traits affected by many genes.	1	2	life6030029_perova	1
+I added a description at line 73 – 74.	2	1	line 73 - the point is made that current RNA World theories do not seem compatible with a hydrothermal origin of life. But it is not clear where this leads us. Maybe life did not begin with RNA. Maybe life did not begin with hydrothermal vents. Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable. Maybe there is some slightly different nucleic acid-like polymer that is stable in hydrothermal conditions. Too many unknowns here to be a useful point.	1	2	life6030029_perova	1
+This comment is in relation to the first comment by reviewer 2. Thus, I added description in lines 99 – 101 and 111 – 112. And I shorten this part to appreciate the comment by reviewer 2.	2	1	lines 93-106 - I mostly agree with this paragraph but it is not said very clearly. Certainly we need to distinguish between long peptides formed spontaneously and those that are translated. But several questions still remain - were there long peptides before long RNA strands? Was there a way of reproducibly making the same amino acid sequence without translating it from RNA? If not, then is there any way that random non-encoded peptides could be useful to RNAs?	1	2	life6030029_perova	1
+Actually, the comparison was made for cell-based life-like systems. So, I removed 2 references regarding civilization from this part, and add words at line 243 and 276 – 277. And, I simplify descriptions as shown in line 272.	2	1	The idea in caption to Fig 4 that building blocks do not interact directly with the environment seems interesting and plausible, but not very well defined. Are you saying a cell interacts with the environment but a single gene does not? Or a multicellular organism interacts but one cell does not? The citations 52 and 53 are in fields that are not related to biology and origin of life. So I think this idea needs to be explained and justified in a biological context.	1	2	life6030029_perova	1
+This part is important. While it was difficult to simplify, but I tried to simplify section 2.3 as possible and added some descriptions.	2	1	Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point. The abstract promised to talk about RNA World and the origin of life. At this point the article seems to have strayed a long way from this intention. From the origin of life viewpoint, even the simplest of these (the prokaryote) is still very complex. The issue for the origin of life is how to get to a prokaryote. I don't really think that looking at social insects and human societies will help much in understanding the origin of life. The ideas of CCSI and CMIO seem interesting, but they are discussed with the high-level examples (pages 10-11), not with molecular and cellular examples. Probably a lot of this could be simplified. It is only when we get to paragraph 3.1 that we get to the point of the article.	1	2	life6030029_perova	1
+It is not essential that the whole circular network to be incorporated in a life-like system. A part of the network is accelerate by a newly evolved ribozyme is important. I tried to describe this, but it seems not sufficient. So I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 48	2	1	Fig 7 - Viewing a metabolism as a cycle seems to be only half the story. Nutrients go in and waste comes out. This is a throughput, not a cycle. Also the diagram does not indicate whether the ribozymes are made by the cycle. I think there is some room for improvement in this diagram.	1	2	life6030029_perova	1
+I added a description at lines 538 – 540.	2	1	Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.	1	2	life6030029_perova	1
+As I mentioned above, I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 488.	2	1	Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included. For example there must be ways of making nucleotides from scratch. It cannot be true that the only source of nucleotides is by degrading oligomers.	1	2	life6030029_perova	1
+Formation of membrane is very important as reviewer 1 mentioned. I added a description at lines 542 – 544, 483 – 488, 544 – 548. line 554 - The case that two genes were essential initially is not made.	2	1	Section 3.4 - 'There is a debate whether CMIO could have formed by a CCSI consisting entirely or mainly of RNA'. This section seems to mix up two important questions. (i) Are RNA catalysts sufficient? Do we need other kinds of biomolecules like protein catalysts? (ii) Do we need the RNA system to be enclosed in a cell membrane or other kind of compartment? If so, can RNA control the growth and division of the membrane? These issues need to be considered separately.	1	2	life6030029_perova	1
+Polymerase can only amplify the population of RNA molecules. So, the population of RNA molecules would become of a population consisting of most rapid molecules for replication and then further evolution will struggle. Thus, connection of CCSI with CMIO is essential process to escape from such a non-living state.	2	1	line 554 - The case that two genes were essential initially is not made. Can we not have an RNA polymerase that uses other copies of itself as a template? If chemistry supplies monomers, then only one gene is necessary. The input and output would not necessarily need to be controlled by another gene. It would seem difficult to evolve two separate ribozyme functions at the same time.	1	2	life6030029_perova	1
+As I agree the reviewer 1, this is important point. And it should be evaluate to identify what process was accelerated at the beginning.	2	1	If there were two genes, what were the functions of the two genes? The connection between the reactions catalyzed by the ribozymes and the concepts of CCSI and CMIO is not very clear at this point.	1	2	life6030029_perova	1
+I remove some descriptions about hydrothermal environments from Abstract and Conclusions.	2	1	The conclusion comes back to the point about primitive Earth environment. This links to the introductory section about hydrothermal conditions etc. I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question. It is only mentioned in the introduction and conclusions.	1	2	life6030029_perova	1
+This comment is in relation to the thrid comment by reviewer 1. But, reviewer 1 agrees this part. So, I shorten this part.	2	1	One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused. It is also fairly rambling: for example, lines 95-106 could be removed,	1	2	life6030029_perova	1
+I remove the reference 11 from this part, and moved to line 108. I changed “soft molecules” to “organic molecules”, and changed to “formation and degradation”. The statement was changed to a sentence “… to focus …” shown in line 227.	2	1	"The English is a problem too: ""soft molecules"" is not a typical way of describing organic molecules, and ""formation and deformation"" of molecules would be better expressed as formation and degradation. Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."	1	2	life6030029_perova	1
+8. This is related to the comments about Fig. I added ref 74, 75. In addition, for the size of ribozyme, I added description at line 565 – 566 and ref 67 – 69.	2	1	More fundamentally perhaps, the distinction that is made between CCSI and CMIO is, as the author himself notes, remarkably similar to that between genotype and phenotype, and information and function, and even replication and metabolism. The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol. 364:249-59). Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.	1	2	life6030029_perova	1
+To the best of our knowledge, rare earth doped yttrium iron garnet is still the main material used for magneto-optical devices. They include bismuth doped YIG, bismuth, terbium doped YIG and cerium doped YIG. Other materials such as Wely semimatels are still under theoretical study[1]. We have added comments to these materials in the manuscript. Revisions: Page 1, line 21, added “At present, rare earth doped yttrium iron garnet (RIG) is the most widely used magneto-optical material in integrated MO devices.” Point 2:	2	1	It is not indicated in the introduction whether any alternative to thin films of yttrium iron garnet is currently being considered for use as magneto-optical resonators.	1	2	ma15051691_makarova	1
+Absoultly there is also temperature dependence of the Faraday rotation when shift to other wavelengths. However, the transparency window of this material is in the 1550 nm wavelength range. When moving to shorter wavelengths, the absorption of this material increases sharply, making them less practical for photonic device applications.	2	1	How will the temperature dependence of the Faraday rotation hysteresis loops of Dy:CeYIG thin films change when moving to another wavelength range? Is there any dependence on the wavelength?	1	2	ma15051691_makarova	1
+previously reported that the compensation temperature of DyIG is Tcomp = 225 K [2,3], which could decrease the Faraday rotation angle of Dy:CeIG in this work at the temperature below 300 K. Therefore, a higher Faraday rotation may be related to the increase of the saturation magnetization of this material in this temperature range. Revisions:Page 5, line 170-173, added “The increase of the Faraday rotation below 40 ℃ is possibly due to the increase of the magnetization of this material at this temperature range, considering a compensation temperature of 225 K in Dy3Fe5O12 [19,20].”. References: Okamura, Y.; Minami, S.; Kato, Y.; Fujishiro, Y.; Kaneko, Y.; Ikeda, J.; Muramoto, J.; Kaneko, R.; Ueda, K.; Kocsis, V.; et al. Giant magneto-optical responses in magnetic Weyl semimetal Co3Sn2S2. Nat Commun 2020, 11, 4619, doi:10.1038/s41467-020-18470-0. Sayetat, F. Huge magnetostriction in Tb3Fe5O12, Dy3Fe5O12, Ho3Fe5O12, Er3Fe5O12 garnets. Journal of magnetism and magnetic materials 1986, 58, 334-346.	2	1	How can one explain the increase in the value of the Faraday rotation angle in the range of 30-40 degrees for Dy:CeYIG (Fig. 3f), which is absent for Ce:YIG (Fig. 3f)?	1	2	ma15051691_makarova	1
+We have updated the references and revised this in the context. Revisions: Page 1, line 21, revised to “for silicon integrated photonic circuits (PICs) [1-3].”. Page 1, line 24, revised to “including optical isolators [4-6]”. Page 1, line 41, revised to “which results in reduced bandwidth and isolation ratio [13].”. Page 1, line 45, revised to “in a temperature range of 20-60 ℃ [10].”. Page 5, line 148, revised to “measure the temperature stability of Dy:CeIG and Ce:YIG films[11].”.	2	1	"For several references, the text ""Error! Reference source not found"" appears. Please check with the editor if this can be fixed."	1	2	ma15051691_makarova	1
+We have revised this in the context. Revisions: Page 3, line 98, revised to “where is the Faraday rotation angle of MO films at room temperature. is the NRPS at room temperature.” Point 3:	2	1	In equation (2) and the following line, there are inconcistencies in the use of capital letters for the Faraday rotation angles.	1	2	ma15051691_makarova	1
+Thanks for correcting this. “X-ray diffraction spectra” has been revised into “X-ray diffraction patterns”. Revisions: Page 3, line 109, revised to “X-ray diffraction patterns”.	2	1	"Line 111. ""X-ray diffraction patterns"", not ""spectra""."	1	2	ma15051691_makarova	1
+Thanks for the commerts. The saturation magnetization of rare-earth doped YIG affects the Faraday rotation angle. Sayetat et al. and Ostorero et al. previously reported that the compensation temperature of DyIG is Tcomp = 225 K [1,2], which could decrease the Faraday rotation angle of Dy:CeIG in this work at the temperature below 300 K. Therefore, a higher Faraday rotation may be related to the increase of the saturation magnetization of this material in this temperature range. Revisions: Page 5, line 170-173, added “The increase of the Faraday rotation below 40 ℃ is possibly due to the increase of the magnetization of this material at this temperature range, considering a compensation temperature of 225 K in Dy3Fe5O12 [19,20].”.	2	1	Figures 2 (c) and (d). While the Faraday rotation angle appears to decrease more or less linearly with temperature in the Ce:YIG film (as far as can be inferred from the non-linear horizontal scale), the decrease is not taking place at the same rate in the Dy:Ce:YIG film. What causes this different behavior?	1	2	ma15051691_makarova	1
+Ostorero, J.; Escorne, M.; Pecheron‐Guegan, A.; Soulette, F.; Le Gall, H. Dy3Fe5O12 garnet thin films grown from sputtering of metallic targets. Journal of Applied Physics 1994, 75, 6103-6105. Thanks for pointing this out. We have revised the Figure 3. Revisions: Page 6, revised the horizontal scale of Figure 3 (c) and updated Figure 3 (c). References: Sayetat, F. Huge magnetostriction in Tb3Fe5O12, Dy3Fe5O12, Ho3Fe5O12, Er3Fe5O12 Journal of magnetism and magnetic materials 1986, 58, 334-346.	2	1	Figures 3 (a) and (b): It could be good to use the same horizontal scale for a clearer comparison.	1	2	ma15051691_makarova	1
+Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS) obey the same Bragg's Law (2dsinθ=nλ), but with different distance from sample to detector. With a distance about 384 mm in this work, the WAXS can measure the structure from 0.12 nm to 0.45 nm, which correspond to the crystal lattice, while SAXS can measure the structure from 10 nm to 80 nm, with a distance of 2484 mm. In a SAXS curve, the strength reflect the number of defects, while the q range reflect the size of the defects.	2	1	How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_makarova	1
+Thanks for your question, we looked up several literature and couldn't find a fixed usage of cracks and voids, and in this article, we think that using pores would completely cover our research objectives, so we deleted the cracks in this paper. Computed microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	The authors mention cracks as well as voids. How are the cracks differentiated from the voids? Were voids observed? Could computed microtomography be used to confirm some of the experimental results?	1	2	ma15124258_makarova	1
+Yes, the pores are assumed to be spherical in the fitting of SAXS data, because the spherical model is the simplest and widely used model in the SAXS fitting. The voids are cylindrical that is close to a spherical, so we can get a relatively accurate size distribution by assuming to spherical.	2	1	Are your pores assumed to be spherical? Are cracks cylindrical?	1	2	ma15124258_makarova	1
+Yes, we had an experiment of differential scanning calorimetry (DSC) to characterize the process, as shown in Figure 1. According to the result from DSC, there were an endothermic peak of phase transformation between the temperature range of 151~172 ℃, which are not exactly consistent with the results from in this paper. We think the difference in temperature is result from the different sample amount and heating rate. As with X-ray diffraction (XRD), the WAXS can be used to characterize the content of different phase in the phase transformation. So, we think the results in our paper is reasonable.	2	1	I would be interesting to have additional support for the process by using thermal experiments such as differential scanning calorimetry (DSC). How do you results compare to phase transformation measured using DSC? Are the onsets and finishing of the phase transformations consistent with the DSC measurements?	1	2	ma15124258_makarova	1
+SAXS can be used to measure the defect size and number of defects with rational theory and adequate validation in many	2	1	Do your experiments (WAXS and SAXS) measure the defect size and number of defects or is a model used to infer these quantities?	1	2	ma15124258_makarova	1
+The colors in figure 1 and 3 represent the strength of scattering, while the colors in other figures just represent different date.	2	1	What do the colors in your figures represent?	1	2	ma15124258_makarova	1
+The result of SAXS is a reflection of two-phase with different electron density. In our paper, the CL-20 powders are immersed in GPL107 with a approximately equal electron density with CL-20 crystals, so the SAXS reflects the nano-scale pores inside CL-20 powders. Thereby, the increase of strength in SAXS curve reflects the increase in number of internal nano-scale pores. According to the principle of SAXS, the scale of defects is correlated with the q range, so the growth into larger defects will appear in other q range. X-ray microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	Could there be any other explanation rather than increase in number of internal nano-scale pores? Could the increase be due to larger defects rather than an increase in the number of defects? If so, how do you prove this? Could you use X-ray microtomography on your samples?	1	2	ma15124258_makarova	1
+The GPL107 is resistant to high temperatures about 400 ℃, and we used the SAXS patterns of GPL107 at different temperature as the background to correct any bias at different temperature.	2	1	Is the temperature high enough to cause decomposition effects? If so, do the decomposition products form bubbles in your solvent? If so, could this bias your conclusions?	1	2	ma15124258_makarova	1
+According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity in lager q range can be explained into the increase in more small voids. And also, our fitting result can provide more obvious display.	2	1	Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?	1	2	ma15124258_makarova	1
+In our paper, spherical model was just a choice in the SAXS date fitting, so we did not provide more illustrate, but a reference (L A Feigin and D I Svergun, Structure Analysis by Small-Angle X-Ray and Neutron Scattering, Plenum Press, New York, 1987) was provide in the manuscript. On line 173, we gave a conclusion “the number of pores increased, but the size distribution did not change.” We did not consider the pore volume, but we can deduce that the pores volume increase as the increase of the number of pores.	2	1	"Could you be more specific on the ""spherical model"" on line 170? Co you assume the pore volume remains unchanged, but the number increases? or visa verse?"	1	2	ma15124258_makarova	1
+Thanks again for your attention, According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity without the change of q range can be explained into the number of pores increasing, while the existing pores increasing in volume would result into an change of q to low range.	2	1	Again, what is happening? Are the number of pores increasing or is the existing pores increasing in volume?	1	2	ma15124258_makarova	1
+Thanks for your comments, we added three reference about recent work published in 2021 and 2022.	2	1	I also noticed that your references did not include any of the more recent work published in 2021 or 2022. There are some more recent articles that would be worth citing.	1	2	ma15124258_makarova	1
+Thanks for you comment, I have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety after thermal treatment in the conclusion.	2	1	In the introduction, the authors foreground their work, focusing on the relationship between the presence of pores and sensitivity. It indeed the case, but I could not find any discussion on this issue. Therefore, I suggest include some comments on this and, probably, provide some discussion of how to avoid the increase of such defects, which increase sensitivity of energetic crystals, in particular CL-20.	1	2	ma15124258_makarova	1
+Thanks for you comments, and I have added the particle size in the manuscript.	2	1	Section 2.1. Materials and instruments, you need to specify the particle size of the initial CL-20 powders.	1	2	ma15124258_makarova	1
+The electron density of CL-20 and GPL107 are calculated according to the classical electron radius ( with a size of 2.818 E-13 cm) and the electron number per volume, the detailed information can refer to our previous work (Molecules 2020, 25 , 443.)	2	1	“The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.	1	2	ma15124258_makarova	1
+In our previous work (Molecules 2020, 25, 443. ), we described in detail how to calculate the specific surface area and volume fraction. In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.	2	1	Page 6. 199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.	1	2	ma15124258_makarova	1
+“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20. To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.” We have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after thermal treatment in the conclusion.	2	1	The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.	1	2	ma15124258_makarova	1
+Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS) obey the same Bragg's Law (2dsinθ=nλ), but with different distance from sample to detector. With a distance about 384 mm in this work, the WAXS can measure the structure from 0.12 nm to 0.45 nm, which correspond to the crystal lattice, while SAXS can measure the structure from 10 nm to 80 nm, with a distance of 2484 mm. In a SAXS curve, the strength reflect the number of defects, while the q range reflect the size of the defects.	2	1	How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_perova	1
+Thanks for your question, we looked up several literature and couldn't find a fixed usage of cracks and voids, and in this article, we think that using pores would completely cover our research objectives, so we deleted the cracks in this paper. Computed microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	The authors mention cracks as well as voids. How are the cracks differentiated from the voids? Were voids observed? Could computed microtomography be used to confirm some of the experimental results?	1	2	ma15124258_perova	1
+Yes, the pores are assumed to be spherical in the fitting of SAXS data, because the spherical model is the simplest and widely used model in the SAXS fitting. The voids are cylindrical that is close to a spherical, so we can get a relatively accurate size distribution by assuming to spherical.	2	1	Are your pores assumed to be spherical? Are cracks cylindrical. Are they statistical in nature?	1	2	ma15124258_perova	1
+Yes, we had an experiment of differential scanning calorimetry (DSC) to characterize the process, as shown in Figure 1. According to the result from DSC, there were an endothermic peak of phase transformation between the temperature range of 151~172 ℃, which are not exactly consistent with the results from in this paper. We think the difference in temperature is result from the different sample amount and heating rate. As with X-ray diffraction (XRD), the WAXS can be used to characterize the content of different phase in the phase transformation. So, we think the results in our paper is reasonable.	2	1	I would be interesting to have additional support for the process by using thermal experiments such as differential scanning calorimetry (DSC). How do you results compare to phase transformation measured using DSC? Are the onsets and finishing of the phase transformations consistent with the DSC measurements?	1	2	ma15124258_perova	1
+SAXS can be used to measure the defect size and number of defects with rational theory and adequate validation in many field.	2	1	5. Do your experiments (WAXS and SAXS) measure the defect size and number of defects or is a model used to infer these quantities?	1	2	ma15124258_perova	1
+The colors in figure 1 and 3 represent the strength of scattering, while the colors in other figures just represent different date.	2	1	6. What do the colors in your figures represent?	1	2	ma15124258_perova	1
+The result of SAXS is a reflection of two-phase with different electron density. In our paper, the CL-20 powders are immersed in GPL107 with a approximately equal electron density with CL-20 crystals, so the SAXS reflects the nano-scale pores inside CL-20 powders. Thereby, the increase of strength in SAXS curve reflects the increase in number of internal nano-scale pores. According to the principle of SAXS, the scale of defects is correlated with the q range, so the growth into larger defects will appear in other q range. X-ray microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	7. Could there be any other explanation rather than increase in number of internal nano-scale pores? Could the increase be due to larger defects rather than an increase in the number of defects? If so, how do you prove this? Could you use X-ray microtomography on your samples?	1	2	ma15124258_perova	1
+The GPL107 is resistant to high temperatures about 400 ℃, and we used the SAXS patterns of GPL107 at different temperature as the background to correct any bias at different temperature.	2	1	8. Is the temperature high enough to cause decomposition effects? If so, do the decomposition products form bubbles in your solvent? If so, could this bias your conclusions?	1	2	ma15124258_perova	1
+According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity in lager q range can be explained into the increase in more small voids. And also, our fitting result can provide more obvious display.	2	1	9. Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?	1	2	ma15124258_perova	1
+In our paper, spherical model was just a choice in the SAXS date fitting, so we did not provide more illustrate, but a reference (L A Feigin and D I Svergun, Structure Analysis by Small-Angle X-Ray and Neutron Scattering, Plenum Press, New York, 1987) was provide in the manuscript. On line 173, we gave a conclusion “the number of pores increased, but the size distribution did not change.” We did not consider the pore volume, but we can deduce that the pores volume increase as the increase of the number of pores.	2	1	"10. Could you be more specific on the ""spherical model"" on line 170? Co you assume the pore volume remains unchanged, but the number increases? or visa verse?"	1	2	ma15124258_perova	1
+Thanks again for your attention, According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity without the change of q range can be explained into the number of pores increasing, while the existing pores increasing in volume would result into an change of q to low range.	2	1	12. Again, what is happening? Are the number of pores increasing or is the existing pores increasing in volume?	1	2	ma15124258_perova	1
+Thanks for your comments, we added three reference about recent work published in 2021 and 2022.	2	1	I also noticed that your references did not include any of the more recent work published in 2021 or 2022. There are some more recent articles that would be worth citing.	1	2	ma15124258_perova	1
+Thanks for you comment, I have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety after thermal treatment in the conclusion.	2	1	How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_perova	1
+Thanks for you comments, and I have added the particle size in the manuscript.	2	1	1. Section 2.1. Materials and instruments, you need to specify the particle size of the initial CL-20 powders.	1	2	ma15124258_perova	1
+The electron density of CL-20 and GPL107 are calculated according to the classical electron radius ( with a size of 2.818 E-13 cm) and the electron number per volume, the detailed information can refer to our previous work (Molecules 2020, 25 , 443.)	2	1	2. “The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.	1	2	ma15124258_perova	1
+In our previous work (Molecules 2020, 25, 443. ), we described in detail how to calculate the specific surface area and volume fraction. In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.	2	1	3. Page 6. 199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.	1	2	ma15124258_perova	1
+We have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after thermal treatment in the conclusion. “During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20. To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”	2	1	4. The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.	1	2	ma15124258_perova	1
+There was a question related to electronic devices (eCigarettes) but we missed to add this data. Only 0.5% of participants used eCigaretes or similar devices. We added this sentence in the section Results.	2	1	First is the omission of any items referring to experience/use of eCigarette/vaping. Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.	1	2	medicina58040502_makarova	1
+It has been showed that higher exposure to secondhand smoking increased the risk of smoking. The multivariable logistic regression analysis showed that variables such as secondhand smoke at home, secondhand smoke at faculty, and secondhand smoke at public spaces can determine whether students smoke or not. It is also obvious that non-smokers avoid being in closed smoking places, but this was not subject of this study.	2	1	"Second it a lack of analysis of second-hand smoke exposure. Specifically, whether exposure increases risk of smoking. Correlations do not confirm causality. It is likely that students who smoke actively seek roommates who smoke, and students who do not smoke seek roommates who do not smoke. Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""	1	2	medicina58040502_makarova	1
+Your comment is very logical. Cigarette smoking, sedentary life-style and obesity are the major public health concerns, particularly in the Balkan region. This was a reason that we have provided some additional comments and recommendations for further actions at the very end of the Discussion section.	2	1	Finally, I found it disturbing that medical school faculty tolerate student smoking at all. This is contrary to accepted public health policy internationally. Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous. The link between smoking and reduced life expectancy has been shown to be causal. This result calls into question the university's curriculum addressing the very many diseases associated with tobacco smoking - most of which reduce life expectancy. This is the opposite of health promotion one expects from all healthcare providers - especially physicians.	1	2	medicina58040502_makarova	1
+The “Aim of the study” has been changed completely.	2	1	"The study aim should be more precise ""The aim of this study was...."""	1	2	medicina58040502_makarova	1
+To reply on this comment and to provide readers with more precise data we made some changes in two sections. In the subsection “Sample size” of the section “Materials and methods” we emphasized that “students from 16 faculties of the University of Banja Luka were participants in our study”. In section “Results” we have added a new sentence that “Majority of study participants were medical students (41.4%) and the rest were students from other faculties (58.6%).” Comment 3.	2	1	Study sample - please provide more precise data on population (faculties at the university and its share in the total sample of recruited subjects)	1	2	medicina58040502_makarova	1
+This has been already presented in Table 2. and explained by asterisks (**) below the table: * Students who smoked at least one day in 30 days prior to the survey. ** Students who smoke every day and desired to smoke always or sometimes upon waking up, and where smoking time after waking up was within one day. If necessary the English version of the Questionnaire could be provided as Supplementary material.	2	1	Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status	1	2	medicina58040502_makarova	1
+We have made some changes in the section “Results” to avoid data duplication.	2	1	The results section is too extensive - please avoid describing all the results in the text. Please provide the most important results in the text, and the rest of them are present in Tables.	1	2	medicina58040502_makarova	1
+The number of subjects (N) has been added.	2	1	In table 5 please provide thr number of subjects (n=.....)	1	2	medicina58040502_makarova	1
+The contents of Table 6 and Table 7 have been rearranged and comprised into one table which is now Table 6.	2	1	Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals)	1	2	medicina58040502_makarova	1
+In the last paragraph of the section “Discussion” we have proposed some practical implications and recommendations for further actions.	2	1	Please provide practical implications of this study	1	2	medicina58040502_makarova	1
+The section “Conclusion” has been completely revised and rearranged according to the reviewer’s comment.	2	1	Please provide a conclusion based on obtained findings and please avoid overwhelming conclusions	1	2	medicina58040502_makarova	1
+There was a question related to electronic devices (eCigarettes) but we missed to add this data. Only 0.5% of participants used eCigaretes or similar devices. We added this sentence in the section Results.	2	1	is the omission of any items referring to experience/use of eCigarette/vaping. Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.	1	2	medicina58040502_perova	1
+It has been showed that higher exposure to secondhand smoking increased the risk of smoking. The multivariable logistic regression analysis showed that variables such as secondhand smoke at home, secondhand smoke at faculty, and secondhand smoke at public spaces can determine whether students smoke or not. It is also obvious that non-smokers avoid being in closed smoking places, but this was not subject of this study.	2	1	"lack of analysis of second-hand smoke exposure. Specifically, whether exposure increases risk of smoking. Correlations do not confirm causality. It is likely that students who smoke actively seek roommates who smoke, and students who do not smoke seek roommates who do not smoke. Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""	1	2	medicina58040502_perova	1
+Your comment is very logical. Cigarette smoking, sedentary life-style and obesity are the major public health concerns, particularly in the Balkan region. This was a reason that we have provided some additional comments and recommendations for further actions at the very end of the Discussion section.	2	1	Finally, I found it disturbing that medical school faculty tolerate student smoking at all. This is contrary to accepted public health policy internationally. Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous. The link between smoking and reduced life expectancy has been shown to be causal. This result calls into question the university's curriculum addressing the very many diseases associated with tobacco smoking - most of which reduce life expectancy. This is the opposite of health promotion one expects from all healthcare providers - especially physicians.	1	2	medicina58040502_perova	1
+The “Aim of the study” has been changed completely.	2	1	"The study aim should be more precise ""The aim of this study was...."""	1	2	medicina58040502_perova	1
+To reply on this comment and to provide readers with more precise data we made some changes in two sections. In the subsection “Sample size” of the section “Materials and methods” we emphasized that “students from 16 faculties of the University of Banja Luka were participants in our study”. In section “Results” we have added a new sentence that “Majority of study participants were medical students (41.4%) and the rest were students from other faculties (58.6%).” Comment 3.	2	1	Study sample - please provide more precise data on population (faculties at the university and its share in the total sample of recruited subjects)	1	2	medicina58040502_perova	1
+Response 3. This has been already presented in Table 2. and explained by asterisks (**) below the table: * Students who smoked at least one day in 30 days prior to the survey. ** Students who smoke every day and desired to smoke always or sometimes upon waking up, and where smoking time after waking up was within one day. If necessary the English version of the Questionnaire could be provided as Supplementary material.	2	1	Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status	1	2	medicina58040502_perova	1
+We have made some changes in the section “Results” to avoid data duplication.	2	1	The results section is too extensive - please avoid describing all the results in the text. Please provide the most important results in the text, and the rest of them are present in Tables.	1	2	medicina58040502_perova	1
+The number of subjects (N) has been added.	2	1	In table 5 please provide thr number of subjects (n=.....)	1	2	medicina58040502_perova	1
+The contents of Table 6 and Table 7 have been rearranged and comprised into one table which is now Table 6.	2	1	Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals)	1	2	medicina58040502_perova	1
+In the last paragraph of the section “Discussion” we have proposed some practical implications and recommendations for further actions.	2	1	Please provide practical implications of this study	1	2	medicina58040502_perova	1
+The section “Conclusion” has been completely revised and rearranged according to the reviewer’s comment.	2	1	Please provide a conclusion based on obtained findings and please avoid overwhelming conclusions	1	2	medicina58040502_perova	1
+thank you for reviewer’s comments, we made changes accordingly. We added statistical analysis for CNA association between case and reference groups (please see page 5).	2	1	My main issue is with the way the results are presented. The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value. Even if they performed a test, important questions remain on the selection of the reference. The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry. The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA. My suggestion is to offer a better background for these results by trying to answer two main questions:	1	2	medsci4030012_makarova	1
+For the questions of “is the deletion burden observed really exceptional in a population sample of similar ancestry”, we agree with the reviewer’s comment. We did the analysis a couple of years ago, at that time, there is no CNV data from the 1000 genome for Mexicans, Colombians, Puerto Ricans and Peruvians, however, there is data for the admixed populations, thus we used this for this study. There might be some difficulty to do more analysis since our statistician and bioinformatics person left the department and we are in the process of hiring somebody for this role. At the same, we continue in recruiting more subjects with cervical cancers to further confirm our findings. Moreover, in the discussion, we have list this as one of the limitations in page 8 “2) there might be bias of the CNA identified in cervical cancer for the Mexican Americans since we used the 1000 Genome admixed populations, not Mexican Americans, thus, we current recruit more subjects with cervical cancer from the same population and plan to validate the findings in more samples;” 2)	2	1	1) is the deletion burden observed really exceptional in a population sample of similar ancestry? To answer this question, I would refer to the 1000 genomes data where there are Mexicans, Colombians, Puerto Ricans and Peruvians. There must be available CNV estimations for these ancestries. If you find good estimates then perhaps a statistical tests with those as your control group would be more believable.	1	2	medsci4030012_makarova	1
+For the question of “is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies”, our response is that we did search of CNAs observed in other cancer based on TCGA studies, although there are limited studies, we addressed this issue in the discussion section (page 8) “…Using the TCGA data, a recent study identified nine regions of deletion that were unique to ER+ post menopause tumors in patients with breast cancer, including deletion in 7p22.3 where our newly identified deletion in cases only located and it contains known tumor suppressor gene”.	2	1	2) is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies. Please cite those.	1	2	medsci4030012_makarova	1
+great suggestions, thank the reviewer. Now we added a sentence of “Moreover, the pathway analysis revealed endometrial cancer and estrogen signaling pathways associated with this cancer (P < 0.05) using the KEGG” in the abstract as the reviewer’s suggestion. In addition, in the introduction section, we added “We are aware of the limited number of cases and lack of control group. Thus future a large study with a control sample and more cases as methodological alternative is needed”. (Please see page 3) Minor comments: 1-	2	1	The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract. I suggest bringing this result up to the surface and making it more relevant even from the abstract. You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.	1	2	medsci4030012_makarova	1
+as the reviewer’s suggestion, we changed “Latino” to “Mexican American” in the text of the manuscript and table	2	1	I missed a definition of the sample's ancestry. In the title it refers to Mexican American but in the rest of the manuscript it is treated as a latino population. Note that there are differences in admixture in the latino population depending of their country of origin. Studies on the 1000 genomes show differences between Puerto Ricans, Colombians, Mexicans and Peruvians. Therefore, if the authors can state that their sample is essentially Mexican that would be more informative and precise. I would then limit the use of latino. However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.	1	2	medsci4030012_makarova	1
+now we change “Latino population” to “Mexican American” as the reviewer’s suggestion	2	1	Abstract Line 23: should read Latino Populations	1	2	medsci4030012_makarova	1
+thank you for the comments. We also realized this issue which is due to that the color green overlaps with the other colors, thus it cannot be distinguished, and we have tried different color combinations without help. Now we made changes of the figure and legend.	2	1	Figure 1: I cannot see the supposed deletions marked in green	1	2	medsci4030012_makarova	1
+thanks, we removed the % sign in table cells in Table 2b	2	1	Table 2b: Please remove the % sign in some of the table cells, for instance in Reference (Deletion, 10-100kb).	1	2	medsci4030012_makarova	1
+thank you, reviewer, this is a typo, we corrected it.	2	1	Line 34. The authors need to make clear about “pre-, cancer (91.3%) groups”. It is not clear if there is one or two groups but just one percentage.	1	2	medsci4030012_makarova	1
+yes, we added up to six key words now	2	1	Line 40. The authors may add up to 6 key words.	1	2	medsci4030012_makarova	1
+we made changes based on the reviewer’s suggestion, please see statistical method in page 5 and result in page 6	2	1	Lines 196-197. It is not clear which statistical method is used for the significant results in Table 2.Please give more details.	1	2	medsci4030012_makarova	1
+thanks, we provided definition for pre-cancer in materials, which include patients with CIN I, II, and III.	2	1	Line 199. The authors need to define “pre-cancer” in materials.	1	2	medsci4030012_makarova	1
+thanks, we added “…using statistical analyses described in the method section”	2	1	Please give more details. Lines 214-216. It is not clear which statistical method is used for the significant results.	1	2	medsci4030012_makarova	1
+thanks, the reviewer. Now we removed redundancy statement.	2	1	Lines 238-242 are replicated in discussion lines 376-380. The authors may need to delete the part in lines 238-242.	1	2	medsci4030012_makarova	1
+thank you for reviewer’s comments, we made changes accordingly. We added statistical analysis for CNA association between case and reference groups (please see page 5).	2	1	My main issue is with the way the results are presented. The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value. Even if they performed a test, important questions remain on the selection of the reference. The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry. The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA. My suggestion is to offer a better background for these results by trying to answer two main questions: 1)	1	2	medsci4030012_perova	1
+For the questions of “is the deletion burden observed really exceptional in a population sample of similar ancestry”, we agree with the reviewer’s comment. We did the analysis a couple of years ago, at that time, there is no CNV data from the 1000 genome for Mexicans, Colombians, Puerto Ricans and Peruvians, however, there is data for the admixed populations, thus we used this for this study. There might be some difficulty to do more analysis since our statistician and bioinformatics person left the department and we are in the process of hiring somebody for this role. At the same, we continue in recruiting more subjects with cervical cancers to further confirm our findings. Moreover, in the discussion, we have list this as one of the limitations in page 8 “2) there might be bias of the CNA identified in cervical cancer for the Mexican Americans since we used the 1000 Genome admixed populations, not Mexican Americans, thus, we current recruit more subjects with cervical cancer from the same population and plan to validate the findings in more samples;” 2)	2	1	is the deletion burden observed really exceptional in a population sample of similar ancestry? To answer this question, I would refer to the 1000 genomes data where there are Mexicans, Colombians, Puerto Ricans and Peruvians. There must be available CNV estimations for these ancestries. If you find good estimates then perhaps a statistical tests with those as your control group would be more believable.	1	2	medsci4030012_perova	1
+For the question of “is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies”, our response is that we did search of CNAs observed in other cancer based on TCGA studies, although there are limited studies, we addressed this issue in the discussion section (page 8) “…Using the TCGA data, a recent study identified nine regions of deletion that were unique to ER+ post menopause tumors in patients with breast cancer, including deletion in 7p22.3 where our newly identified deletion in cases only located and it contains known tumor suppressor gene”.	2	1	is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies. Please cite those.	1	2	medsci4030012_perova	1
+Now we added a sentence of “Moreover, the pathway analysis revealed endometrial cancer and estrogen signaling pathways associated with this cancer (P < 0.05) using the KEGG” in the abstract as the reviewer’s suggestion. In addition, in the introduction section, we added “We are aware of the limited number of cases and lack of control group. Thus future a large study with a control sample and more cases as methodological alternative is needed”. (Please see page 3) Minor comments: 1-	2	1	The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract. I suggest bringing this result up to the surface and making it more relevant even from the abstract. You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.	1	2	medsci4030012_perova	1
+as the reviewer’s suggestion, we changed “Latino” to “Mexican American” in the text of the manuscript and table	2	1	I missed a definition of the sample's ancestry. In the title it refers to Mexican American but in the rest of the manuscript it is treated as a latino population. Note that there are differences in admixture in the latino population depending of their country of origin. Studies on the 1000 genomes show differences between Puerto Ricans, Colombians, Mexicans and Peruvians. Therefore, if the authors can state that their sample is essentially Mexican that would be more informative and precise. I would then limit the use of latino. However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.	1	2	medsci4030012_perova	1
+now we change “Latino population” to “Mexican American” as the reviewer’s suggestion	2	1	Abstract Line 23: should read Latino Populations	1	2	medsci4030012_perova	1
+thank you for the comments. We also realized this issue which is due to that the color green overlaps with the other colors, thus it cannot be distinguished, and we have tried different color combinations without help. Now we made changes of the figure and legend.	2	1	Figure 1: I cannot see the supposed deletions marked in green	1	2	medsci4030012_perova	1
+thanks, we removed the % sign in table cells in Table 2b	2	1	Table 2b: Please remove the % sign in some of the table cells, for instance in Reference (Deletion, 10-100kb).	1	2	medsci4030012_perova	1
+thank you, reviewer, this is a typo, we corrected it.	2	1	1) Line 34. The authors need to make clear about “pre-, cancer (91.3%) groups”. It is not clear if there is one or two groups but just one percentage.	1	2	medsci4030012_perova	1
+yes, we added up to six key words now	2	1	2) Line 40. The authors may add up to 6 key words.	1	2	medsci4030012_perova	1
+we made changes based on the reviewer’s suggestion, please see statistical method in page 5 and result in page 6	2	1	3) Lines 196-197. It is not clear which statistical method is used for the significant results in Table 2.Please give more details.	1	2	medsci4030012_perova	1
+thanks, we provided definition for pre-cancer in materials, which include patients with CIN I, II, and III.	2	1	4) Line 199. The authors need to define “pre-cancer” in materials.	1	2	medsci4030012_perova	1
+thanks, we added “…using statistical analyses described in the method section”	2	1	Please give more details. 6) Lines 214-216. It is not clear which statistical method is used for the significant results.	1	2	medsci4030012_perova	1
+thanks, the reviewer. Now we removed redundancy statement.	2	1	7) Lines 238-242 are replicated in discussion lines 376-380. The authors may need to delete the part in lines 238-242.	1	2	medsci4030012_perova	1
+We have attempted to improve the language and hope we have found all significant errors.	2	1	There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.	1	2	molecules26247651_makarova	1
+The characterization of the compounds is provided together with the NMR spectra as Suppl. Mat. S21.	2	1	2. Full characterization of all synthesized compounds is required in the experimental section.	1	2	molecules26247651_makarova	1
+Spectra have been included in Suppl. Mat. S21.	2	1	3. All the 1HNMR and 13CNMR must be attached as supplementary information.	1	2	molecules26247651_makarova	1
+A new Scheme 1 has been produced with the recommended format. The molecular structures in Figures 1 and 3 have been modified accordingly.	2	1	4. Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.	1	2	molecules26247651_makarova	1
+The misalignment is a result of the editorial procedure. We have tried to improve the alignment.	2	1	5. The cell values do not align please correct them.	1	2	molecules26247651_makarova	1
+Done.	2	1	6. Use 4 significant figures for the RNH values in table 2.	1	2	molecules26247651_makarova	1
+We appreciate the suggestion. However, we do not think we have time to perform a proper analysis of XRD data, considering solid state effects, crystal packing, etc.	2	1	7. It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.	1	2	molecules26247651_makarova	1
+In the submitted MSWord document the font was Times New Roman for text and reference numbers. Something has happened during the editorial procedure when changing to Palatino Linotype. Not all reference numbers were changed correctly. We have tried to find and correct the errors.	2	1	8. Check the font of the reference numbers in the text and correct RESPONSE:	1	2	molecules26247651_makarova	1
+This concerns the experimental section, S21, in the Supplementary Material document. We have scrutinized the text and corrected the errors. We have also corrected two typos in the main manuscript.	4	1	Correct subscripts and superscripts in the experimental also check the spacings to be consistent.	3	2	molecules26247651_makarova	1
+The background for the investigation is described in the Introduction section. As indicated by the Reviewer, the present study is an extension of our previous OH···O binding studies. We have for some time wished to investigate the corresponding NH···O systems. The NH···O type of linkage has the advantage that a wide range of compounds can be investigated; in general, systems of the type NH···O=C with an intervening double bond are not tautomeric, in contrast to the corresponding OH···O=C systems. We have added a remark on this aspect to the Introduction section. - The ‘delay’ of 10 years is due to technical and personal factors of no relevance to the reported results.	2	1	In this article, the authors describe the spectral characteristics of NH in the hydrogen bridge NH...O. It is a continuation of similar OH...O binding studies performed 10 years ago. The rationale for undertaking the research is a bit puzzling. I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O. The explanation should be cleared.	1	2	molecules26247651_makarova	1
+"We are aware of the controversial nature of this result. We also expected a linear relationship similar to the one observed by Dudek. However, we have in this work extended the range of compounds to include species of types J, K, and L, apparently resulting in the observed non-linear relationship. The correlation in Fig. 2 involves only experimental values, most of them from the literature, and we have no reason to doubt their reliability. The exponential regression yields R2 = 0.975, SD = 34.3 cm–1, while the corresponding linear regression gives R2 = 0.837, SD = 93.3 cm–1. Hence, the indication of a non-linear correlation is evident. We have added the sentence ""Probably the extension of the range of compounds to include species of the types J, K, and L has led to the non-linear correlation” to the first paragraph on page 13."	2	1	The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.	1	2	molecules26247651_makarova	1
+"Our attention was drawn to Ref. [54] where Grimme et al. state that ""The [DFT-D3] method has been assessed on standard benchmark sets for inter- and intramolecular noncovalent interactions"". We thus found it relevant to try out B3LYP-D3 for the present study of ""intramolecular noncovalent interactions"". But the Reviewer finds that these calculations ""make no sense here"", so we have removed these results from the manuscript. The Reviewer suggests that we check the functional B3PW91 (in the Introduction we actually mention B3PW91 as one of the two functionals recommended by Scott and Radom). We have performed additional B3PW91/6-311++G(d,p) calculations on a series of seven compounds covering a wide range of NH stretching wavenumbers. The computed harmonic wavenumbers are essentially linearly related to those obtained with B3LYP (R2 = 0.9999, SD = 2.6 cm–1). We have added a remark on this result to the last paragraph of Section 3.2."	2	1	Performing calculations with the Grimme dispersion correction did not make sense here, as D3 is important for intermolecular interactions, especially stacking. Here, calculations are performed for isolated molecules. Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.	1	2	molecules26247651_makarova	1
+Corrected!	2	1	183: should be SD = 34.3 cm-1	1	2	molecules26247651_makarova	1
+"The sentence with ""are reasonable"" has been removed."	2	1	"186: ""are reasonable"": Taking into account that (i) this fit is rather unphysical (ii) moving the points by say 50 cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."	1	2	molecules26247651_makarova	1
+The orientation of the axes was chosen according to the trend reported by Novak [56]: Decreasing NH stretching wavenumber corresponds to decreasing isotope ratio.	2	1	Figure 5: Why is the y-axis reversed?	1	2	molecules26247651_makarova	1
+We have attempted to improve the language and hope we have found all significant errors.	2	1	1. There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.	1	2	molecules26247651_perova	1
+The characterization of the compounds is provided together with the NMR spectra as Suppl. Mat. S21.	2	1	2. Full characterization of all synthesized compounds is required in the experimental section.	1	2	molecules26247651_perova	1
+Spectra have been included in Suppl.	2	1	3. All the 1HNMR and 13CNMR must be attached as supplementary information.	1	2	molecules26247651_perova	1
+A new Scheme 1 has been produced with the recommended format. The molecular structures in Figures 1 and 3 have been modified accordingly.	2	1	4. Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.	1	2	molecules26247651_perova	1
+The misalignment is a result of the editorial procedure. We have tried to improve the alignment.	2	1	5. The cell values do not align please correct them.	1	2	molecules26247651_perova	1
+Done.	2	1	6. Use 4 significant figures for the RNH values in table 2.	1	2	molecules26247651_perova	1
+We appreciate the suggestion. However, we do not think we have time to perform a proper analysis of XRD data, considering solid state effects, crystal packing, etc.	2	1	7. It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.	1	2	molecules26247651_perova	1
+In the submitted MSWord document the font was Times New Roman for text and reference numbers. Something has happened during the editorial procedure when changing to Palatino Linotype. Not all reference numbers were changed correctly. We have tried to find and correct the errors.	2	1	8. Check the font of the reference numbers in the text and correct RESPONSE:	1	2	molecules26247651_perova	1
+This concerns the experimental section, S21, in the Supplementary Material document. We have scrutinized the text and corrected the errors. We have also corrected two typos in the main manuscript.	4	1	Correct subscripts and superscripts in the experimental also check the spacings to be consistent.	3	2	molecules26247651_perova	1
+"We are aware of the controversial nature of this result. We also expected a linear relationship similar to the one observed by Dudek. However, we have in this work extended the range of compounds to include species of types J, K, and L, apparently resulting in the observed non-linear relationship. The correlation in Fig. 2 involves only experimental values, most of them from the literature, and we have no reason to doubt their reliability. The exponential regression yields R2 = 0.975, SD = 34.3 cm–1, while the corresponding linear regression gives R2 = 0.837, SD = 93.3 cm–1. Hence, the indication of a non-linear correlation is evident. We have added the sentence ""Probably the extension of the range of compounds to include species of the types J, K, and L has led to the non-linear correlation” to the first paragraph on page 13."	2	1	1) The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.	1	2	molecules26247651_perova	1
+"Our attention was drawn to Ref. [54] where Grimme et al. state that ""The [DFT-D3] method has been assessed on standard benchmark sets for inter- and intramolecular noncovalent interactions"". We thus found it relevant to try out B3LYP-D3 for the present study of ""intramolecular noncovalent interactions"". But the Reviewer finds that these calculations ""make no sense here"", so we have removed these results from the manuscript. The Reviewer suggests that we check the functional B3PW91 (in the Introduction we actually mention B3PW91 as one of the two functionals recommended by Scott and Radom). We have performed additional B3PW91/6-311++G(d,p) calculations on a series of seven compounds covering a wide range of NH stretching wavenumbers. The computed harmonic wavenumbers are essentially linearly related to those obtained with B3LYP (R2 = 0.9999, SD = 2.6 cm–1). We have added a remark on this result to the last paragraph of Section 3.2."	2	1	2) Performing calculations with the Grimme dispersion correction did not make sense here, as D3 is important for intermolecular interactions, especially stacking. Here, calculations are performed for isolated molecules. Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.	1	2	molecules26247651_perova	1
+Corrected!	2	1	183: should be SD = 34.3 cm-1	1	2	molecules26247651_perova	1
+"The sentence with ""are reasonable"" has been removed."	2	1	"186: ""are reasonable"": Taking into account that (i) this fit is rather unphysical (ii) moving the points by say 50 cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."	1	2	molecules26247651_perova	1
+The orientation of the axes was chosen according to the trend reported by Novak [56]: Decreasing NH stretching wavenumber corresponds to decreasing isotope ratio.	2	1	Figure 5: Why is the y-axis reversed?	1	2	molecules26247651_perova	1
+We appreciate the recommendation and comments. We corrected the above-mentioned correction as suggested by the reviewer and presented as follows, 1H NMR (400 MHz, DMSO-d6, ppm): δ 11.43 (s, 2H), 8.34 (m, 2H), 8.28 (m, 1H), 7.65 (d, 2H, J = 4.8 Hz), 7.61 (d, 2H, J = 3.2 Hz), 7.14 (t, 2H, J = 8.8 Hz), 2.51 (s, 6H). 13C NMR (100 MHz, DMSO-d6, ppm): 159.44 (C=O), 154.55 (Ar-C), 148.91 (Ar-C), 143.22 (-C=N-), 140.48 (Ar-C), 130.06 (Ar-C), 129.44 (Ar-C), 128.20 (Ar-C), 125.75 (Ar- C), 15.32 (-CH3).	2	1	The manuscript by Sharmin Akther Rupa et al, the manuscript has many serious concerns on the basis that it cannot be accepted in this journal. for example, 13C NMR must be 100 MHz not 400 MHz, value must be either ascending order or descending order (13C NMR (400 MHz, DMSO-d6, ppm): 1 above example showed the manuscript not written carefully.	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and agree with the reviewer that the computational and experimental results of antimicrobial activity differ to some extent in this part of the study and required further investigation to understand why such activity differs to the ligands by other means in the future. In this study, we successfully synthesized the novel ligands and studied its spectroscopy with other probable studies. Fortunately, we have found outstanding results during the chemo-sensor study and hopefully, we could be able to show the excellent usage of the ligands as a chemo-sensor in near future. Also, we are planning to do a fluorescence imaging study of protein-ligands.	2	1	Second one that is very important for this paper: The activity of L2 molecule against Bacillus Megaterium (PDB 4j6u) was significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which reveals the antimicrobial activity of the molecule. Both compounds are biologically active, but their activity was moderate which did not support their efforts.	1	2	molecules27051656_makarova	1
+We appreciate the comments of the reviewer and the following are the answers in response to the question for the kind consideration of the reviewer- Tyrosinase is a key enzyme in melanogenesis, which is essential for pigmentation. Dysfunction of tyrosinase may cause skin cancer. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-103]. That’s why, to investigate and compare the antimicrobial activity of the synthesized compounds with in vitro data, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium was performed. Also, molecular docking with other proteins of B. Megaterium was conducted, but 46ju provided better binding affinity with the synthesized compounds. For this reason, we aim to study the structure of tyrosinase from the bacteria. Preliminary antimicrobial studies were carried out to validate the docking protocol. At present, we are doing the fluorescence activities of these ligands and L2 showed excellent fluorescent emission at λmax 520 nm. So, we hope that we will carry out ALA scan (Alanine scanning) based MD experiments and 100ns Molecular dynamics (MD) simulation guided protein-ligand stability in the future for our next study since within this shortest time, we are unable to do these experiments. The active binding site of tyrosinase was predicted by CASTp. We have included this in the manuscript as follows: Binding affinity of L1 and L2 The highest anti-bacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in the Fig. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala44A, and ALA44B of tyrosinase enzyme (PDB ID: 4j6u), which also supports the literature [101-103]. Thus, computational results are in good agreement with in vitro antibacterial behaviour of our compounds for novel antibacterial drug design.	2	1	4. The authors did not report any Tyrosinase enzyme inhibition assay, they are requested to justify Tyrosinase inhibition as the probable mechanism of action for these sets of ligands by providing suitable literature references.	1	2	molecules27051656_makarova	1
+We are delighted and thankful for the recommendation of the reviewer.	4	1	In this present study wehave synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and massspectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also doneThis manuscript is suitable publication in Molecules.	3	2	molecules27051656_makarova	1
+We are delighted and thankful for the recommendation.	2	1	The manuscript is well-written and must be accepted in its present form for publication in Molecules.	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and rewrite the abstract as follows, Polytopic organic ligands with hydrazone moiety are in the forefront of new drug research among many others due to their unique and versatile functionality and ease of strategic ligand design. Quantum chemical calculations of these polyfunctional ligands can be carried out in silico to determine the thermodynamic parameters. In this report two new tritopic dihydrazide ligands, N'2, N'6-bis[(1E)-1-(thiophen-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L1) and N'2, N'6-bis[(1E)-1-(1H-pyrrol-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L2) were successfully prepared by the condensation reaction of pyridine-2, 6-dicarboxylic hydrazide with 2-acetylthiophene and 2-acetylpyrrole. The FT-IR, 1H and 13C NMR as well as mass spectra of both L1 and L2 were recorded and analyzed. Quantum chemical calculations were performed at DFT/B3LYP/cc-pvdz/6-311+ G (d, p) level of theory to study the molecular geometry, vibrational frequencies, and thermodynamic properties including changes of ∆H, ∆S, and ∆G for both the ligands. The optimized vibrational frequency and (1H and 13C) NMR obtained by B3LYP/cc-pvdz/6-311 + G (d, p) showed good agreement with experimental FT-IR and NMR data. Frontier molecular orbital (FMO) calculations were also conducted to find the HOMO, LUMO, and HOMO–LUMO gaps of the two synthesized compounds. To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain. In addition, Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium. The outcomes revealed that both L1 and L2 can suppress microbial growth of bacteria and fungi with variable potency. The antibacterial activity results demonstrated the compound L2 to be potentially effective against Bacillus Megaterium with inhibition zones of 12 mm while molecular docking study showed the binding energies for L1 and L2 to be −7.7 and −8.8 kcal mol−1 respectively with tyrosinase from Bacillus megaterium.‪ -	2	1	The conclusion is missing.	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and corrected the font of the references.	2	1	Font of the reference is different from the text	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and corrected the section as below: Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development [1-6]. Because, their polyfunctional nature offer multifarious synthetic ways to derivatize such organic molecules towards suitable and effective drug-receptor interaction. The derivatives of hydrazide-hydrazone moiety specially with heterocyclic system possess a range of biological activities namely, anti-microbial, anti-mycobacterial, antitubercular, anticonvulsant, anticholinesterase [1], antiplatelet, and more importantly antitumor [5,7]. Transition metal complexes derived from such type of ligands have been widely studied since they also demonstrate significant biological and pharmacological properties [8-11].	2	1	This sentence “‪Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development” should be better connected to the next one to explain why they are important for new drug development	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and corrected the section as below: This Section is corrected as “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.’’	2	1	Replace “‪In addition to that, the compounds were tested in vitro bioassays against some Gram-negative and Gram-positive ‪bacteria and the fungus strain showing specially promising results for L2. Molecular docking methodology was used to study molecular behavior of L1 and L2 with Bacillus Megaterium to identify their binding interactions” with “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.‪ Response:	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and added the required references as below. We have these specimens of strains available in our lab to study. Therefore, we used. We also updated the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B and corrected the reference the number of Salmonella Typhi (K-323130) bacteria. We are showing our sincere apology for such mistakes. The changes are added as follows,	2	1	Section 2.3.1. Reference for the used strains is required. Why are those specific strains used?	1	2	molecules27051656_makarova	1
+4.9. We appreciate the recommendation, and corrected this information as shown below: 3.2.	2	1	Please change the info accordingly. ODB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium cristal structure.	1	2	molecules27051656_makarova	1
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation. Protein-ligand Docking 3.2.1. Ligand and Protein preparation: The structures of L1 and L2 have been fully optimized by using Gaussian 09 software at B3LYP/6-311G+ (d, p) level. The 3D crystal structure of tyrosinase from Bacillus Megaterium (PDB ID: 4j6u; resolution: 2.5Å, Chain A, B) was obtained in pdb format from online RCSB protein data bank (PDB) database. The structure was verified, and an energy minimization was performed with the Swiss-Pdb Viewer software packages (version 4.1.0) [44], since the crystal structure contains a variety of issues related to improper bond order, side chains geometry, and missing hydrogen atoms. Prior to docking, all the heteroatoms and water molecules were removed from the crystal structure using PyMol (version 1.3) software packages [45]. The active binding pocket of tyrosinase was predicted by CASTp—having the highest pocket area and volume are 95.432 Å2 and 137.877 Å3, respectively [46]. Both the structures of the proteins and ligands were saved in .pdbqt format by AutoDock Vina (version 1.1.2, May 11, 2011) for docking analysis [47]. 3.2.2. Molecular docking Analysis: The docking calculations were performed using default parameters and 8 docked conformations were generated for both compounds. The energy calculations were done by genetic algorithms. Nonpolar hydrogen atoms, Gasteiger partial charges, rotatable bonds, and grid box with dimensions 66.57 × 58.25 × 84.98 Å3 created on the tyrosinase with the aid of Auto Dock Tools 1.1.2 and spacing of 0.3750 Å. The docked conformation of the respective protein conformer with lowest binding free energy and root mean-square deviation value (RMSD) 0.0 Å was analyzed using PyMOL Molecular Graphics System (version 1.7.4) and Accelrys Discovery Studio 4.1 [49]. We appreciate the recommendation, and splitted the data into 2 more subheads as, 3.2.1. Ligand and Protein preparation and 3.2.2. Molecular docking analysis. Also, more information was added about the software used as shown below: 3.2.	2	1	Section 3.2. and 4.9. The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis. Divide the data into those 2 subheads and add more about the software used to perform the docking and the parameters used (Grid box, extensiveness …) Response:	1	2	molecules27051656_makarova	1
+2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study. We appreciate the recommendation, and this section is updated by changing the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B. Also, the sentences of second part (4.8) are deleted and added to 2.3.1 as suggested by the reviewer. Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method. The formation of diameter of inhibition zones in mm by the synthesized analogues are shown in Table 5. Compound L2 showed moderate activity against Bacillus Megaterium bacteria while L1 showed promising antifungal activity against Aspergillus niger fungal strains compared to standard Amphotericin-B. Diameter of inhibition zones (mm) of the synthesized compounds, Ceftriaxone and Amphotericin-B against tested bacterial and fungal strains.	2	1	Section 4.8. There is no proper description of the results and no discussion in this part. Please revise - This sentence should be moved to the materials and methods or deleted from the section “‪Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (JCM-1652) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) ‪were used in this study.”	1	2	molecules27051656_makarova	1
+Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100]. We appreciate the recommendation and updated the molecular docking study by adding more details and literature references as follows, 4.9.	2	1	The molecular docking study is missing a control to properly analyse the results.	1	2	molecules27051656_makarova	1
+We appreciate the recommendation and changed the conclusion as follows, Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized to present a wide range of biological activities in recent years. In this present study we have synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and mass spectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also done. The computed FT-IR analysis as well as the 1H and 13C NMR using B3LYP/CC-PVDZ/6-311+G(d, p) method agreed satisfactorily with the experimental results. We further evaluated the thermodynamic parameters ∆H, ∆S, and ∆G of the ligands. The geometry optimization revealed the planarity of L1 and L2 molecules. Further, it was seen from the HOMO-LUMO energy values that the chemical potentials were negative and the frontier orbital gap of the molecule under investigation was small, and hence, both compounds are reactive and polarizable. To further showcase the biological activity of the ligands against organic pathogens, the antimicrobial assay was performed and revealed significant inhibition of L2 against Bacillus Megaterium gram positive bacteria, and L1 against Escherichia coli, Aspergillus niger although in lesser extent. The moderate activity of L2 molecule against Bacillus Megaterium is substantiated by molecular docking study against tyrosinase from Bacillus megaterium and was found significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which might suggest the antimicrobial activity of the molecule. Overall, L1 and L2 compounds have spurred significant interest for us from the synthetic, computational and biological point of view. We anticipate continued research regarding these classes of exciting organic ligands.	2	1	Conclusion: Repeated abstract. The conclusion must remind the reader why the article was written in the first place and why it is important in the field. The conclusion should briefly give an insight into the obtained results and also the limitations.	1	2	molecules27051656_makarova	1
+4.9.1. 6. In L1-4j6u, one conventional hydrogen bond (3.04 Å) of O-H----O-C observed between O-H of Tyr267A and O-C group of compounds L1. Pi-cation, pi-sulfur and amide-pi bonds were also noticed with LYS47A, PHE48A, ILE39A, ALA40A, GLY43B and ALA44B, respectively. Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl). L2-4j6u complex was stabilized by four NH….O hydrogen bonds and they were LYS47A (2.25 Å), GLY143B (3.04 Å), Tyr267A (3.07 Å) and PRO219B (2.91 Å) (Fig. 6). Like L1, L2 formed pi-cation and amide-pi bonds with LYS47A, ILE39A, GLY43B, where the distances were 3.73, 4.34, 3.54 Å. L2 also formed seven pi-alkyl bonds with ALA44A (5.19 Å), LYS47A (4.35 Å), ALA44B (3.95 Å), LYS47B (4.98 Å), PRO52A (5.04 Å), ALA40B (4.67 Å) and ILE139B (4.24 Å), respectively. We appreciate the recommendation and rewrote the Molecular Docking study as follows as suggested by the reviewer, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding of ligand-receptor interactions in order to facilitate the design of potential drugs [92-95]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analyses of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [96-99]. The docking results were also compared with well-testified inhibitor arbutin [97]. Binding affinity of L1 and L2 The highest antibacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, whereas for arbutin-4j6u the value was -9.1 kcal mol-1 which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in Fig. It was observed that arbutin formed six conventional hydrogen bonds with 4j6u (Supplementary Fig. S11) by the following residues: ALA40A (one O-H----O-C hydrogen bond), Glu141A (four O-H----O-C hydrogen bond), and LYS47B (one O-H----O-C hydro-gen bond). Also several hydrophobic interactions were found with ILE139A, ILE39A, and ALA40A. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala40A, Ala44A, ALA44B and Lys47B of tyrosinase enzyme (PDB ID: 4j6u), which are in close vicinity to the control arbutin and supports the literature [100-102].	4	1	"The molecular study needs a control molecule to compare the results of L1 and L2. how would we know that −7.7 and −8.8 are good scores or not. Either perform a quick analysis for a molecule known for its tyrosinase inhibition or compare with other studies control that used the same methods in your paper.For exemple in this study ""https://doi.org/10.1016/j.bmc.2011.10.078"" they used Arbutin to compare the results of their studied molecule."	3	2	molecules27051656_makarova	1
+We appreciate the recommendation and corrected the style of the references as suggested by the reviewer.	4	1	The references are in a weird format containing only the author's name, journal name and date. article title and other data are missing. Reference style followed by the journalAuthor 1, A.B. ; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.	3	2	molecules27051656_makarova	1
+We appreciate the recommendation and comments. We corrected the above-mentioned correction as suggested by the reviewer and presented as follows, 1H NMR (400 MHz, DMSO-d6, ppm): δ 11.43 (s, 2H), 8.34 (m, 2H), 8.28 (m, 1H), 7.65 (d, 2H, J = 4.8 Hz), 7.61 (d, 2H, J = 3.2 Hz), 7.14 (t, 2H, J = 8.8 Hz), 2.51 (s, 6H). 13C NMR (100 MHz, DMSO-d6, ppm): 159.44 (C=O), 154.55 (Ar-C), 148.91 (Ar-C), 143.22 (-C=N-), 140.48 (Ar-C), 130.06 (Ar-C), 129.44 (Ar-C), 128.20 (Ar-C), 125.75 (Ar- C), 15.32 (-CH3).	2	1	The manuscript by Sharmin Akther Rupa et al, the manuscript has many serious concerns on the basis that it cannot be accepted in this journal. for example, 13C NMR must be 100 MHz not 400 MHz, value must be either ascending order or descending order (13C NMR (400 MHz, DMSO-d6, ppm): 1 above example showed the manuscript not written carefully.	1	2	molecules27051656_perova	1
+We appreciate the recommendation and agree with the reviewer that the computational and experimental results of antimicrobial activity differ to some extent in this part of the study and required further investigation to understand why such activity differs to the ligands by other means in the future. In this study, we successfully synthesized the novel ligands and studied its spectroscopy with other probable studies. Fortunately, we have found outstanding results during the chemo-sensor study and hopefully, we could be able to show the excellent usage of the ligands as a chemo-sensor in near future. Also, we are planning to do a fluorescence imaging study of protein-ligands.	2	1	Second one that is very important for this paper: The activity of L2 molecule against Bacillus Megaterium (PDB 4j6u) was significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which reveals the antimicrobial activity of the molecule. Both compounds are biologically active, but their activity was moderate which did not support their efforts.	1	2	molecules27051656_perova	1
+We appreciate the comments of the reviewer and the following are the answers in response to the question for the kind consideration of the reviewer- Tyrosinase is a key enzyme in melanogenesis, which is essential for pigmentation. Dysfunction of tyrosinase may cause skin cancer. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-103]. That’s why, to investigate and compare the antimicrobial activity of the synthesized compounds with in vitro data, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium was performed. Also, molecular docking with other proteins of B. Megaterium was conducted, but 46ju provided better binding affinity with the synthesized compounds. For this reason, we aim to study the structure of tyrosinase from the bacteria. Preliminary antimicrobial studies were carried out to validate the docking protocol. At present, we are doing the fluorescence activities of these ligands and L2 showed excellent fluorescent emission at λmax 520 nm. So, we hope that we will carry out ALA scan (Alanine scanning) based MD experiments and 100ns Molecular dynamics (MD) simulation guided protein-ligand stability in the future for our next study since within this shortest time, we are unable to do these experiments. The active binding site of tyrosinase was predicted by CASTp. We have included this in the manuscript as follows: To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100]. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in the Fig. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala44A, and ALA44B of tyrosinase enzyme (PDB ID: 4j6u), which also supports the literature [101-103]. Thus, computational results are in good agreement with in vitro antibacterial behaviour of our compounds for novel antibacterial drug design.	2	1	4. 3. 2. Third one their binding study: 1. Why did the authors consider Tyrosinase from Bacillus megaterium N205A mutant pdb id 4j6u? As they did not report any enzyme specific inhibition related experiments. Docking is a preliminary experiment. How did the author validate the docking protocol? The authors are suggested using some decoy ligands and calculating the enrichment value in order to justify the docking protocol followed by ALA scan (Alanine scanning) based MD experiments. In addition, authors are suggested to report at list 100ns Molecular dynamics (MD) simulation guided protein-ligand stability report followed by either MM-GB or PB-SA based thermo data analysis. The pdb id 4j6u does not contain any bound ligand. How did the author select the binding site? If a few sets of amino acids were considered to define the binding site, then on what basis they select them? The authors did not report any Tyrosinase enzyme inhibition assay, they are requested to justify Tyrosinase inhibition as the probable mechanism of action for these sets of ligands by providing suitable literature references.	1	2	molecules27051656_perova	1
+We are delighted and thankful for the recommendation.	2	1	"Abedin et al. explored ""Synthesis of Novel Tritopic Hydrazone Ligands: Spectroscopy, Biological activity, DFT, and Molecular docking Studies"". The manuscript is well-written and must be accepted in its present form for publication in Molecules."	1	2	molecules27051656_perova	1
+We appreciate the recommendation and rewrite the abstract as follows, Polytopic organic ligands with hydrazone moiety are in the forefront of new drug research among many others due to their unique and versatile functionality and ease of strategic ligand design. Quantum chemical calculations of these polyfunctional ligands can be carried out in silico to determine the thermodynamic parameters. In this report two new tritopic dihydrazide ligands, N'2, N'6-bis[(1E)-1-(thiophen-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L1) and N'2, N'6-bis[(1E)-1-(1H-pyrrol-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L2) were successfully prepared by the condensation reaction of pyridine-2, 6-dicarboxylic hydrazide with 2-acetylthiophene and 2-acetylpyrrole. The FT-IR, 1H and 13C NMR as well as mass spectra of both L1 and L2 were recorded and analyzed. Quantum chemical calculations were performed at DFT/B3LYP/cc-pvdz/6-311+ G (d, p) level of theory to study the molecular geometry, vibrational frequencies, and thermodynamic properties including changes of ∆H, ∆S, and ∆G for both the ligands. The optimized vibrational frequency and (1H and 13C) NMR obtained by B3LYP/cc-pvdz/6-311 + G (d, p) showed good agreement with experimental FT-IR and NMR data. Frontier molecular orbital (FMO) calculations were also conducted to find the HOMO, LUMO, and HOMO–LUMO gaps of the two synthesized compounds. To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain. In addition, Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium. The outcomes revealed that both L1 and L2 can suppress microbial growth of bacteria and fungi with variable potency. The antibacterial activity results demonstrated the compound L2 to be potentially effective against Bacillus Megaterium with inhibition zones of 12 mm while molecular docking study showed the binding energies for L1 and L2 to be −7.7 and −8.8 kcal mol−1 respectively with tyrosinase from Bacillus megaterium.‪ -	2	1	The conclusion is missing.	1	2	molecules27051656_perova	1
+We appreciate the recommendation and corrected the font of the references.	2	1	Font of the reference is different from the text	1	2	molecules27051656_perova	1
+We appreciate the recommendation and corrected the section as below: Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development [1-6]. Because, their polyfunctional nature offer multifarious synthetic ways to derivatize such organic molecules towards suitable and effective drug-receptor interaction. The derivatives of hydrazide-hydrazone moiety specially with heterocyclic system possess a range of biological activities namely, anti-microbial, anti-mycobacterial, antitubercular, anticonvulsant, anticholinesterase [1], antiplatelet, and more importantly antitumor [5,7]. Transition metal complexes derived from such type of ligands have been widely studied since they also demonstrate significant biological and pharmacological properties [8-11].	2	1	This sentence “‪Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development” should be better connected to the next one to explain why they are important for new drug development	1	2	molecules27051656_perova	1
+We appreciate the recommendation and corrected the section as below: This Section is corrected as “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.’’ Section 2.3.1.	2	1	Replace “‪In addition to that, the compounds were tested in vitro bioassays against some Gram-negative and Gram-positive ‪bacteria and the fungus strain showing specially promising results for L2. Molecular docking methodology was used to study molecular behavior of L1 and L2 with Bacillus Megaterium to identify their binding interactions” with “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.‪ Response:	1	2	molecules27051656_perova	1
+We appreciate the recommendation and added the required references as below. We have these specimens of strains available in our lab to study. Therefore, we used. We also updated the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B and corrected the reference the number of Salmonella Typhi (K-323130) bacteria. We are showing our sincere apology for such mistakes. The changes are added as follows, 2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study.	2	1	Reference for the used strains is required. Why are those specific strains used?	1	2	molecules27051656_perova	1
+4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. We appreciate the recommendation, and corrected this information as shown below: 3.2.	2	1	Please change the info accordingly. ODB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium cristal structure.	1	2	molecules27051656_perova	1
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation. Protein-ligand Docking 3.2.1. Ligand and Protein preparation: The structures of L1 and L2 have been fully optimized by using Gaussian 09 software at B3LYP/6-311G+ (d, p) level. The 3D crystal structure of tyrosinase from Bacillus Megaterium (PDB ID: 4j6u; resolution: 2.5Å, Chain A, B) was obtained in pdb format from online RCSB protein data bank (PDB) database. The structure was verified, and an energy minimization was performed with the Swiss-Pdb Viewer software packages (version 4.1.0) [44], since the crystal structure contains a variety of issues related to improper bond order, side chains geometry, and missing hydrogen atoms. Prior to docking, all the heteroatoms and water molecules were removed from the crystal structure using PyMol (version 1.3) software packages [45]. The active binding pocket of tyrosinase was predicted by CASTp—having the highest pocket area and volume are 95.432 Å2 and 137.877 Å3, respectively [46]. Both the structures of the proteins and ligands were saved in .pdbqt format by AutoDock Vina (version 1.1.2, May 11, 2011) for docking analysis [47]. 3.2.2. Molecular docking Analysis: The docking calculations were performed using default parameters and 8 docked conformations were generated for both compounds. The energy calculations were done by genetic algorithms. Nonpolar hydrogen atoms, Gasteiger partial charges, rotatable bonds, and grid box with dimensions 66.57 × 58.25 × 84.98 Å3 created on the tyrosinase with the aid of Auto Dock Tools 1.1.2 and spacing of 0.3750 Å. The docked conformation of the respective protein conformer with lowest binding free energy and root mean-square deviation value (RMSD) 0.0 Å was analyzed using PyMOL Molecular Graphics System (version 1.7.4) and Accelrys Discovery Studio 4.1 [49]. We appreciate the recommendation, and splitted the data into 2 more subheads as, 3.2.1. Ligand and Protein preparation and 3.2.2. Molecular docking analysis. Also, more information was added about the software used as shown below: 3.2.	2	1	The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis. Divide the data into those 2 subheads and add more about the software used to perform the docking and the parameters used (Grid box, extensiveness …) Response:	1	2	molecules27051656_perova	1
+We appreciate the recommendation, and this section is updated by changing the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B. Also, the sentences of second part (4.8) are deleted and added to 2.3.1 as suggested by the reviewer. 2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study. 4.8. Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method. The formation of diameter of inhibition zones in mm by the synthesized analogues are shown in Table 5. Compound L2 showed moderate activity against Bacillus Megaterium bacteria while L1 showed promising antifungal activity against Aspergillus niger fungal strains compared to standard Amphotericin-B. Table 5. Diameter of inhibition zones (mm) of the synthesized compounds, Ceftriaxone and Amphotericin-B against tested bacterial and fungal strains.	2	1	There is no proper description of the results and no discussion in this part.	1	2	molecules27051656_perova	1
+We appreciate the recommendation and updated the molecular docking study by adding more details and literature references as follows, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].	2	1	This sentence should be moved to the materials and methods or deleted from the section “‪Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), ‪two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (JCM-1652) bacteria, ‪and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) ‪were used in this study.”	1	2	molecules27051656_perova	1
+In this present study we have synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and mass spectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also done. The computed FT-IR analysis as well as the 1H and 13C NMR using B3LYP/CC-PVDZ/6-311+G(d, p) method agreed satisfactorily with the experimental results. We further evaluated the thermodynamic parameters ∆H, ∆S, and ∆G of the ligands. The geometry optimization revealed the planarity of L1 and L2 molecules. Further, it was seen from the HOMO-LUMO energy values that the chemical potentials were negative and the frontier orbital gap of the molecule under investigation was small, and hence, both compounds are reactive and polarizable. To further showcase the biological activity of the ligands against organic pathogens, the antimicrobial assay was performed and revealed significant inhibition of L2 against Bacillus Megaterium gram positive bacteria, and L1 against Escherichia coli, Aspergillus niger although in lesser extent. The moderate activity of L2 molecule against Bacillus Megaterium is substantiated by molecular docking study against tyrosinase from Bacillus megaterium and was found significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which might suggest the antimicrobial activity of the molecule. Overall, L1 and L2 compounds have spurred significant interest for us from the synthetic, computational and biological point of view. We anticipate continued research regarding these classes of exciting organic ligands. We appreciate the recommendation and changed the conclusion as follows, 9 Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized to present a wide range of biological activities in recent years.	2	1	The molecular docking study is missing a control to properly analyse the results.	1	2	molecules27051656_perova	1
+4.9.1. Binding affinity of L1 and L2 The highest anti-bacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. 6. In L1-4j6u, one conventional hydrogen bond (3.04 Å) of O-H----O-C observed between O-H of Tyr267A and O-C group of compounds L1. Pi-cation, pi-sulfur and amide-pi bonds were also noticed with LYS47A, PHE48A, ILE39A, ALA40A, GLY43B and ALA44B, respectively. Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl). L2-4j6u complex was stabilized by four NH….O hydrogen bonds and they were LYS47A (2.25 Å), GLY143B (3.04 Å), Tyr267A (3.07 Å) and PRO219B (2.91 Å) (Fig. 6). Like L1, L2 formed pi-cation and amide-pi bonds with LYS47A, ILE39A, GLY43B, where the distances were 3.73, 4.34, 3.54 Å. L2 also formed seven pi-alkyl bonds with ALA44A (5.19 Å), LYS47A (4.35 Å), ALA44B (3.95 Å), LYS47B (4.98 Å), PRO52A (5.04 Å), ALA40B (4.67 Å) and ILE139B (4.24 Å), respectively. We appreciate the recommendation and rewrote the Molecular Docking study as follows as suggested by the reviewer, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding of ligand-receptor interactions in order to facilitate the design of potential drugs [92-95]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analyses of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [96-99]. The docking results were also compared with well-testified inhibitor arbutin [97]. Binding affinity of L1 and L2 The highest antibacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, whereas for arbutin-4j6u the value was -9.1 kcal mol-1 which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in Fig. It was observed that arbutin formed six conventional hydrogen bonds with 4j6u (Supplementary Fig. S11) by the following residues: ALA40A (one O-H----O-C hydrogen bond), Glu141A (four O-H----O-C hydrogen bond), and LYS47B (one O-H----O-C hydro-gen bond). Also several hydrophobic interactions were found with ILE139A, ILE39A, and ALA40A. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala40A, Ala44A, ALA44B and Lys47B of tyrosinase enzyme (PDB ID: 4j6u), which are in close vicinity to the control arbutin and supports the literature [100-102].	4	1	"The molecular study needs a control molecule to compare the results of L1 and L2. how would we know that −7.7 and −8.8 are good scores or not. Either perform a quick analysis for a molecule known for its tyrosinase inhibition or compare with other studies control that used the same methods in your paper.For exemple in this study ""https://doi.org/10.1016/j.bmc.2011.10.078"" they used Arbutin to compare the results of their studied molecule."	3	2	molecules27051656_perova	1
+We appreciate the recommendation and corrected the style of the references as suggested by the reviewer.	4	1	The references are in a weird format containing only the author's name, journal name and date. article title and other data are missing. Reference style followed by the journalAuthor 1, A.B. ; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.	3	2	molecules27051656_perova	1
+We have changed the position of Figure 1 to line 182, and adjusted the arrangement of the figures in the article.	2	1	line. 57-58. It seems that Figure 1 should be placed in other section, namely Section 3.1. , where it is mentioned for the first time (line 176).	1	2	molecules27123843_makarova	1
+We have removed the heading “2. Results” in the revised manuscript.	2	1	"Line 59. ""Remove """"2."" ""Results"""""""	1	2	molecules27123843_makarova	1
+Thank you for your valuable opinion. NAA is an abbreviation for amino acid, and we have added the description of NAA in the section of “2.3 Construction of sensor array”.	2	1	line 148. What does NAA mean?	1	2	molecules27123843_makarova	1
+We have made the appropriate changes in the text to make the sentences more concise and highlighted them in red (line 248).	2	1	"line 251. ""Write """"On TEM images """" instead of """"Transmission electrone microscopy can be observed..."""""""	1	2	molecules27123843_makarova	1
+Thanks for the valuable comment. We have removed the word “that” in the revised manuscript.	2	1	"line 284. ""Remove the word """"that"""""""	1	2	molecules27123843_makarova	1
+We have changed the position of Figure 1 to line 182, and adjusted the arrangement of the figures in the article.	2	1	line. 57-58. It seems that Figure 1 should be placed in other section, namely Section 3.1. , where it is mentioned for the first time (line 176).	1	2	molecules27123843_perova	1
+We have removed the heading “2. Results” in the revised manuscript.	2	1	"Line 59. ""Remove """"2."" ""Results"""""""	1	2	molecules27123843_perova	1
+Thank you for your valuable opinion. NAA is an abbreviation for amino acid, and we have added the description of NAA in the section of “2.3 Construction of sensor array”.	2	1	line 148. What does NAA mean?	1	2	molecules27123843_perova	1
+We have made the appropriate changes in the text to make the sentences more concise and highlighted them in red (line 248).	2	1	"line 251. ""Write """"On TEM images """" instead of """"Transmission electrone microscopy can be observed..."""""""	1	2	molecules27123843_perova	1
+Thanks for the valuable comment. We have removed the word “that” in the revised manuscript.	2	1	"line 284. ""Remove the word """"that"""""""	1	2	molecules27123843_perova	1
+Our studies have shown that the effect of metal vapors on the color of the discharge plasma is much greater in the case of a pulsed breakdown in a nonuniform electric field. This made it possible to compare with the color of various discharges occurring in the upper atmosphere (Figure 1 in the text of the article). Even with well-studied discharges, it is very difficult to obtain such results. Thus, during an arc discharge, although there is a strong evaporation of the electrodes and the ejection of particles of various sizes, the voltage across the gap is low and the discharge in metal vapors outside the high-temperature arc channel is practically absent. Under these conditions, broadband Planck radiation from the arc channel plasma is observed. In a glow discharge, the sputtering of the electrodes has a low rate, and the emission spectra of the plasma of such discharges contain atomic and molecular transitions of the gases used. In the investigated mode, metal vapors are produced and excited by a pulsed discharge.	2	1	Of course, metal vapors, nano- and micro-particles can affect the characteristics of the discharge. This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc. Similar studies and comparisons with known experimental data could be carried out on these objects. In this connection, the use of high-voltage pulse discharges is not justified. The authors draw an analogy with the formation of sprites, then, in my opinion, the study would be more suitable for another journal related, for example, atmospheric research.	1	2	nano12040652_makarova	1
+Plasma concentration discussions have been removed from the text because no such measurements have been made. The nature of the glow of the particles in Figure 3 corresponds to the glow of a micrometeorite that burns down in the Earth's atmosphere [13] and [http://galaxy.astron.kharkov.ua/statti/meteor.htm]. The brightness of the micrometeorite (the particle) glow increases towards the track’s end. This cannot be explained by an increase in the particle velocity, since the particle stops. This occurs after the voltage pulse action. We believe that an increase in the radiation intensity of the particle is due to its heating during deceleration on gas particles. The text of the article has been revised. Such inaccuracies are found throughout the text.	2	1	Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow.	1	2	nano12040652_makarova	1
+The text of the article was finalized and the English was improved.	2	1	In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.	1	2	nano12040652_makarova	1
+The introduction of the manuscript and its text have been revised. In addition, new references have been added to the article. All changes in the text are highlighted in either yellow or red font. First of all, data describing the characteristics of the pulsed nanosecond discharge used were added and the influence of the electrode material was described in more detail. On the other hand, we should note that with a voltage pulse duration from ones to tens of nanoseconds, the experimental conditions differ significantly from the conditions of an electrostatic discharge and the creation of a foggy environment.	2	1	The introduction section could be improved. Some sentences are not well written or convey obvious ideas. Moreover, the aim of this work is to present an overview about the discharge behavior. The introduction would benefit from adding more content and detailed review overview to this research, such as particle lifting in electrostatic discharge , Turbulence effect, Mist-containing environment, Electrode materials.	1	2	nano12040652_makarova	1
+The experimental system was described in more detail, and various modes of discharge ignition were analyzed.	2	1	Please describe better your experimental system, such as ignition energy.	1	2	nano12040652_makarova	1
+In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence. Since under these conditions diffuse discharges, at which the voltage across the gap remains high are formed, metal vapors are excited and ionized together with gas molecules in the discharge gap. This leads to the emission of radiation at various spectral transitions of metal atoms. Only a part of these transitions has a high radiation intensity in the region of interest to researchers. Also, to obtain a high intensity of radiation, transitions of atoms in metal vapors can be used, which are populated as a result of the efficient transfer of energy from excited gas molecules and atoms. Metals, the color of the emission of vapors of which, when excited in the plasma of nanosecond discharges, corresponds to the color of high-altitude atmospheric discharges, were chosen as the material of the electrodes.	2	1	The choice of the electrode material should be clearly explained in the present paper.	1	2	nano12040652_makarova	1
+The color of the glow of the discharge plasma at electrodes made of various metals is associated with excitation certain energy levels of particles in the vapors of these metals. So, when using electrodes made of aluminum, steel and copper, we observed the glow of red, blue and green colors, respectively. The different colors of the discharge when changing the material of the electrodes are determined not by the spark or arc stages, but by bright spots on the electrodes, which are formed due to the explosive emission of electrons [Mesyats, G.A. Ecton mechanism of the vacuum arc cathode spot. IEEE transactions on plasma science, 1995, 23(6), pp. 879-883. (DOI: 10.1109/27.476469)]. These areas in the photographs have a bright white color (see, for example, the photographs in Figures 3, 6, 7). In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material. High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap. However, emission of individual particles is determined by their temperature, it corresponds to the Planck radiation and is broadband. We note once again that in this work, to obtain metal vapors, as well as metal nano- and microparticles, a pulsed nanosecond discharge in a non-uniform electric field was used.	2	1	Please explain the mechanism of different color arcs produced by electrodes of different materials.	1	2	nano12040652_makarova	1
+Several references was added.	2	1	The topic is timely and the results provided are interesting. However, most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only. A spectral analysis is given for one discharge only (discharge in air with a copper electrode): more quantitative results, such as a spectral analysis of the other discharges also, would be helpful for the comparison between the discharges produced in the lab and the ones observed in the atmosphere.	1	2	nano12040652_makarova	1
+The main experiments were carried out with discharges in air, and the air pressure was chosen close to the pressures of high-altitude discharges. Other gases, such as argon, were chosen to better demonstrate the discoloration of the discharge.	2	1	A point that should be clarified is the choice of the gases used for the discharges. One would expect a comparison between discharges that happen in gas mixtures of similar composition. Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?	1	2	nano12040652_makarova	1
+The manuscript has been revised. This judgment was removed from the text of the manuscript.	2	1	Page 5, line 168 The authors state: “An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface.” The sense of this sentence is not completely clear to me. Where does the evaporation of the metal take place? It it took place on the particle surface, it should produce a reduction of its size, rather than an increase.	1	2	nano12040652_makarova	1
+Thanks for the advice. We plan to do this in our next work.	2	1	Page 6, line 218 The authors state: “Particles of metal and its compounds with oxygen and nitrogen with a size of 500 nm and less are nonuniformly distributed on the surface of the slide”. Some data should be provided to show the elemental composition of the particles.	1	2	nano12040652_makarova	1
+Our studies have shown that the effect of metal vapors on the color of the discharge plasma is much greater in the case of a pulsed breakdown in a nonuniform electric field. This made it possible to compare with the color of various discharges occurring in the upper atmosphere (Figure 1 in the text of the article). Even with well-studied discharges, it is very difficult to obtain such results. Thus, during an arc discharge, although there is a strong evaporation of the electrodes and the ejection of particles of various sizes, the voltage across the gap is low and the discharge in metal vapors outside the high-temperature arc channel is practically absent. Under these conditions, broadband Planck radiation from the arc channel plasma is observed. In a glow discharge, the sputtering of the electrodes has a low rate, and the emission spectra of the plasma of such discharges contain atomic and molecular transitions of the gases used. In the investigated mode, metal vapors are produced and excited by a pulsed discharge.	2	1	Of course, metal vapors, nano- and micro-particles can affect the characteristics of the discharge. This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc. Similar studies and comparisons with known experimental data could be carried out on these objects. In this connection, the use of high-voltage pulse discharges is not justified. The authors draw an analogy with the formation of sprites, then, in my opinion, the study would be more suitable for another journal related, for example, atmospheric research.	1	2	nano12040652_perova	1
+Plasma concentration discussions have been removed from the text because no such measurements have been made. The nature of the glow of the particles in Figure 3 corresponds to the glow of a micrometeorite that burns down in the Earth's atmosphere [13] and [http://galaxy.astron.kharkov.ua/statti/meteor.htm]. The brightness of the micrometeorite (the particle) glow increases towards the track’s end. This cannot be explained by an increase in the particle velocity, since the particle stops. This occurs after the voltage pulse action. We believe that an increase in the radiation intensity of the particle is due to its heating during deceleration on gas particles. The text of the article has been revised.	2	1	"The article is more descriptive in nature, there is no explanation of physical laws, but which are misleading readers. For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."" One can agree with the influence of the particle size on the track size, but how can an increase in the particle charge affect the track glow? Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow. Such inaccuracies are found throughout the text."	1	2	nano12040652_perova	1
+The text of the article was finalized and the English was improved.	2	1	In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.	1	2	nano12040652_perova	1
+The introduction of the manuscript and its text have been revised. In addition, new references have been added to the article. All changes in the text are highlighted in either yellow or red font. First of all, data describing the characteristics of the pulsed nanosecond discharge used were added and the influence of the electrode material was described in more detail. On the other hand, we should note that with a voltage pulse duration from ones to tens of nanoseconds, the experimental conditions differ significantly from the conditions of an electrostatic discharge and the creation of a foggy environment.	2	1	The introduction section could be improved. Some sentences are not well written or convey obvious ideas. Moreover, the aim of this work is to present an overview about the discharge behavior. The introduction would benefit from adding more content and detailed review overview to this research, such as particle lifting in electrostatic discharge , Turbulence effect, Mist-containing environment, Electrode materials.	1	2	nano12040652_perova	1
+The experimental system was described in more detail, and various modes of discharge ignition were analyzed.	2	1	Please describe better your experimental system, such as ignition energy.	1	2	nano12040652_perova	1
+In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence. Since under these conditions diffuse discharges, at which the voltage across the gap remains high are formed, metal vapors are excited and ionized together with gas molecules in the discharge gap. This leads to the emission of radiation at various spectral transitions of metal atoms. Only a part of these transitions has a high radiation intensity in the region of interest to researchers. Also, to obtain a high intensity of radiation, transitions of atoms in metal vapors can be used, which are populated as a result of the efficient transfer of energy from excited gas molecules and atoms. Metals, the color of the emission of vapors of which, when excited in the plasma of nanosecond discharges, corresponds to the color of high-altitude atmospheric discharges, were chosen as the material of the electrodes.	2	1	The choice of the electrode material should be clearly explained in the present paper.	1	2	nano12040652_perova	1
+The color of the glow of the discharge plasma at electrodes made of various metals is associated with excitation certain energy levels of particles in the vapors of these metals. So, when using electrodes made of aluminum, steel and copper, we observed the glow of red, blue and green colors, respectively. The different colors of the discharge when changing the material of the electrodes are determined not by the spark or arc stages, but by bright spots on the electrodes, which are formed due to the explosive emission of electrons [Mesyats, G.A. Ecton mechanism of the vacuum arc cathode spot. IEEE transactions on plasma science, 1995, 23(6), pp. 879-883. (DOI: 10.1109/27.476469)]. These areas in the photographs have a bright white color (see, for example, the photographs in Figures 3, 6, 7). In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material. High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap. However, emission of individual particles is determined by their temperature, it corresponds to the Planck radiation and is broadband. We note once again that in this work, to obtain metal vapors, as well as metal nano- and microparticles, a pulsed nanosecond discharge in a non-uniform electric field was used.	2	1	Please explain the mechanism of different color arcs produced by electrodes of different materials.	1	2	nano12040652_perova	1
+Several references was added.	2	1	most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only. A spectral analysis is given for one discharge only (discharge in air with a copper electrode): more quantitative results, such as a spectral analysis of the other discharges also, would be helpful for the comparison between the discharges produced in the lab and the ones observed in the atmosphere.	1	2	nano12040652_perova	1
+The main experiments were carried out with discharges in air, and the air pressure was chosen close to the pressures of high-altitude discharges. Other gases, such as argon, were chosen to better demonstrate the discoloration of the discharge.	2	1	A point that should be clarified is the choice of the gases used for the discharges. One would expect a comparison between discharges that happen in gas mixtures of similar composition. Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?	1	2	nano12040652_perova	1
+The manuscript has been revised. This judgment was removed from the text of the manuscript.	2	1	A couple of requests of clarification about specific parts of the text are also provided in the following. Page 5, line 168 The authors state: “An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface.” The sense of this sentence is not completely clear to me. Where does the evaporation of the metal take place? It it took place on the particle surface, it should produce a reduction of its size, rather than an increase.	1	2	nano12040652_perova	1
+Thanks for the advice. We plan to do this in our next work.	2	1	Page 6, line 218 The authors state: “Particles of metal and its compounds with oxygen and nitrogen with a size of 500 nm and less are nonuniformly distributed on the surface of the slide”. Some data should be provided to show the elemental composition of the particles.	1	2	nano12040652_perova	1
+The second part of this analysis, titled: “Body composition assessment in Mexican children and adolescents. Part 2: Cross-validation of three bio-electrical impedance methods against dual X-ray absorptiometry for whole-body and regional body composition”, has already been accepted by Nutrients #1604578. The two analyses were conducted on the same children’s database and closely complement each other. As the accepted manuscript already includes the phrase ‘Part 2’ we prefer to keep ‘Part 1’ here ABSTRACT 2.	2	1	TITLE The “Part 1” in the title, while perhaps intriguing, calls for some explanation of future directions. This does not seem to be addressed in the article, thus there seems to be no reason to include this phrase in the title.	1	2	nu14051073_makarova	1
+Corrected, we adjusted this paragraph together with the next one to give a clearer explanation of the motivation of this study.	2	1	Also, the difference(s) between FFM and LM definitions are nuanced and complex and should either be defined, or alternate text should be used here. It seems that the authors are simply trying to point out that use of the 4C model is not common and that other methods are normally used. Best to clarify this statement. INTRODUCTION P2 L61: FFM is obtained via the 4C model – it is not a surrogate.	1	2	nu14051073_makarova	1
+We have adjusted accordingly. We hope now we can give a clearer explanation about the motivation of this study.	2	1	P2 L69: The aim stated here does not follow the logic of the preceding statements in the paragraph, i.e., what does comparing BC methods have to do with lack of healthcare or technology? Is the focus then on finding low cost, easy to apply methods? The aim (and intent) of the study seems too simplistic as stated here – merely comparing methods. How would the results be applied to the healthcare situation in Mexico? These topics should be tied together or an alternative motivation needs to be presented.	1	2	nu14051073_makarova	1
+Clarified in lines 164.	2	1	Who was asked to assent? MATERIALS AND METHODS P3 L94: This is awkwardly worded.	1	2	nu14051073_makarova	1
+The data used was total body with head, as recommended for the ISCD when using DXA for body composition instead of for bone densitometry. We clarified and referenced this in the text, lines 204-208.	2	1	P3 L125: DXA: Was the head ROI excluded from the analyses (as per recommendation by the ISCD)? If so, it should be stated. If not, DXA analysis should be redone and all relationships recalculated.	1	2	nu14051073_makarova	1
+This was a typo, we corrected it. Line 260.	2	1	P5 L176: “(n=52?? )” Is there uncertainty of the number of participants or is this merely a typo?	1	2	nu14051073_makarova	1
+We corrected (lines 287-288).	2	1	P5 L207: The means of FM would be compared using t-test, not calculated.	1	2	nu14051073_makarova	1
+Indeed, it was like that. We have changed it to show only the data of the 288 subjects (without the 5 outliers).	2	1	Are the n values incorrect or is the entire cohort presented here? RESULTS Table 1: The n values in the column headers add up to 293 rather than 288.	1	2	nu14051073_makarova	1
+This was because of the inclusion of the 5 outliers in table 1, which has been corrected.	2	1	Table 1; Body Composition Variables: The mean values do not all agree with those in supplementary Table 1.	1	2	nu14051073_makarova	1
+We have	2	1	Table 1; MRI subsample: There is no indication of age, BMI, etc. in this group. That information may be helpful, perhaps also as supplementary data.	1	2	nu14051073_makarova	1
+Added, the analysis was for the total sample and by subgroups by age and sex, line 287 14.	2	1	P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).	1	2	nu14051073_makarova	1
+We have corrected as recommended. Table 4 sent to supplementary material as supplementary table 3.	2	1	P12 L283: Is Table 4 necessary? The title (and aim?) of the paper is all about comparing techniques against the 4C model. One also begins to wonder about affecting type 1 errors due to multiplicity of comparisons (debatable but worth considering). It may be worth considering moving Table 4 to the supplementary file and bringing supplementary Table 1 into the main document. If the focus (see the title) of the article is comparisons with the 4C model, then supplementary Table 1 seems to merit more direct attention.	1	2	nu14051073_makarova	1
+Changed as supplementary figure 3.	2	1	As stated above, it may be worth considering moving these Bland-Altman figures to the supplementary file.	1	2	nu14051073_makarova	1
+Changed, line 19 INTRODUCTION 3.	2	1	It should perhaps be the last paragraph in the Discussion. DISCUSSION P15 L393: This paragraph reads more like a conclusion and seem out of place.	1	2	nu14051073_makarova	1
+We have stated in the conclusion that while individual methods show bias relative to the 4-component reference, the high correlations indicate that all the methods perform well in ranking individual children as having high or low FFM and fat mass. This ranking is itself very valuable in routine clinical care, particularly for longitudinal assessment. We have provided new evidence for the Mexican population that all the methods have utility for this purpose. Furthermore, some of the biases for individual methods can be resolved by the publication of method-specific reference data, whereby all data can be converted to method-specific z-scores. Publishing such reference data is a further aim of our project.	2	1	Much of the Discussion addresses strengths and weaknesses of the different BC methods, as if to provide guidance for clinicians in selecting the best techniques according to their needs and capabilities. None of the methods are ideal – all are flawed (in comparison with a gold standard). Yet a firm conclusion seems to be lacking. Concluding that methods differ is not surprising. One may be better served by discussing how the differences in methods (i.e. over or under estimating FM) may affect health assessments. That is, what impact may underestimating FM, by relying on one particular method, have on child care? How critical is the method selection? Somehow this needs to be tied more firmly to the population and environment being studied.	1	2	nu14051073_makarova	1
+added	2	1	"I am not familiar with the Bland-Altman method, so was unclear when I read the methods section whether the text in lines 204-207 was explaining the Bland-Altman method or describing a separate procedure. I take from the results that those sentences were describing the Bland-Altman procedure, and if so, adding some text along the lines of ""In this procedure..."" would be helpful. As it was, I couldn't understand the description provided, and it didn't seem to match the figures, which were simply labeled as FM, so seemed to be a simple plots of FM using 2 methods on first read."	1	2	nu14051073_makarova	1
+Added to supplementary figure 1 and main figures 1 and 2 3.	2	1	"Relatedly, it would have been helpful to have more explanatory titles for the plots--eg ""Differential correlation between methods across levels of FM"". And, the meaning of the trend line was very counter-intuitive, so an interpretive note under the plots would have been helpful. (E.g., ""A positive trend indicates increasing underestimation of FM at high FM levels; a negative trend indicates increasing overestimation of FM at high FM levels."	1	2	nu14051073_makarova	1
+Specified in limitations lines 993-997. Important to say is that ~89% of Mexican population is Hispanic. Only 6.6% belong to indigenous population and 5.9% to Afro-Mexican ethnic groups.	2	1	There is no description of the race and ethnic composition of the sample. I am not well-versed in the ethnic composition of Mexico, however, would it be helpful to know the degree of representation of, for example, youth who are Black, of Indigenous ancestry, and of European ancestry? Or are there other ethnic or cultural groups that should be represented?	1	2	nu14051073_makarova	1
+Corrected. In the previous version we put data of the whole sample of 293 subjects (including 5 outliers that should not been there). We have corrected the data, and now we only present data on the 288 subjects for both tables.	2	1	I didn't understand why the data differed between Table 1 and Supplementary Table 1. Why did Ns (and means/SDs) differ?	1	2	nu14051073_makarova	1
+Added to table 1.	2	1	Not all abbreviations are listed for the table footnotes (FFM, D2O, DXA, LM, BV, ADP)	1	2	nu14051073_makarova	1
+Thank you for your comment, we have tried to make the abstract less fragmented -Theoretical framework: it is very updated.	2	1	Abstract: it is not conventional. It is very “fragmented”. Try to elaborate it again.	1	2	nu14122489_perova	1
+Thanks for your comments, we have found a more updated reference (reference 18), cited in lines 96-98 to explain more the importance of HRQoL.	2	1	Theoretical framework: it is very updated. To be prudent, try to update some references if you find.	1	2	nu14122489_perova	1
+Line 501-503 We have added that we used a large sample size and that this was powerful enough to detect significant differences within sub-groups. We have also added a further description of our ability to capture results in a snapshot of time. I hope this clarifies this aspect.	2	1	Methods. This paper is based on cross sectional method. Can you justify it in depth? This study includes a size of 1139 adolescents.	1	2	nu14122489_perova	1
+Thank you for your comment, the updated figures and tables are now at the end of the paper.	2	1	Results. In Results, the separations of text and Figures and Tables is strange (when a paper does it, the trend is to include it at the end of all the paper). I recommend you to alternate them (text, Figures and Tables in Results). Nevertheless, results are well elaborated.	1	2	nu14122489_perova	1
+Thank you for your comment, we are glad that you think so	2	1	Conclusion and discussion: They are well elaborated.	1	2	nu14122489_perova	1
+Thank you for your comments about the instruments and the associated domains. We have updated these and aimed to explain them in more detail and with more clarity, sections 2.5, 2.6 and 2.7 lines 193-196, 215-225, 236-244.	2	1	The level of reporting of the core assessment instruments and their items, dimensions and subdomains needs to be enhanced.	1	2	nu14122489_perova	1
+These issues have been addressed. The tables and figures are more clear and more detailed. We have provided are reasoning for the choices of statistical methods.	2	1	There is a need to revising the reporting of the results and information in the tables along with the statistical methodology used with interval data and group comparison research.	1	2	nu14122489_perova	1
+Thank you for this concern. We have aimed to explain that the KS-10 takes items from the longer KS versions but does not measure for each dimension as it is a general score. However, you have made a great point that some associations may be lost on the specific dimensions, we have tried to explain this reasoning such the burden of participation is lower to complete the KS-10 version lines 493-494 . We wanted to use the general score so that results can be compared with other studies and in other countries, which is one of the advantages of KS-10. Thank you for the insight, this has made us reflect on the KS-10 from the longer versions more.	2	1	The focus on only the global mean score for the wellness KQ-10 measure rather than the 5 dimensions. Diet is only expected to influence 2 of these 5 dimensions.	1	2	nu14122489_perova	1
+We have now recognised the impact and importance of income to a greater depth and are thankful for your comments on this matter. It is a shame that we cannot say anything about income in this article as this was not measured, only parental education, however this has been suggested for future studies lines 535-537 .	2	1	The lack of recognition that income and cost of living rather than education of parents is also a likely reason for the findings.	1	2	nu14122489_perova	1
+We have now made a recommendation to investigate this in more depth and to compare diet quality between school time and home time. It is important to assess if the lunches are the same across students of different SES, some insight on this is given in lines 522 and 534 and that an issue could be the diet quality provided at home as school lunches may even-out diet-related inequalities. Thank you for the recommendation as this is an important aspect of adolescents’ diet.	2	1	The lack of recommendations associated with the Swedish free lunch program to assist the diets of students in low SES families to have more fruit and vegetables	1	2	nu14122489_perova	1
+Thank you for your comments on this matter, we have edited this and hope it makes more sense now and that it does not make that assumption any longer, see lines 48-50.	2	1	The first sentence assumes that poor diet is always associated with adolescents, this is incorrect Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing.	1	2	nu14122489_perova	1
+We understand your concern, there are a lot of abbreviations, the abbreviations have been re-written in each section to help follow the flow, and we have now added an abbreviation and key word table before the introduction to provide additional help for readers, thank you for this comment lines 37-45.	2	1	Line 90 need to put in the full name Health Related Quality of Life (HRQoL). You have so many abbreviations confusing to follow in places. The authors may know what these are, but many reader will not. To assist the reader’s comprehension, explain the instruments more as you develop the paper.	1	2	nu14122489_perova	1
+Thank you for your great comment, we want the dimensions to be clear for the readers and this is an important observation. We have now edited how the SHEIA and RADDS variables are explained as well as the KS-10, sections 2.5 and 2.6 and we hope they are clearer to understand lines 193-196, 215-225, 236-244.	2	1	The important issue is in the method section the reader needs understand the dimensions and sub-dimension that make up within each survey. Unless these are included the reader can not fully understand the study.	1	2	nu14122489_perova	1
+Thank you for your comments, it is important to mention status and the term SES-F has now been incorporated. We have also now used the term parental education and in section 2.7 we have mentioned that this was used as a proxy for SES, lines 257-258. This is to make it clearer that we only had access to parental education and no other SES-Fs and we hope that this is clearer now, we have used parental education instead of SES in lines 21, 111-114 and 149-150. We have also emphasised the importance of parental income, lines 419-428, 503-506, 533-537 and 550-555.	2	1	The term is usually SES social economic status so it should be SES- F if it is social economic status – factors. Given you are using education of parents as the status measure of SES the term status is important in this paper and should not be dropped.	1	2	nu14122489_perova	1
+Thank you, we agree that income as well as education plays a role in healthy food choices. However, the results in the article we are referring to looked at parental education, not income. As we did not have access to data in parental income, we were not able to include that variable in our paper	2	1	It is more that education it is also income re line 106-107 “It is proposed that parents with more educational experience are more likely to make healthier food choices for their family” . Miss the point It also assumes that parents with more educational experience are more likely to have higher incomes and so are more likely to make and avoid healthier food choices for their family	1	2	nu14122489_perova	1
+Although a higher education might lead to a higher income we unfortunately do not have the data to look at that but as previously mentioned we have now stressed the importance of parental income, lines 419-428, 503-506, 533-537 and 550-555, thank you for pointing this out.	2	1	It also assumes that parents with more educational experience are more likely to have higher incomes and so are more likely to make and avoid healthier food choices for their family.	1	2	nu14122489_perova	1
+Thank you for your comment, income is most certainly important and we have now emphasised its importance in lines 419-428, 503-506, 533-537 and 550-555. We also have mentioned that parental education is being used as a proxy for SES lines 257-258 but we cannot infer anything about income as this was not measured, only parental education, not overall SES.	2	1	If income is not important than the paper should be the on home education level and home diet not home SES and home diet.	1	2	nu14122489_perova	1
+Thank you for suggesting that we reference these two papers, Darmon and Drewnoski was very insightful and most certainly useful for this article, and we now make it clearer that education is a factor of SES and a proxy for income. Lines 419-428 offer an insight into income and affluence, however we have elaborated more on income in future perspectives 533-537 and in the conclusion, lines 550-555 as it may, as you say, play a significant role in these associations.	2	1	See Murayama, N. (2015). Effects of socioeconomic status on nutrition in Asia and future nutrition policy studies. Journal of nutritional science and vitaminology, 61(Supplement), S66-S68. Darmon, N., & Drewnowski, A. (2008). Does social class predict diet quality?. The American journal of clinical nutrition, 87(5), 1107-1117. The issues raised in the well quoted Darmon and Drewnowski paper need to be considered more.	1	2	nu14122489_perova	1
+Thank you for your comment, we want to make the definition of this index as clear as possible and we have edited this in section 2.5.1 and included the sub-components, lines 193-196. We have also cited that reference in both sections 2.5.1 and 2.5.2 so that readers can find a more detailed description. We have also edited section 2.5.2, lines 215-225 so that the RADDS index is also easier to understand and have mentioned some of the sub-components.	2	1	The Swedish Healthy Eating Index for Adolescents 2015 (SHEIA15) is not well explained in this paper. The following paper did a better job of reporting it Moraeus L, Lindroos AK, Warensjö Lemming E, Mattisson I. Diet diversity score and healthy eating index in relation to diet quality and socio-demographic factors: results from a cross-sectional national dietary survey of Swedish adolescents. Public Health Nutr. 2020 Jul;23(10):1754-1765. doi: See their Table 5.	1	2	nu14122489_perova	1
+Thanks for your observation, we have now tried to describe the KS-10 with more details, and to make it clearer, section 2.6, 236-244.	2	1	The KIDSCREEN-10 is not well described. The basic information is not there in the paper: “KIDSCREEN-10 (KS-10) is derived from the KIDSCREEN-27, and provides a single index of global QoL using ten items related to physical well-being, psychological well-being, autonomy and parent relation, social support and peers, and school environment” See how these researchers have described the KIDSCREEN-10 . Bouwmans, C., van der Kolk, A., Oppe, M., Schawo, S., Stolk, E., van Agthoven, M., ... & van Roijen, L. (2014). Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD. The European Journal of Health Economics, 15(9), 967-977.	1	2	nu14122489_perova	1
+We are very pleased that you found this interesting!	2	1	In terms of results the flowchart was interesting.	1	2	nu14122489_perova	1
+Thank you for this comment, the figure may have been minimised and red meat and poultry are now visible which may not have been before, apologies for this, also we have mentioned some of the sub-components in lines 215-225. Also the figures and tables became distorted when the manuscript was uploaded, we have fixed this.	2	1	I am a not European reviewer and so found the RADDS a rather restricted list without some meat.	1	2	nu14122489_perova	1
+Thank you for your comment. However, we think chi-squared is the appropriate statistical test as we are only comparing the proportions of distribution between categorical variables, we are not comparing the means between more than two groups which is what ANOVA is used for. We have not used chi square to assess analysis of variance, we have now made it clearer that frequency distribution is being assessed in the statistical methods section In terms of gender and education: the mean, standard deviation, df and sig t or F test need to be reported in the tables.	2	1	Why was the Pearson's chi-squared test used (table 1) for analysis of variance? For while gender and in this study education are categorical (group) the data being evaluated is continuous and interval data and so an ANOVA or MANOVA by group is the method of analysis of variance. (Tabachnick, B.G., Fidell, L. S., & Ullman, J.B. (2007). Using multivariate statistics (5th ed.). Pearson.)	1	2	nu14122489_perova	1
+Thanks for this comment, we have incorporated t and df scores, see tables 1 and 4.	2	1	In terms of gender and education: the mean, standard deviation, df and sig t or F test need to be reported in the tables.	1	2	nu14122489_perova	1
+This has been put in the appendix, table A1, page 18, thanks for the suggestion The regression analyses “p” value is reported, but the beta values and significance must also be reported.	2	1	Similarly, a correlation matrix is typically reported as it the foundation of regression analyses and so it needs to be reported to understanding the interaction between three main tests variables being investigated in this study.	1	2	nu14122489_perova	1
+Great that you mentioned the beta values, the coefficients in the tables are in fact unstandardised beta coefficient values, this has been made clearer in the tables. The tables have diet (ind variable) on the left and the dependent variable KS-10 is on the top to indicate that interaction, then this is stratified by gender. I hope this makes more sense now.	2	1	The regression analyses “p” value is reported, but the beta values and significance must also be reported. The regressing table needs reworked, as it is the influence of diet the independent variable on wellbeing KO-10, the dependent variable. If the focus is on gender typically both a boy and a girl regression model is reported.	1	2	nu14122489_perova	1
+We had a problem with uploading the figures and they became distorted, thank you for your comments, we have fixed this and incorporated t and df values.	2	1	Table 5 is interesting but the setting out is poor and so the columns do not align with headings, particularly the wellness KQ -10 information. Again, an ANOVA “ t” value and df as well as the p value have to be reported. In table 5 only one p value is reported, but what it is measuring is unclear, as there are a number of interactions occurring. Should be reporting total, then girls, and then boys as there look to be interaction effects.	1	2	nu14122489_perova	1
+We appreciate your concern regarding the KS-10 item domains. The reason we chose to use the general item value is that it is less burdensome than the longer versions and it is best practice to use the general score and not to directly try to analyse for each dimension. We are not sure that it is a fact that diet can not influence parent relations or social relations, if diet can help to improve quality of life then this may improve how an adolescent feels and their emotions which ultimately may have an influence on relations to others. However, we realise that we may miss out other findings and have mentioned this as a limitation, lines 496-497 and we appreciate your views on this matter. We have also incorporated more information about KS-10 reliability in measuring HRQoL, 236-244This is also a cross sectional study and so only associations can be established not causations.	2	1	Because the KQ-10 is a composite tests there is the likelihood to be some interaction with the sub-domains. Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet. Diet is not expected to have any influence of parent relations, social relations or peers but your study may find an influence on psychological well-being and even school environment. This is the core of your study: does diet have an influence on psychological wellbeing? Remember the KQ-10 is made up of five subdomains (psychological well-being, autonomy and parent relation, social support and peers, and school environment) The fact is diet is no expected to change 3 of these KQ-3 subdomains and only one or two KQ-10 dimensions. Use all five of the KO-10 dimensions as your outcome measure not the Mean average global KQ-10 score. You may have a more important study if you do that, with a different finding to what you have just using the composite total.	1	2	nu14122489_perova	1
+Great comments, we have elaborated more on income, lines 419-428, 503-506, 533-537 and 550-555. We have also included a detailed section about the significance of school lunches and education in Sweden across different SES groups lines 528-532. In Sweden the school lunch is of rather high diet quality, reaching many of the national dietary recommendations and is provided free of charge regardless of income or SES. However, it would be insightful to a complete a deeper analysis investigating the differences of school quality across differing socio-demographic areas. Nonetheless, the problem related to diet quality may lie in the food provided at home as financial constraints are most likely to be of more significance, lines 528-534.	2	1	The conclusion is sound given the findings but the lack of reference to income is an issue as educational status of parents is often a “de-facto” measure for income. It maybe, it is the cost of living associated with fresh fruit and vegetables that is the real issue. Given the findings the researchers could be arguing that a review may be needed with the Swedish school lunch program and increase the level of fruit and vegetables in the students' diet, particularly for students in lower SES locations. Schools may be making the lunches to a budget rather than to a healthy diet criteria which is more expensive. Different sub-populations even in the same school may need different mix of foods. A public education program could also be encouraged about health eating.	1	2	nu14122489_perova	1
+No comments to address (‘Yes’ ticked for all assessment criteria). We thank Reviewer 1 for their encouraging feedback.	2	1	The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach. This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI. This study deals with question that needs important discussion for obesity-related public health policy development.	1	2	obesities2010010_makarova	1
+Thank you for your encouraging feedback on the potential reach of the findings. We have added at statement to this effect in the Discussion: Although based on a restricted area of exploration, Australian print news media only, our findings may have much broader significance for worldwide social trends and prompt the need for ongoing analysis of media reporting of obesity and weight-related public health policy. Future research could also extend our word embedding analysis to policy texts themselves, to draw direct correlations between media and policy data sources.	2	1	This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.	1	2	obesities2010010_makarova	1
+We agree that analysis of associations between weight-biased language in news media and the prevalence of eating disorders and other mental disorders correlated with weight stigma would be informative to the weight stigma literature, however our input data for the natural language processing (NLP) were textual data only and do not take into account other data modalities such as images and/or clinical data (floating numbers). In other words, NLP is a text-analytical tool to understand the nuances of human language about a certain topic (obesity in our case). This is done by capturing the contextual relationships between words and sentences in text corpus. Furthermore, this suggestion goes beyond the scope of the present paper, which focuses on associations between language biases tied to individual and structural dimensions of obesity and changes in public health policy rather than associations between language biases and changes in mental disorder prevalence. An alternative approach with the aid of NLP, would be to add mental disorders as a dimension in the analyses, but this would require a comprehensive literature review to make sure the mental disorder keywords, and their dichotomous mappings, were inclusive. Given the turnaround time for the revision (10 days), unfortunately we cannot extend the analysis in this way, but we absolutely agree with Reviewer 2 that this is an important and interesting direction for future research that can be achieved with further application of the techniques we have developed for this paper. We have acknowledged investigation of the association between news reporting on obesity and mental disorders as a fruitful direction for future research in the Discussion as follows: It is also important to examine relationships between news media reporting of obesity and health outcomes over time given, for example, medium to large meta-analytic associations between weight stigma and mental health disorders such as anxiety, depression, eating disorders and other psychopathological symptoms [new citation to be added and numbered accordingly – see below]. Emmer, C., Bosnjak, M., Mata, J. The association between weight stigma and mental health: A meta-analysis. Obesity Reviews 2020, 21:e12935. doi:10.1111/obr.12935 Comment 3:	2	1	I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance. Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).	1	2	obesities2010010_makarova	1
+We have provided new versions of the figure as a separate file to support editing/reproduction.	2	1	Legend for Figure 2 is missing.	1	2	obesities2010010_makarova	1
+The research questions have been moved to the Introduction.	2	1	Please move the research questions from the methods to the introduction.	1	2	obesities2010010_makarova	1
+This text was intended as an overview of what follows in the Results. We have moved it to the beginning of the Results and rephrased it as follows: In this section, we show the associations between obesity-related terms and the gender, healthiness, social status, and stereotype dimensions. These associations are subsequently cross-matched with the obesity policy timeline in the Discussion, to help interpret the context of change in biases over time.	2	1	Last paragraph of the methods: why did you report here your conclusions? Please move this part from this section.	1	2	obesities2010010_makarova	1
+Figure 1 has been removed, along with the following associated text: Data extraction and analysis processes are illustrated in Figure 1.	2	1	Please revised Figure 1 because it's very difficult to read.	1	2	obesities2010010_makarova	1
+We have added the following text from this paper in the first paragraph of the discussion as it seems to fit best with our results: Such entrenched weight biases, persistent in the media, may lead to internalised or self-stigma among individuals with overweight and obesity that persist even after weight loss. A recent study [insert numbered citation] performed a semantic evaluation of body shapes in obesity surgery patients and overweight/obesity controls and found that both groups were more willing to accept positive adjectives as a match when BMI was low and negative adjectives as a match when BMI was high.	2	1	I think this aspect is interesting and linked to your results, showing a connection with clinical data. A recent paper has pointed out that postbariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z).	1	2	obesities2010010_makarova	1
+We have now added a Limitations and Future Research section to the Discussion to address this point and others: There are two limitations in our data curation process, the automated approach we used to check and select papers. Firstly, automatic classifiers of any sort can include some irrelevant or false positive articles. Due to the large amount of articles in our dataset, in Step 2 of our methodology, we developed a machine learning binary classifier – a support vector machine - with 87.56% accuracy to automatically identify relevant articles (accuracy is the number of correct predictions made, divided by the total number of predictions made, and then multiplied by 100 to convert it into a percentage). As a rule of thumb, accuracy of a predictive model that is above 80% is very commonly used to summarise the performance of that model. Still, it doesn’t exclude the possibility of irrelevant articles in our final dataset, but that possibility is less than 12.44 percent and this has to be considered against the benefit of scale and efficiency that this method allows. We also acknowledge that our analysis limited is in not being able to consider visual content visual content, images within articles (known as image framing), which have been shown to carry stigmatising elements [48]. This is something that could be added to the approach by including image classification along with additional measures. Furthermore, even though the Dow Jones is one the largest news databases, it might still miss some articles or news sources (although this doesn’t relate to the automatic approach as such). This applies to social media, even though this would not span as long a timeframe.	2	1	You report that an automatic approach was used to check all the papers. Is there any possible limit with this approach in the selection of the papers?	1	2	obesities2010010_makarova	1
+No comments to address (‘Yes’ ticked for all assessment criteria). We thank Reviewer 1 for their encouraging feedback.	2	1	The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach. This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI. This study deals with question that needs important discussion for obesity-related public health policy development.	1	2	obesities2010010_perova	1
+Thank you for your encouraging feedback on the potential reach of the findings. We have added at statement to this effect in the Discussion: Although based on a restricted area of exploration, Australian print news media only, our findings may have much broader significance for worldwide social trends and prompt the need for ongoing analysis of media reporting of obesity and weight-related public health policy. Future research could also extend our word embedding analysis to policy texts themselves, to draw direct correlations between media and policy data sources.	2	1	This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.	1	2	obesities2010010_perova	1
+We agree that analysis of associations between weight-biased language in news media and the prevalence of eating disorders and other mental disorders correlated with weight stigma would be informative to the weight stigma literature, however our input data for the natural language processing (NLP) were textual data only and do not take into account other data modalities such as images and/or clinical data (floating numbers). In other words, NLP is a text-analytical tool to understand the nuances of human language about a certain topic (obesity in our case). This is done by capturing the contextual relationships between words and sentences in text corpus. Furthermore, this suggestion goes beyond the scope of the present paper, which focuses on associations between language biases tied to individual and structural dimensions of obesity and changes in public health policy rather than associations between language biases and changes in mental disorder prevalence. An alternative approach with the aid of NLP, would be to add mental disorders as a dimension in the analyses, but this would require a comprehensive literature review to make sure the mental disorder keywords, and their dichotomous mappings, were inclusive. Given the turnaround time for the revision (10 days), unfortunately we cannot extend the analysis in this way, but we absolutely agree with Reviewer 2 that this is an important and interesting direction for future research that can be achieved with further application of the techniques we have developed for this paper. We have acknowledged investigation of the association between news reporting on obesity and mental disorders as a fruitful direction for future research in the Discussion as follows: It is also important to examine relationships between news media reporting of obesity and health outcomes over time given, for example, medium to large meta-analytic associations between weight stigma and mental health disorders such as anxiety, depression, eating disorders and other psychopathological symptoms [new citation to be added and numbered accordingly – see below]. Emmer, C., Bosnjak, M., Mata, J. The association between weight stigma and mental health: A meta-analysis. Obesity Reviews 2020, 21:e12935. doi:10.1111/obr.12935 Comment 3:	2	1	I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance. Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).	1	2	obesities2010010_perova	1
+We have provided new versions of the figure as a separate file to support editing/reproduction.	2	1	Legend for Figure 2 is missing.	1	2	obesities2010010_perova	1
+The research questions have been moved to the Introduction.	2	1	Please move the research questions from the methods to the introduction.	1	2	obesities2010010_perova	1
+This text was intended as an overview of what follows in the Results. We have moved it to the beginning of the Results and rephrased it as follows: In this section, we show the associations between obesity-related terms and the gender, healthiness, social status, and stereotype dimensions. These associations are subsequently cross-matched with the obesity policy timeline in the Discussion, to help interpret the context of change in biases over time.	2	1	Last paragraph of the methods: why did you report here your conclusions? Please move this part from this section.	1	2	obesities2010010_perova	1
+Figure 1 has been removed, along with the following associated text: Data extraction and analysis processes are illustrated in Figure 1.	2	1	Please revise Figure 1 because it's very difficult to read.	1	2	obesities2010010_perova	1
+We have added the following text from this paper in the first paragraph of the discussion as it seems to fit best with our results: Such entrenched weight biases, persistent in the media, may lead to internalised or self-stigma among individuals with overweight and obesity that persist even after weight loss. A recent study [insert numbered citation] performed a semantic evaluation of body shapes in obesity surgery patients and overweight/obesity controls and found that both groups were more willing to accept positive adjectives as a match when BMI was low and negative adjectives as a match when BMI was high.	2	1	A recent paper has pointed out that post-bariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z). I think this aspect is interesting and linked to your results, showing a connection with clinical data.	1	2	obesities2010010_perova	1
+We have now added a Limitations and Future Research section to the Discussion to address this point and others: There are two limitations in our data curation process, the automated approach we used to check and select papers. Firstly, automatic classifiers of any sort can include some irrelevant or false positive articles. Due to the large amount of articles in our dataset, in Step 2 of our methodology, we developed a machine learning binary classifier – a support vector machine - with 87.56% accuracy to automatically identify relevant articles (accuracy is the number of correct predictions made, divided by the total number of predictions made, and then multiplied by 100 to convert it into a percentage). As a rule of thumb, accuracy of a predictive model that is above 80% is very commonly used to summarise the performance of that model. Still, it doesn’t exclude the possibility of irrelevant articles in our final dataset, but that possibility is less than 12.44 percent and this has to be considered against the benefit of scale and efficiency that this method allows. We also acknowledge that our analysis limited is in not being able to consider visual content visual content, images within articles (known as image framing), which have been shown to carry stigmatising elements [48]. This is something that could be added to the approach by including image classification along with additional measures. Furthermore, even though the Dow Jones is one the largest news databases, it might still miss some articles or news sources (although this doesn’t relate to the automatic approach as such). This applies to social media, even though this would not span as long a timeframe.	2	1	You report that an automatic approach was used to check all the papers. Is there any possible limit with this approach in the selection of the papers?	1	2	obesities2010010_perova	1
+Figures have been revised with addition of arrows and text to represent the data. Since a large number of ILs were screened, susceptible check (SC) was used with a set of ILs and hence SC couldn’t be shown in all figures. However, SC ‘HR12’ for blast disease was shown in all blast screening figures 3 to 5. Similarly, in Fig 5, ‘TN1’ and ‘Improved Samba Mahsuri’ as susceptible and resistant checks respectively for BB in comparison with IL-19031 were shown. Authors once again thank the reviewer for pointing out the mistake in legends. Now, we have rephrased the legends clearly describing all the terms.	2	1	The main issue with the ms is the quality of the figures: they are not clearly representing the data, arrows and text may help the reader, the controls are missing in most cases, and a general lack of precision is affecting them. Also, the legends are not clearly describing all the terms and should be revised. Legends should describe what is to be observed in the figure, possibly pointing to parts of special interest.	1	2	plants11050622_makarova	1
+Figure 1 legend has been revised with inclusion of the details of kharif and rabi seasons. kharif is the wet season with crop growing period from June to November and rabi is the dry season with crop growing period from December to May. We described kharif as wet season and rabi as dry season in the materials and methods section also.	2	1	Explaining briefly which and what are the indian sesasons during which experiments have been conducted would help clarity.	1	2	plants11050622_makarova	1
+Since the number of ILs is large, presenting phenotypic data for each IL will result in increasing the size of the main tables, hence data was earlier presented in supplementary tables. As suggested by the reviewers, we have revised the tables and presented the mean phenotypic data of BB, blast and drought screening in the main tables for each IL in parenthesis. Column on ‘no. of genes/QTL’ has been removed as suggested. Entry nos have been replaced with IL No as suggested.	2	1	"The phenotypic data are important, but currently they are presented only as additional material. The authors should consider reshaping the tables, eliminating unnecessary columns (e.g. no. of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion. Also ""entry nos."" is probably not clear and ILs no should maybe considered instead."	1	2	plants11050622_makarova	1
+The present work is not essentially a backcross breeding program aimed at development of near isogenic lines. However, Krishna Hamsa was the common background into which several genes/QTL were targeted from multiple donors and considering the morphological similarity between 27 ILs and Krishna Hamsa, background selection was done retrospectively. BGS validated our observations on morphological similarity. The same has been discussed in the 5th para under ‘discussion’. Also results on BGS have been presented under subsection 2.3 of results with data on polymorphic markers for BGS in supplementary table S8.	2	1	The data on bgs are not sufficiently presented in the text nor discussed.	1	2	plants11050622_makarova	1
+“()” have been removed while mentioned the numbers of the ILs and sentences have been revised appropriately in the manuscript.	2	1	"revise the use of ""-"" instead of ""( )"" when mentioning the numbers of the ILs, or any other system which would ensure a better homogeneity in comparison to the one present in the ms."	1	2	plants11050622_makarova	1
+The above mentioned lines have been checked and found either spelling mistakes or revision of sentences. Accordingly, corrections were made.	2	1	check lines 21, 37, 93, 153, 183-184, 312, 351-353, 360, 377, 466	1	2	plants11050622_makarova	1
+BLB has been replaced with BB throughout the manuscript.	2	1	stick to BB instead of BLB	1	2	plants11050622_makarova	1
+Legends of figures and tables and text in the results section have been revised with explanation of acronyms as suggested.	2	1	consider anticipating the explanation of the acronyms which are eventually presented only in the M&M, to facilitate the reading; consider this aspect also in the legends	1	2	plants11050622_makarova	1
+The sentence here is required to maintain flow of the subsequent content.	2	1	Lines 403 et seq. are redundant Response:	1	2	plants11050622_makarova	1
+Both lines explain our observations in different sets of ILs. Sentence at 351 explains susceptibility in ILs despite possessing the targeted gene/QTLs while sentence at 414 explains resistance in ILs despite the absence of targeted gene/QTLs.	2	1	lines 414 et seq. seem to be contradicting lines 351 et seq.	1	2	plants11050622_makarova	1
+The 85 ILs is a sum total of nine ILs marker positive to blast- R genes, nine ILs harbouring QTLs for drought tolerance and 67 ILs marker positive to BB-R genes mentioned at the beginning of 4th paragraph of discussion. As suggested, we have added in brief about the same at line 414.	2	1	the 85 ILs presented at 414 seem to be a bit out of the blue: a short intro to where they are coming from would help the reader.	1	2	plants11050622_makarova	1
+Authors profusely thank the reviewer for the appreciation.	2	1	Thank you for inviting me to review this manuscript. The paper itself is well written, although 1) somewhat results are partially descriptive and partially inferential. However, the authors have conducted a thorough literature review, undertaken a rigorous piece of data collection, and have generalized information accurately.	1	2	plants11050622_makarova	1
+Yes, we agree that there were some typo and spelling mistakes in the manuscript. The manuscript has been thoroughly revised for the same.	2	1	With minor grammatical revisions, the manuscript can be accepted as is.	1	2	plants11050622_makarova	1
+Authors feel greatly encouraged and motivated with the reviewer’s comments.	2	1	It is also acknowledged that this paper is probably the first of many papers to emerge from the study. As such, it is an overview paper that raises many questions. It would be interesting for the authors to provide more information about the research design and estimation methodologies, such as chi-square analysis of introgression lines, if possible for each crossed IL population. I only recommend some minor revisions before acceptance. Response: Authors once again thank the reviewer for correct assessment of the basic purpose of the manuscript. Yes, this is truly an overview paper presenting the interesting observations from our study. The introgression scheme and phenotyping of various biotic traits and drought phenotyping have been described in detail under ‘Materials and Methods’ section. The metric data on yield traits was statistically analyzed and results of ANOVA, heritability and critical differences have been presented in the manuscript. Authors agree that it would be more inferential with chi square values. However, the crossing scheme was viewed holistically and data on each cross was not maintained separately as the present study aimed at selecting introgression lines from multiples crosses with multiple stress resistance/tolerance by pooling several genes and QTLs into a common background. Despite maintaining large base populations, plants per se were selected based on marker positivity for inter-crossing and selfing and further stringent phenotypic selection for the targeted traits. Hence, chi-square which is perfectly apt for population derived from biparental crosses is not used in our study. This work is of outstanding quality, and I normally present more critical points in my reviews. However, this time it is just very beautiful work.	1	2	plants11050622_makarova	1
+‘=261%’ has been corrected to ‘+261%’ and typo error of ‘linkes’ corrected to ‘linked’	2	1	Line 262: “=261 %” , and Line 312 “linkes” a typo? Correct it Response:	1	2	plants11050622_makarova	1
+Thank you for the positive comments.	2	1	It was a pleasure to read this manuscript. I wish the author of the best.	1	2	plants11050622_makarova	1
+We appreciate the suggestions from the reviewer, which has resulted in improving the message of the manuscript.	2	1	This paper described the multi-parents introgression assisted by molecular markers. The content is informative, however, the tables should be reorganized and the statistical methods description have to be improved. Please see the suggestion and comments below.	1	2	plants11050622_makarova	1
+Added references appropriately at two places as suggested by the reviewer.	2	1	L48-63: should add some citations on the first part of introduction.	1	2	plants11050622_makarova	1
+Explanation for boro season has been added.	2	1	L127: Please add the explanation of “boro season”.	1	2	plants11050622_makarova	1
+Legends of the supplementary tables have been revised and inference of the table is given in foot note.	2	1	The legend of supplementary table should be improved. Table and main text are independent, so the authors have to describe the table more carefully.	1	2	plants11050622_makarova	1
+Authors once again thank the reviewer for the valuable suggestion. More details on the statistical analysis have been added as suggested.	2	1	L562: the section of statistical analysis should add more details. Also please indicate the R version. For example, how does H2 calculate?	1	2	plants11050622_makarova	1
+Yes, significant means p-value. In the supplementary tables, expanded form of DFF is given as suggested. Treatments refers to introgression lines and check to control. For uniformity, we have changed the terminology to treatment instead of using IL or variety and check to control in all the revised supplementary tables and rephrased the legends accordingly.	2	1	On Supplementary table S2, does significant mean p-value? Indicate DFF = days to fifty percent flowering. What is the “treatment”? What do the results on “check” mean? How did you analyze “control vs IL”?	1	2	plants11050622_makarova	1
+Similar to Supplementary Table S2, S3 has been revised	2	1	On Supplementary table S3, similar questions as S2, please also explain.	1	2	plants11050622_makarova	1
+CD is the critical difference at 1% and 5% level of significance (p-value) for testing of significant differences among the ILs. The details of CD calculation have been added in the materials and methods section as suggested.	2	1	On Supplementary table S5, what is “C.D”? How did you calculate those values on those comparison types?	1	2	plants11050622_makarova	1
+Supplementary table S5 is on CD and corrections have been addressed as suggested as at S. No 7. Supplementary table S6 have been modified by shifting data of BB and blast scores to main table. Each PC group is mentioned on top as sub heading at the start of each group. Each IL is presented only once in the entire table under separate PC groups.	2	1	Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups. Then the table can be more informative and ease to read.	1	2	plants11050622_makarova	1
+Full names of the abbreviations have been added as suggested.	2	1	Please add the full name of abbreviations. For example on L203 “CD”, L211 “SES“, L228 “UBN“ and L332 “ICAR-IRRR”.	1	2	plants11050622_makarova	1
+Subtitle has been added as suggested.	2	1	The last paragraph of results is about background selection. Suggest to add subtitle “2.2.8”.	1	2	plants11050622_makarova	1
+A representative gel picture has been added as suggested.	2	1	Please add some gel pictures of the foreground selection markers you used in order to visualize the genotyping results and showed the polymorphism of these markers on gel.	1	2	plants11050622_makarova	1
+Table S8 represents polymorphic markers between pairs of parents including some common polymorphic markers. Hence the total number represented in the table S8 is not additive and not matching with the numbers given in the text. 124 is the total polymorphic markers excluding repetition. 27 ILs were selected based on their agro-morphological similarity with recurrent parent ‘Krishna Hamsa’ and evaluated for background recovery. The list of 27 ILs has been included in the revised manuscript as suggested.	2	1	L296-L302: the marker descriptions on main text cannot match the supplementary table S8. Also, what are those 27 ILs on L298?	1	2	plants11050622_makarova	1
+Mention of appendix at L602 is a typo error and has been removed	2	1	L602: there is no appendix.	1	2	plants11050622_makarova	1
+We agree with your comments. The term ‘expression’ has, therefore, been changed to ‘abundance’ or ‘level’ throughout the manuscript.	2	1	In general, the manuscript refers often to PGRMC1 expression or downregulation. These are fuzzy terms for proteins. It is much preferable to refer to protein abundance, and changes in abundance. Such changes may occur at the translational or post-translational level. Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool. The authors are strongly advised to change from e.g. ‘expression’ to ‘abundance’ throughout.	1	2	reprodmed3020015_makarova	1
+All experiments were repeated at least two times. We have failed the description regarding the reproducible results in the previously submitted manuscript, and so revised the section of the methods in the revised manuscript (Page 4, lins143-143).	2	1	Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented. Therefore, the reviewer assumes that experiments have only been performed once. The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.	1	2	reprodmed3020015_makarova	1
+We agree that AG-205 is not a specific inhibitor of PGRMC1. Therefore, we examined the effects of siRNA-mediated PGRMC1 knockdown on decidualization. As you mentioned, this issue was not appropriately described in the previously submitted manuscript. The citations have been inserted in the revised manuscript (page 8, lines 258-263).	2	1	Line 24. AG-205. The authors must refer to the fact that AG-205 is not specific for PGRMC1, contrary to many citations in the literature (such as would be the case here, if uncorrected). Cite PMID: 34944026, PMID: 34680104, PMID: 32924377. Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.	1	2	reprodmed3020015_makarova	1
+The name of the reagent has been corrected to AG-205 consistently throughout the manuscript.	2	1	It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).	1	2	reprodmed3020015_makarova	1
+The sentence has been separated according to your suggestion.	2	1	Line 50. “(10-12); however,” Two discrete sentences “(10-12).	1	2	reprodmed3020015_makarova	1
+The description has been modified as followed (page 2, line 69). ‘Endometrial samples were obtained from Japanese patients with a normal menstrual cycle, ….’.	2	1	Lines 76-78. Statistical sample sizes were unfavourably small. The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).	1	2	reprodmed3020015_makarova	1
+The primary cultured ESCs we used were confirmed immunocytochemically to be positive for vimentin, a marker for stromal cells, and negative for cytokeratin, a marker for epithelial cells (please see above figure). ESCs at early passages (between 2 and 6 passages) were used for the experiment. In this study, the passaged cells were used without freezing. However, we have confirmed that primary ESCs can be cryopreserved in CELLBANKER, and retain characteristics such as responsiveness to decidual stimuli. These descriptions have been added to the revised manuscript (page 3, lines 96-99). Figure. Immunocytochemistry (ICC) of vimentin and cytokeratin in isolated human ESCs.	2	1	Lines 88-98. Cell culture. What characterization was performed to confirm the identity of cells as ESCs? In general, there is very little detail on these primary cultured lines. How long are they held in culture? Are they stored cryogenically? How are they re-seeded into culture?	1	2	reprodmed3020015_makarova	1
+The protein samples were separated by SDS-PAGE (5-20% gradient gel) and transferred electrophoretically to polyvinylidene difluoride membranes (8 cm x 8.5 cm) for 60 min at constant current of 128 mA using a semi-dry transfer system (ATTO, Tokyo Japan). These conditions have been added to the Materials and Methods section (Pages 3 and 4, lines 127-130).	2	1	Lines 121-133. What percentage acrylamide were the SDS PAGE gels. For what length of time and at what voltage settings was electrophoretic transfer to PVDF? In what device?	1	2	reprodmed3020015_makarova	1
+The catalogue number of the antibodies (PI-1000, PI-2000) were inserted (Page 4, line 134).	2	1	Line 126-7. Give the catalogue number of the secondary antibody.	1	2	reprodmed3020015_makarova	1
+The immunoreactive bands were visualized using an enhanced chemiluminescence (Western Lightning, PerkinElmer, Inc., Waltham, MA, USA) and analyzed with ImageQuant LAS 500 (semi-auto mode; GE Healthcare Japan, Tokyo Japan) (Page 4, lines 136-137).	2	1	Line 131. On what device was ECL light detected? What were the device settings? How was pixel saturation dealt with?	1	2	reprodmed3020015_makarova	1
+The densitometry analysis was carried out using the “Gel Plot” plug-in of the Image J software. This plug-in has been described in the Materials and Methods (Page 4, line 139) 12)	2	1	Line 13 Which ImageJ plug-in was used for quantification?	1	2	reprodmed3020015_makarova	1
+As you indicated, all data are expressed consistently as means and standard deviation (Figs 1 to 4). In our previously submitted manuscript, we had described carelessly the data of as for figure 5B and C with s.d.	2	1	Lines 134-137. Result errors should not be expressed as SEM, but as standard deviation (s.d.). All results should show s.d. error bars.	1	2	reprodmed3020015_makarova	1
+Thank you for pointing this out. The mistakes have been corrected (Pages 4,5 and 7, lines 160, 174 and 217).	2	1	Line 150. The legend to fig. 1. inappropriately ends with “2.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.	1	2	reprodmed3020015_makarova	1
+Based upon your suggestions, additional experiments have been performed and added individual Western blotting data from there different ESCs (named #1-3) in Figure 2.	2	1	Lines 153-156, Figure 2. The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient. This result must be demonstrated in a substantively convincing manner.	1	2	reprodmed3020015_makarova	1
+Western blotting for PGRMC1 in figures 2, 3, and 4 have been replaced in the data with the whole molecular weight range. As you pointed out, PGRMC1 is known to be modified by sumoylation or ubiquitinylation which results in high molecular weight. The right panel shows the western blotting of PGRMC1. There are some higher molecular bands (white arrowhead) in lysates of ESCs, however, the band’s intensity was not changed by PGRMC1 siRNA treatment. Therefore, we believe that these high molecular bands detected in our experiment were non-specific bands.	2	1	2. Line 158/Fig. The western blot (also Fig.3) result must show the entire molecular weight range (e.g. in supplemental data), including MW markers. PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491). The latter could well contribute to alterations in PGRMC1 abundance cited by this study. It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.	1	2	reprodmed3020015_makarova	1
+As you pointed out, the possible involvement of posttranslational modification of endometrial PGRMC1 such as sumoylation and ubiquitinylation is interesting. Discussion regarding this possibility has been inserted in the revised manuscript (Page 9, lines 300-307). We will further examine whether posttranslational modification of PGRMC1 could be associated with decidualization in the next study.	2	1	Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true). Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels. In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization. It is important to determine whether these mechanisms are both operating. At the very least this mechanism merits detailed discussion in that section. PGRMC1 protein instability could be the dominant effect involved, with miRNA transcript regulation playing only a minor part.	1	2	reprodmed3020015_makarova	1
+The mistake has been corrected.	2	1	Line 163. Change 3.2. to 3.3.	1	2	reprodmed3020015_makarova	1
+All data of cell culture represent the mean ± SD of three independent experiments with at least repeated two times each experiment. The explanation of the experiments has been inserted the method (page 4, lines142-143).	2	1	Lines 164-172. This cell culture experiment also seems to be based upon just one primary cell culture, split into replicates. I am uncertain because the methods do not explain how the experiment was performed. Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.	1	2	reprodmed3020015_makarova	1
+Thank you for your valuable advice to prove the significance of PGRMC1 downregulation in decidualization. Salsano et al. (19), reported that overexpression of PGRMC1 in ESCs abrogated the decidual markers expression (page 8. Lines 253-254). We will try to examine the effects of enforced expression of codon-altered PGRMC1 which is resistant for siRNA on decidualization in next study.	2	1	Lines 178-9. If PGRMC1 down-regulation is critical, it should be possible to perturb decidualization by expression of an exogenous PGRMC1 protein. This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA. If siRNA-resistant PGRMC1 levels are able to inhibit decidualization it would provide convincing evidence in favour of the hypothesis that lowered PGRMC1 is required for decidualization, and is perhaps even a trigger. A codon-altered PGRMC1 expression vector would cost just several hundred dollars from a provider such as Genscript, so the experiment is eminently achievable.	1	2	reprodmed3020015_makarova	1
+Based upon your suggestions, we have added the data of PGRMC1 knockdown in figure 4. We confirmed efficient siRNA-mediate PGRMC1 knockdown by western blotting.	2	1	Notably, Fig. 4. lacks the western blot equivalent to Fig.3A, showing that siRNA indeed depleted PGRMC1 levels.	1	2	reprodmed3020015_makarova	1
+The statistical analysis was not appropriate in the former manuscript. The data has been re-evaluated by a two-way ANOVA followed by a Turkey-Kramer multiple comparison test (page 4, line13).	2	1	Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test. Assuming binomial distribution, a two-way ANOVA is required for these experiments.	1	2	reprodmed3020015_makarova	1
+As you suggested, we should have presented the data in the same graph. However, the two experiments were performed independently. Therefore, we were not able to combine the results in the graph. Because treatment with P4 alone could not induce decidualization in the experiments, we examined the effect of PGRMC1 knockdown and inhibition on db-cAMP-induced decidualization. Further study would be needed to determine whether PGRMC1 is involved in the P4 action in endometrium.	2	1	Line 189-96. The paragraph describes a result showing that the results are independent of P4. However, the control structure of comparisons is incorrect. The simplest direct comparison would be db-cAMP/P4 vs. db-cAMP to permit the argument the authors are presenting. However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4. The statistical significance of any result would be evaluated using multivariate analysis.	1	2	reprodmed3020015_makarova	1
+The description has been corrected and cited the references in the section (page 7, lines 223-224).	2	1	Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1. The results of PMID: 22492871 and PMID: 21109987 MUST be cited here. This is not a novel finding.	1	2	reprodmed3020015_makarova	1
+New discussion regarding the need for PGRMC1 knockout studies to clarify the role of PGRMC1 in decidualization has been inserted in page 9,	2	1	The discussion must qualify the results of Salsano et al. who used AG-205 to inhibit PGRMC1. There was no control in PGRMC1 genomic/CRISPR KO cells (not si/miRNA KO) to demonstrate that effects induced by AG-205 were due to PGRMC1.	1	2	reprodmed3020015_makarova	1
+The error has been corrected.	2	1	Line 261. mechanisms HAVE not (plural mechanisms)	1	2	reprodmed3020015_makarova	1
+Your correction is what we meant to mention. We have replaced this sentence in the revised manuscript. (page9, line 284-286) 27)	2	1	Lines 261-262. “In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded. Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) >>	1	2	reprodmed3020015_makarova	1
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, line 295-299).	2	1	Lines 271-76. This is very long and burdens the reader. Always make new sentences when possible. Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy. Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation. Furthermore, controlling their expression may support the improvement of IVF (NOT IFV) outcome. Define IVF at first use.	1	2	reprodmed3020015_makarova	1
+The report that PGRMC1 mediates H2O2-induced cell death in MCF7 cells has been inserted in discussion (page 10, lines 314-317)	2	1	Lines 281-5. PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988). That could be discussed here in terms of mechanism.	1	2	reprodmed3020015_makarova	1
+We have suggested the possibility that PGRMC1 downregulation may be involved in ESCs senescence during the decidualization (page 10, lines 319-321).	2	1	Lines 286-287. Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?). Suggest observations that may be able to discriminate between different possibilities.	1	2	reprodmed3020015_makarova	1
+As you pointed out, we infer that downregulation of PGRMC1 may be necessary for decidualization to proceed. To clarify this point, we further attempted to overexpress PGRMC1 by introducing expression vectors into primary ESCs under various conditions using lipofection and calcium phosphate transfection methods, as you suggested. Unfortunately, for reasons unknown, we were unable to establish ESCs overexpressing PGRMC1. In the present study, we found that not only knockdown of PGRMC1 but also functional inhibition with inhibitors promotes ESC decidualization in vitro. Furthermore, in a previous report, overexpression of PGRMC1 suppressed ESCs differentiation (Ref. 19). These results indicate that decreased PGRMC1 in the endometrium may be associated with accelerated dedifferentiation. As you suggested, we have changed the title significantly because further evidence was not available. We plan to establish a model of miR-resistant PGRMC1 overexpression using several ESCs cell lines to investigate the role of PGRMC1 in decidualization, and if possible, we would be happy to make this our next research topic.	4	1	Furthermore, the required experiment is technically not challenging. The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed. The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required. This may be only correlation.	3	2	reprodmed3020015_makarova	1
+Thank you for correcting the sentence. The description has been corrected in accordance with your suggestion (page 4, lines 141-142).	4	1	Lines 142-3. “Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments. All reported results were observed in at least two independent experiments.	3	2	reprodmed3020015_makarova	1
+The error has been corrected (page 8, line 257).	4	1	Line 259. PGRMC1-independe (add t) >>	3	2	reprodmed3020015_makarova	1
+The description has been modified to the following (page 8, lines 261-266). “Although we cannot exclude the possible non-specific action of AG-205 on ESCs, the effect of AG-205 treatment and PGRMC1 knockdown promoted in vitro decidualization. These findings suggested that PGRMC1 downregulation may promote ESC decidualization during the secretory phase.	4	1	Lines 263-267: “Because, the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence. What is being said?	3	2	reprodmed3020015_makarova	1
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, lines 280-281).	4	1	lines 281-282. There are two natural sentences here. However, as described above AG-205 is not a specific inhibitor of PGRMC1. [full stop, new sentence] Further study of PGRMC1 knockout on decidualization is needed.	3	2	reprodmed3020015_makarova	1
+As you pointed out, we did not explain the relationship between PGRMC1 stability and post-transcriptional regulation of the protein. We have added the following description (page 9, lines 304-308). ”These reports suggested that PGRMC1 protein levels can be modulated by changes in the rates of transcription, translation, and degradation. Therefore, further studies would be needed to clarify the possible involvement of the post-translational modification and degradation that may regulate PGRMC1 protein stability in ESCs during decidualization.”	4	1	The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation. Sabbir’s results (ref 31) imply that this also holds for PGRMC1. The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned. To be clear, protein levels can be mediated by altered rates of transcription, translation, or degradation. This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues. Lines 300-307.	3	2	reprodmed3020015_makarova	1
+We appreciate your review of our revised manuscript, constructive comments, and suggestions for improving the manuscript. We deeply understand the significance of the experiments using miR-98-resistant PGRMC stably expressing ESCs to directly prove the implication of the miR-mediated PGRMC1 regulation in decidualization. We are transfecting the miR-resistant PGRMC1 expression vector into ESCs cell lines to explore the role of PGRMC1 in decidualization but is not going well. As you advised, lentiviral transduction of the PGRMC1 along with fluorescent protein may enable us to effectively select the overexpressing cells without losing differentiation status in response to decidual stimuli. We have acknowledged this and suggested it as a topic for further research in the “Limitation” section of the revised manuscript following editor’s comment (page 10, lines 336-348). “Limitations: In this study, we proposed that miR-98-mediated PGRMC1 downregulation may be involved in decidualization, but were unable to examine the effects of miR-98-resistant PGRMC1 on differentiation due to technical difficulties establishing PGRMC1-overexpressing ESCs. miR-98 may not be the only miRNA that modulates PGRMC1 expression; therefore, comprehensive analysis of transcriptional, post-transcriptional, and post-translational regulation of endometrial PGRMC1 during decidualization is required. In addition, PGRMC1 can bind to various proteins, including receptors for epidermal growth factor [46] and insulin [47], cytochrome P450 [48], and serpine mRNA-binding protein 1 (SERBP1) [19]. Further, PGRMC1 may interact with proteins associated with endomembrane trafficking/cytoskeleton and mitochondrial functions in decidualizing ESCs [49]. Thus, we plan to investigate the modulation of endometrial PGRMC1 expression and the interaction between PGRMC1 and intracellular proteins during decidualization in the future.”	6	1	In conclusion, if this published was published in present form then it would invariably be cited as demonstrating that PGRMC1 downregulation by miR-98 causes decidualization. Since that is by no means certain, the study should not be published in its present form. The opportunity should remain open for the authors to convincingly demonstrate the requirement for PGRMC1 down-regulation in decidualization by performing the requested experiments, or others of their devising. However, in the first instance the manuscript cannot be published as it stands, and should therefore be rejected now with the stated option of subsequent resubmission only if new evidence is obtained.	5	2	reprodmed3020015_makarova	1
+Our responses to your comments are as follows. We appreciate your review of our revised manuscript, constructive comments, and suggestions. As you pointed out, we have deleted the proposed model of the study (Figure 6). We have added the “Strength and limitation of this study” and “Conclusions” in the revised manuscript in accordance with the academic editor’s comments.	8	1	The reviewer is not convinced that PGRMC1 level attenuation by miR-98 mechanistically drives attenuation. Cell biology is quite complex, and it remains fully feasible that PGRMC1 levels only correlate with the decidualization process. In the lack of convincing evidence, the manuscript should be rejected.	7	2	reprodmed3020015_makarova	1
+As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？The sample size is small. And please provide basic information of the patient.	1	2	reprodmed3020015_makarova	1
+Because of the ethical difficulties of isolating ESCs from disease-free healthy women, we routinely used ESCs isolated from the patients in surgical cases. ESC lines are commercially available (T-HESC, ATCC) and maybe useful tool to investigate the role of PGRMC1 in decidualization, as you pointed out. However, the cell lines are also established from the endometrium of leiomyoma and immortalized by transfection with hTERT. Therefore, we believe that ESCs used in this study are no different than the above cells and cell lines in examining the role of PGRMC1.In the future, we plan to use several ESCs and their cell lines to clarify the precise mechanisms of PGRMC1 downregulation and decidualization.	2	1	The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough. It is suggested to add cell lines or normal endometrial cells for verification. If further mechanism exploration is carried out, the research will be more complete and persuasive.	1	2	reprodmed3020015_makarova	1
+We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript. We are thankful for the time and energy you expended.	4	1	The author provided explanations for the questions. The manuscript has been modified accordingly. I think the manuscript could be accepted now.	3	2	reprodmed3020015_makarova	1
+The author provided explanations for the questions. The manuscript has been modified accordingly. I think the manuscript could be accepted now.	3	2	As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	reprodmed3020015_makarova	1
+We agree with your comments. The term ‘expression’ has, therefore, been changed to ‘abundance’ or ‘level’ throughout the manuscript.	2	1	1) In general, the manuscript refers often to PGRMC1 expression or downregulation. These are fuzzy terms for proteins. It is much preferable to refer to protein abundance, and changes in abundance. Such changes may occur at the translational or post-translational level. Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool. The authors are strongly advised to change from e.g. ‘expression’ to ‘abundance’ throughout.	1	2	reprodmed3020015_perova	1
+All experiments were repeated at least two times. We have failed the description regarding the reproducible results in the previously submitted manuscript, and so revised the section of the methods in the revised manuscript (Page 4, lins143-143).	2	1	2) Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented. Therefore, the reviewer assumes that experiments have only been performed once. The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.	1	2	reprodmed3020015_perova	1
+We agree that AG-205 is not a specific inhibitor of PGRMC1. Therefore, we examined the effects of siRNA-mediated PGRMC1 knockdown on decidualization. As you mentioned, this issue was not appropriately described in the previously submitted manuscript. The citations have been inserted in the revised manuscript (page 8, lines 258-263).	2	1	3) Line 24. AG-205. The authors must refer to the fact that AG-205 is not specific for PGRMC1, contrary to many citations in the literature (such as would be the case here, if uncorrected). Cite PMID: 34944026, PMID: 34680104, PMID: 32924377. Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.	1	2	reprodmed3020015_perova	1
+The name of the reagent has been corrected to AG-205 consistently throughout the manuscript.	2	1	4) It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).	1	2	reprodmed3020015_perova	1
+The sentence has been separated according to your suggestion.	2	1	5) Line 50. “(10-12); however,” Two discrete sentences “(10-12).	1	2	reprodmed3020015_perova	1
+The description has been modified as followed (page 2, line 69). ‘Endometrial samples were obtained from Japanese patients with a normal menstrual cycle, ….’.	2	1	6) Lines 76-78. Statistical sample sizes were unfavourably small. The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).	1	2	reprodmed3020015_perova	1
+The primary cultured ESCs we used were confirmed immunocytochemically to be positive for vimentin, a marker for stromal cells, and negative for cytokeratin, a marker for epithelial cells (please see above figure). ESCs at early passages (between 2 and 6 passages) were used for the experiment. In this study, the passaged cells were used without freezing. However, we have confirmed that primary ESCs can be cryopreserved in CELLBANKER, and retain characteristics such as responsiveness to decidual stimuli. These descriptions have been added to the revised manuscript (page 3, lines 96-99).	2	1	7) Lines 88-98. Cell culture. What characterization was performed to confirm the identity of cells as ESCs? In general, there is very little detail on these primary cultured lines. How long are they held in culture? Are they stored cryogenically? How are they re-seeded into culture?	1	2	reprodmed3020015_perova	1
+The protein samples were separated by SDS-PAGE (5-20% gradient gel) and transferred electrophoretically to polyvinylidene difluoride membranes (8 cm x 8.5 cm) for 60 min at constant current of 128 mA using a semi-dry transfer system (ATTO, Tokyo Japan). These conditions have been added to the Materials and Methods section (Pages 3 and 4, lines 127-130).	2	1	8) Lines 121-133. What percentage acrylamide were the SDS PAGE gels. For what length of time and at what voltage settings was electrophoretic transfer to PVDF? In what device?	1	2	reprodmed3020015_perova	1
+The catalogue number of the antibodies (PI-1000, PI-2000) were inserted (Page 4, line 134).	2	1	9) Line 126-7. Give the catalogue number of the secondary antibody.	1	2	reprodmed3020015_perova	1
+The immunoreactive bands were visualized using an enhanced chemiluminescence (Western Lightning, PerkinElmer, Inc., Waltham, MA, USA) and analyzed with ImageQuant LAS 500 (semi-auto mode; GE Healthcare Japan, Tokyo Japan) (Page 4, lines 136-137).	2	1	10) Line 131. On what device was ECL light detected? What were the device settings? How was pixel saturation dealt with?	1	2	reprodmed3020015_perova	1
+The densitometry analysis was carried out using the “Gel Plot” plug-in of the Image J software. This plug-in has been described in the Materials and Methods (Page 4, line 139)	2	1	11) Line 13 Which ImageJ plug-in was used for quantification?	1	2	reprodmed3020015_perova	1
+As you indicated, all data are expressed consistently as means and standard deviation (Figs 1 to 4). In our previously submitted manuscript, we had described carelessly the data of as for figure 5B and C with s.d.	2	1	12) Lines 134-137. Result errors should not be expressed as SEM, but as standard deviation (s.d.). All results should show s.d. error bars.	1	2	reprodmed3020015_perova	1
+Thank you for pointing this out. The mistakes have been corrected (Pages 4,5 and 7, lines 160, 174 and 217).	2	1	13) Line 150. The legend to fig. 1. inappropriately ends with “2.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2.	1	2	reprodmed3020015_perova	1
+Based upon your suggestions, additional experiments have been performed and added individual Western blotting data from there different ESCs (named #1-3) in Figure 2.	2	1	14) Lines 153-156, Figure 2. The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient. This result must be demonstrated in a substantively convincing manner.	1	2	reprodmed3020015_perova	1
+Western blotting for PGRMC1 in figures 2, 3, and 4 have been replaced in the data with the whole molecular weight range. As you pointed out, PGRMC1 is known to be modified by sumoylation or ubiquitinylation which results in high molecular weight. The right panel shows the western blotting of PGRMC1. There are some higher molecular bands (white arrowhead) in lysates of ESCs, however, the band’s intensity was not changed by PGRMC1 siRNA treatment. Therefore, we believe that these high molecular bands detected in our experiment were non-specific bands.	2	1	2. 15) Line 158/Fig. The western blot (also Fig.3) result must show the entire molecular weight range (e.g. in supplemental data), including MW markers. PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491). The latter could well contribute to alterations in PGRMC1 abundance cited by this study. It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.	1	2	reprodmed3020015_perova	1
+As you pointed out, the possible involvement of posttranslational modification of endometrial PGRMC1 such as sumoylation and ubiquitinylation is interesting. Discussion regarding this possibility has been inserted in the revised manuscript (Page 9, lines 300-307). We will further examine whether posttranslational modification of PGRMC1 could be associated with decidualization in the next study.	2	1	16) Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true).	1	2	reprodmed3020015_perova	1
+The mistake has been corrected.	2	1	17) Line 163. Change 3.2. to 3.3.	1	2	reprodmed3020015_perova	1
+All data of cell culture represent the mean ± SD of three independent experiments with at least repeated two times each experiment. The explanation of the experiments has been inserted the method (page 4, lines142-143).	2	1	18) Lines 164-172. This cell culture experiment also seems to be based upon just one primary cell culture, split into replicates. I am uncertain because the methods do not explain how the experiment was performed. Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.	1	2	reprodmed3020015_perova	1
+Thank you for your valuable advice to prove the significance of PGRMC1 downregulation in decidualization. Salsano et al. (19), reported that overexpression of PGRMC1 in ESCs abrogated the decidual markers expression (page 8. Lines 253-254). We will try to examine the effects of enforced expression of codon-altered PGRMC1 which is resistant for siRNA on decidualization in next study.	2	1	19) Lines 178-9. If PGRMC1 down-regulation is critical, it should be possible to perturb decidualization by expression of an exogenous PGRMC1 protein. This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA. If siRNA-resistant PGRMC1 levels are able to inhibit decidualization it would provide convincing evidence in favour of the hypothesis that lowered PGRMC1 is required for decidualization, and is perhaps even a trigger. A codon-altered PGRMC1 expression vector would cost just several hundred dollars from a provider such as Genscript, so the experiment is eminently achievable.	1	2	reprodmed3020015_perova	1
+Based upon your suggestions, we have added the data of PGRMC1 knockdown in figure 4. We confirmed efficient siRNA-mediate PGRMC1 knockdown by western blotting.	2	1	20) Notably, Fig. 4. lacks the western blot equivalent to Fig.3A, showing that siRNA indeed depleted PGRMC1 levels.	1	2	reprodmed3020015_perova	1
+The statistical analysis was not appropriate in the former manuscript. The data has been re-evaluated by a two-way ANOVA followed by a Turkey-Kramer multiple comparison test (page 4, line13).	2	1	21) Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test. Assuming binomial distribution, a two-way ANOVA is required for these experiments.	1	2	reprodmed3020015_perova	1
+As you suggested, we should have presented the data in the same graph. However, the two experiments were performed independently. Therefore, we were not able to combine the results in the graph. Because treatment with P4 alone could not induce decidualization in the experiments, we examined the effect of PGRMC1 knockdown and inhibition on db-cAMP-induced decidualization. Further study would be needed to determine whether PGRMC1 is involved in the P4 action in endometrium.	2	1	22) Line 189-96. The paragraph describes a result showing that the results are independent of P4. However, the control structure of comparisons is incorrect. The simplest direct comparison would be db-cAMP/P4 vs. db-cAMP to permit the argument the authors are presenting. However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4. The statistical significance of any result would be evaluated using multivariate analysis.	1	2	reprodmed3020015_perova	1
+The description has been corrected and cited the references in the section (page 7, lines 223-224).	2	1	23) Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1. The results of PMID: 22492871 and PMID: 21109987 MUST be cited here. This is not a novel finding.	1	2	reprodmed3020015_perova	1
+New discussion regarding the need for PGRMC1 knockout studies to clarify the role of PGRMC1 in decidualization has been inserted in page 9, lines 281-282.	2	1	24) The discussion must qualify the results of Salsano et al. who used AG-205 to inhibit PGRMC1. There was no control in PGRMC1 genomic/CRISPR KO cells (not si/miRNA KO) to demonstrate that effects induced by AG-205 were due to PGRMC1.	1	2	reprodmed3020015_perova	1
+The error has been corrected.	2	1	25) Line 261. mechanisms HAVE not (plural mechanisms) >>	1	2	reprodmed3020015_perova	1
+Your correction is what we meant to mention. We have replaced this sentence in the revised manuscript. (page9, line 284-286)	2	1	26) Lines 261-262. “In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded. Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) >>	1	2	reprodmed3020015_perova	1
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, line 295-299).	2	1	27) Lines 271-76. This is very long and burdens the reader. Always make new sentences when possible. Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy. Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation. Furthermore, controlling their expression may support the improvement of IVF (NOT IFV) outcome. Define IVF at first use.	1	2	reprodmed3020015_perova	1
+The report that PGRMC1 mediates H2O2-induced cell death in MCF7 cells has been inserted in discussion (page 10, lines 314-317)	2	1	28) Lines 281-5. PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988). That could be discussed here in terms of mechanism.	1	2	reprodmed3020015_perova	1
+We have suggested the possibility that PGRMC1 downregulation may be involved in ESCs senescence during the decidualization (page 10, lines 319-321).	2	1	29) Lines 286-287. Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?). Suggest observations that may be able to discriminate between different possibilities.	1	2	reprodmed3020015_perova	1
+We appreciate your review of our revised manuscript, constructive comments, and suggestions for improving the manuscript. We deeply understand the significance of the experiments using miR-98-resistant PGRMC stably expressing ESCs to directly prove the implication of the miR-mediated PGRMC1 regulation in decidualization. We are transfecting the miR-resistant PGRMC1 expression vector into ESCs cell lines to explore the role of PGRMC1 in decidualization but is not going well. As you advised, lentiviral transduction of the PGRMC1 along with fluorescent protein may enable us to effectively select the overexpressing cells without losing differentiation status in response to decidual stimuli. We have acknowledged this and suggested it as a topic for further research in the “Limitation” section of the revised manuscript following editor’s comment (page 10, lines 336-348). “Limitations: In this study, we proposed that miR-98-mediated PGRMC1 downregulation may be involved in decidualization, but were unable to examine the effects of miR-98-resistant PGRMC1 on differentiation due to technical difficulties establishing PGRMC1-overexpressing ESCs. miR-98 may not be the only miRNA that modulates PGRMC1 expression; therefore, comprehensive analysis of transcriptional, post-transcriptional, and post-translational regulation of endometrial PGRMC1 during decidualization is required. In addition, PGRMC1 can bind to various proteins, including receptors for epidermal growth factor [46] and insulin [47], cytochrome P450 [48], and serpine mRNA-binding protein 1 (SERBP1) [19]. Further, PGRMC1 may interact with proteins associated with endomembrane trafficking/cytoskeleton and mitochondrial functions in decidualizing ESCs [49]. Thus, we plan to investigate the modulation of endometrial PGRMC1 expression and the interaction between PGRMC1 and intracellular proteins during decidualization in the future.”	4	1	In response the authors have made minimal changes to the manuscript, effectively restricted to rewording the title, and several minor text changes. Unfortunately, the authors have been unable to demonstrate the direct involvement of PGRMC1 downregulation in decidualization. Since this is the main publishable finding, the results as they stand do not merit publication. The authors mention technical difficulties in establishing ESCs that express miRNA-resistant PGRMC1. This may be due to stable plasmid selection conditions, which may affect the differentiation status of the ESC cells. It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1. An inducible vector may be advisable, such as tetracycline, IPTG, or other available systems. Such vectors typically include fluorescent protein reporters, enabling discrimination between infected and non-infected cells in one plate, or population separation by FACS.	3	2	reprodmed3020015_perova	1
+Our responses to your comments are as follows. We appreciate your review of our revised manuscript, constructive comments, and suggestions. As you pointed out, we have deleted the proposed model of the study (Figure 6). We have added the “Strength and limitation of this study” and “Conclusions” in the revised manuscript in accordance with the academic editor’s comments.	6	1	The inclusion of the model of Fig. 6 in the manuscript depicts the problem. This manuscript will be cited as evidence of that inconclusive model. The reviewer is not convinced that PGRMC1 level attenuation by miR-98 mechanistically drives attenuation. Cell biology is quite complex, and it remains fully feasible that PGRMC1 levels only correlate with the decidualization process. In the lack of convincing evidence, the manuscript should be rejected.	5	2	reprodmed3020015_perova	1
+As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？	1	2	reprodmed3020015_perova	1
+Because of the ethical difficulties of isolating ESCs from disease-free healthy women, we routinely used ESCs isolated from the patients in surgical cases. ESC lines are commercially available (T-HESC, ATCC) and maybe useful tool to investigate the role of PGRMC1 in decidualization, as you pointed out. However, the cell lines are also established from the endometrium of leiomyoma and immortalized by transfection with hTERT. Therefore, we believe that ESCs used in this study are no different than the above cells and cell lines in examining the role of PGRMC1.In the future, we plan to use several ESCs and their cell lines to clarify the precise mechanisms of PGRMC1 downregulation and decidualization.	2	1	The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough. It is suggested to add cell lines or normal endometrial cells for verification. If further mechanism exploration is carried out, the research will be more complete and persuasive.	1	2	reprodmed3020015_perova	1
+As you pointed out, we infer that downregulation of PGRMC1 may be necessary for decidualization to proceed. To clarify this point, we further attempted to overexpress PGRMC1 by introducing expression vectors into primary ESCs under various conditions using lipofection and calcium phosphate transfection methods, as you suggested. Unfortunately, for reasons unknown, we were unable to establish ESCs overexpressing PGRMC1. In the present study, we found that not only knockdown of PGRMC1 but also functional inhibition with inhibitors promotes ESC decidualization in vitro. Furthermore, in a previous report, overexpression of PGRMC1 suppressed ESCs differentiation (Ref. 19). These results indicate that decreased PGRMC1 in the endometrium may be associated with accelerated dedifferentiation. As you suggested, we have changed the title significantly because further evidence was not available. We plan to establish a model of miR-resistant PGRMC1 overexpression using several ESCs cell lines to investigate the role of PGRMC1 in decidualization, and if possible, we would be happy to make this our next research topic.	2	1	I requested a major revision including the use of miRNA-resistant exogenous PGRMC1 expression (altered codon usage) to demonstrate that lack of PGRMC1 attenuation will prevent decidualization. That new experimental result is what elevated the required changes to a major revision. The revised manuscript as presently resubmitted is a minor revision. This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization. This requires stronger evidence than that currently presented. Furthermore, the required experiment is technically not challenging. The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed. The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required. This may be only correlation. Before such a claim is published in the scientific literature, it must be supported by convincing evidence. Aspiration to clarify this in future papers does not merit publication of the present paper.	1	2	reprodmed3020015_perova	1
+Thank you for correcting the sentence. The description has been corrected in accordance with your suggestion (page 4, lines 141-142).	2	1	Lines 142-3.“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments. All reported results were observed in at least two independent experiments.	1	2	reprodmed3020015_perova	1
+The error has been corrected (page 8, line 257).	2	1	Line 259. PGRMC1-independe (add t) >>	1	2	reprodmed3020015_perova	1
+The description has been modified to the following (page 8, lines 261-266). “Although we cannot exclude the possible non-specific action of AG-205 on ESCs, the effect of AG-205 treatment and PGRMC1 knockdown promoted in vitro decidualization. These findings suggested that PGRMC1 downregulation may promote ESC decidualization during the secretory phase.	2	1	Lines 263-267: “Because the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence.	1	2	reprodmed3020015_perova	1
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, lines 280-281).	2	1	There are two natural sentences here. However, as described above AG-205 is not a specific inhibitor of PGRMC1. [full stop, new sentence] Further study of PGRMC1 knockout on decidualization is needed.	1	2	reprodmed3020015_perova	1
+As you pointed out, we did not explain the relationship between PGRMC1 stability and post-transcriptional regulation of the protein. We have added the following description (page 9, lines 304-308). ”These reports suggested that PGRMC1 protein levels can be modulated by changes in the rates of transcription, translation, and degradation. Therefore, further studies would be needed to clarify the possible involvement of the post-translational modification and degradation that may regulate PGRMC1 protein stability in ESCs during decidualization.”	2	1	The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation. Sabbir’s results (ref 31) imply that this also holds for PGRMC1. The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned. To be clear, protein levels can be mediated by altered rates of transcription, translation, or degradation. This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues.	1	2	reprodmed3020015_perova	1
+The XGM2016 model is parameterized as a spherical harmonic series expansion resolved to degree and order (d/o) 719, which is the maximum resolution supported by the 15′ terrestrial gravity grid and a satellite-only model GOCO05s. For XGM2016, a significant focus is the optimal combination of the new terrestrial data with the latest satellite gravity information. The combination is based on the rigorous solution of a full normal equation system up to the maximum d/o 719. The calculation of the XGM2019 spheroidal harmonic model coefficients up to d/o 719 consists of a weighted least squares adjustment of GOCO06s with the primary 15′ NGA ground gravity dataset. The XGM2016 and XGM2019 models used the 15′ NGA ground gravity dataset, however, the refined GFMs in this study are obtained by combining the GRACE/GOCE-based GGMs and EGM2008 model, The gravity field information of 5′ terrestrial gravity data in EGM2008 is fully utilized. In addition, to consider the influence of higher frequency gravity field signals caused by topography, the RTM is utilized to further compensate for the omission errors in the refined GGMs. In this study, the combination of the satellite-only GGM with the EGM2008 is based on a pure complementation of the spherical harmonic coefficients at a specific degree, We added comparisons for XGM2016 or XGM2019 models in Table 4, we can find that the refined GFMs outperform XGM2016 and XGM2019 as well, the major improvement of the refined GFMs can be attributed to the GOCE data and topography signals. Please refer to page 14, Line 435-457.	2	1	What is the main methodological difference and advantage of your method? Please comment on that question quantitatifely and qualitatively.	1	2	rs14061437_makarova	1
+"Thank you for your good question and advice. the high-quality GNSS/levelling-based height anomalies are used to check the refined GGMs for obtaining the optimal combination degrees. Thus, the refined GGMs provide better local quasi-geoid results. We have added descriptions and contents in the revised manuscript. The combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree in this paper, which is different from rigorous combination that is done on the basis of the normal equations and co-variance by a least-squares. The purpose of this pure combination can provide better local quasi-geoid results, it is said that it can obtain the characteristics of spatial ""localization"" for quasi-geoid. Combining the satellite-only GFM with EGM2008 by applying some sophisticated weighting approach usually combines the maximum degree and order of the satellite-only GFM and the EGM2008. Because the degree errors of the satellite-only GFM increase with the increase in degree and order, the noise starts to dominate the signals at high degree and order. The noise of satellite-only GFM maybe introduce by sophisticated weighting approach. In addition, the rigorous combination based on the sophisticated weighting approach usually needs the full error variance-covariance matrix of the spherical harmonic coefficients, but, the full error variance-covariance matrix of the spherical harmonic coefficients might generally not be available. Although the obtained results are already quite promising, it can be expected that the refined GFMs provide a guidance for determining the quasi-geoid or the geopotential value of the vertical datum in China. However, the combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree. However, such a procedure might cause a spectral gap between both models. In the next step, the rigorous combination or a smooth transition (such as: using hanning window) will be considered to derived the refined GGMs. Please refer to page 17, Line 544-545."	2	1	Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach? Could you please comment on the effects you introduce by applying such a sharp truncation combination approach.	1	2	rs14061437_makarova	1
+Please refer to page 2, Line 60.	2	1	Page 2, Line 60. Change “,” to “.”	1	2	rs14061437_makarova	1
+Please refer to page 2, Line 63.	2	1	Page 2, Line 62. Change “to use of high-accuracy” to “to use high-accuracy” Thank you.	1	2	rs14061437_makarova	1
+Please refer to page 11, Line 359.	2	1	Page 11, Line 356. Change “not reasonable” to “not a reasonable” Thank you.	1	2	rs14061437_makarova	1
+Thank you for your advice. We have rewritten the mean values. Please refer to page 16, Line 496.	2	1	Page 16, Line 488. Considering the STD you should rather not give the second digit after the comma.	1	2	rs14061437_makarova	1
+Thank you for your comment. The unified topo data and Equation are used to compute RTM quasi-geoid height. We have added some expressions about the using of unified topo data and equations, which makes us easily misunderstand about the calculation process in the text. Please refer to page 12, Line 402.	2	1	Generally it is a good paper and it was interesting to me. However, comparing with the conclusions (I mean, the computed potential value) the paper is a bit too long. I can accept it, because it is a good explanatory text, however some partially offtopic or superfluous parts (e.g. point 2.1.2 (keeping the Table 1 only), the unified topo data in lines 181-191, the „textbook equations” of the gravity of a prism in page 7 and the textbook adjustment equations in page 8). It is rather an editorial question. The paper is good and acceptable without them, too.	1	2	rs14061437_makarova	1
+Thank you. Done. Please refer to page 8, Line 259.	2	1	Line 259 refers to a x as a ’parameter’ in Eq 12, I suggest to use ’vector’ or ’parameter vector’ instead.	1	2	rs14061437_makarova	1
+Thank you for your constructive suggestion. We have moved the discussion part in results to Discussion. The conclusions are recompiled and added according to your suggestions. Please refer to Line 498-550 and Line 551-589.	2	1	As a structural suggestion, considerable part of the Results are rather belong to Discussion (thus making the better balance between these two chapters, in extent and in content, too): I suggest point 3.3 (or maybe also 3.2) to Discussion – as they are discussing the already shown results. Conclusion – it shouldn’t be a summary as should be recompiled. The second part of the 3rd paragraph and the 4th paragraph are enough, completed by the estimated error of the main conclusive potantial value.	1	2	rs14061437_makarova	1
+Please refer to page 2, Line 60.	2	1	Page 2, Line 60. Change “,” to “.”	1	2	rs14061437_perova	1
+Please refer to page 2, Line 63.	2	1	Page 2, Line 62. Change “to use of high-accuracy” to “to use high-accuracy” Thank you.	1	2	rs14061437_perova	1
+Please refer to page 11, Line 359.	2	1	Page 11, Line 356. Change “not reasonable” to “not a reasonable” Thank you.	1	2	rs14061437_perova	1
+Thank you for your advice. We have rewritten the mean values. Please refer to page 16, Line 496.	2	1	Page 16, Line 488. Considering the STD you should rather not give the second digit after the comma.	1	2	rs14061437_perova	1
+The XGM2016 model is parameterized as a spherical harmonic series expansion resolved to degree and order (d/o) 719, which is the maximum resolution supported by the 15′ terrestrial gravity grid and a satellite-only model GOCO05s. For XGM2016, a significant focus is the optimal combination of the new terrestrial data with the latest satellite gravity information. The combination is based on the rigorous solution of a full normal equation system up to the maximum d/o 719. The calculation of the XGM2019 spheroidal harmonic model coefficients up to d/o 719 consists of a weighted least squares adjustment of GOCO06s with the primary 15′ NGA ground gravity dataset. The XGM2016 and XGM2019 models used the 15′ NGA ground gravity dataset, however, the refined GFMs in this study are obtained by combining the GRACE/GOCE-based GGMs and EGM2008 model, The gravity field information of 5′ terrestrial gravity data in EGM2008 is fully utilized. In addition, to consider the influence of higher frequency gravity field signals caused by topography, the RTM is utilized to further compensate for the omission errors in the refined GGMs. In this study, the combination of the satellite-only GGM with the EGM2008 is based on a pure complementation of the spherical harmonic coefficients at a specific degree, We added comparisons for XGM2016 or XGM2019 models in Table 4, we can find that the refined GFMs outperform XGM2016 and XGM2019 as well, the major improvement of the refined GFMs can be attributed to the GOCE data and topography signals. Please refer to page 14, Line 435-457. Table 4. Statistics of the height anomaly differences between GNSS/levelling and six higher-degree GFMs. Unit: (m).	2	1	What is the main difference (except of the terrestrial data over China) to other combined GFMs, such as XGM2016 or XGM2019? What is the main methodological difference and advantage of your method? Please comment on that question quantitatifely and qualitatively.	1	2	rs14061437_perova	1
+"Thank you for your good question and advice. the high-quality GNSS/levelling-based height anomalies are used to check the refined GGMs for obtaining the optimal combination degrees. Thus, the refined GGMs provide better local quasi-geoid results. We have added descriptions and contents in the revised manuscript. The combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree in this paper, which is different from rigorous combination that is done on the basis of the normal equations and co-variance by a least-squares. The purpose of this pure combination can provide better local quasi-geoid results, it is said that it can obtain the characteristics of spatial ""localization"" for quasi-geoid. Combining the satellite-only GFM with EGM2008 by applying some sophisticated weighting approach usually combines the maximum degree and order of the satellite-only GFM and the EGM2008. Because the degree errors of the satellite-only GFM increase with the increase in degree and order, the noise starts to dominate the signals at high degree and order. The noise of satellite-only GFM maybe introduce by sophisticated weighting approach. In addition, the rigorous combination based on the sophisticated weighting approach usually needs the full error variance-covariance matrix of the spherical harmonic coefficients, but, the full error variance-covariance matrix of the spherical harmonic coefficients might generally not be available. Although the obtained results are already quite promising, it can be expected that the refined GFMs provide a guidance for determining the quasi-geoid or the geopotential value of the vertical datum in China. However, the combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree. However, such a procedure might cause a spectral gap between both models. In the next step, the rigorous combination or a smooth transition (such as: using hanning window) will be considered to derived the refined GGMs. Please refer to page 17, Line 544-545."	2	1	3.3) seems quite simple for me and probably not optimal. Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach? Could you please comment on the effects you introduce by applying such a sharp truncation combination approach. Your method for combining the satellite-only GFM with EGM2008 (sec.	1	2	rs14061437_perova	1
+Thank you for your comment. The unified topo data and Equation are used to compute RTM quasi-geoid height. We have added some expressions about the using of unified topo data and equations, which makes us easily misunderstand about the calculation process in the text. Please refer to page 12, Line 402.	2	1	Generally it is a good paper and it was interesting to me. However, comparing with the conclusions (I mean, the computed potential value) the paper is a bit too long. I can accept it, because it is a good explanatory text, however some partially offtopic or superfluous parts (e.g. point 2.1.2 (keeping the Table 1 only), the unified topo data in lines 181-191, the „textbook equations” of the gravity of a prism in page 7 and the textbook adjustment equations in page 8). It is rather an editorial question. The paper is good and acceptable without them, too.	1	2	rs14061437_perova	1
+Thank you. Done. Please refer to page 8, Line 259.	2	1	Line 259 refers to a x as a ’parameter’ in Eq 12, I suggest to use ’vector’ or ’parameter vector’ instead.	1	2	rs14061437_perova	1
+Thank you for your constructive suggestion. We have moved the discussion part in results to Discussion. The conclusions are recompiled and added according to your suggestions. Please refer to Line 498-550 and Line 551-589.	2	1	As a structural suggestion, considerable part of the Results are rather belong to Discussion (thus making the better balance between these two chapters, in extent and in content, too): I suggest point 3.3 (or maybe also 3.2) to Discussion – as they are discussing the already shown results. Conclusion – it shouldn’t be a summary as should be recompiled. The second part of the 3rd paragraph and the 4th paragraph are enough, completed by the estimated error of the main conclusive potantial value.	1	2	rs14061437_perova	1
+I added a table listing the baseline characteristics, including BMI and smoking status, of subjects.	2	1	Please add a table with patients' baseline characteristics, including BMI and smoking status.	1	2	s22072682_makarova	1
+Since this study focuses on AT search, detailed estimation of sample size is not performed. I added in data analysis section.	2	1	Did authors estimate sample size? Please add in data analysis section	1	2	s22072682_makarova	1
+Project identification code is H29-4. I added in 2.1.subjects and 6.patents.	2	1	Please add ethics protocol approval number	1	2	s22072682_makarova	1
+My manuscript is checked by an English proofreader. I will attach an English proofreading certificate.	2	1	I found some minor English errors throughout the paper. Please have a re-check.	1	2	s22072682_makarova	1
+I described the paper's aim.	2	1	I suggest presenting the paper's aim as the last idea in the introduction to make the manuscript easier to read.	1	2	s22072682_makarova	1
+I redrew Figures 1 and 2 and described them as an adaptation of the original publication.	2	1	Given the poor quality of the images (Fig 1 and 2), it might be better to redraw them and describe them as an adaptation of the original publication.	1	2	s22072682_makarova	1
+I described baseline load of the participants in 2.2. Experimental Protocol and Setup.	2	1	The methods do not specify how the baseline load of the participants was determined.	1	2	s22072682_makarova	1
+I revised Figure 3.	2	1	"In figure 3, the indication of the test time appears in the reverse direction. Time ""0"" should be at the start."	1	2	s22072682_makarova	1
+I deleted graphs 5, 8, and 11.	2	1	I recommend deleting graphs 5, 8, and 11 because graphs 7, 10, and 13 show the same information together with lines obtained by analyzing the structural change of the time series.	1	2	s22072682_makarova	1
+We have improved the section to clarify the need to extend this line of research.	2	1	Theoretical issues: although the agency theory and practical function of remuneration committees were described in the manuscript, few efforts were made to give the main theoretical mechanism/logic. Especially since “there does not appear to be any literature focused on examining the association between the constitution of remuneration committees and the reporting of CSR information”, the authors should provide the theoretical link to propose the hypotheses.	1	2	su14020860_makarova	1
+Thank you for your suggestion. From a theoretical point of view, we find various references that explain how CSR index is calculated as the ratio between the aggregation of 140 items focused on environmental, social and economic issues and the total number of items analyzed, which codes as 1 if the firm disclose the CSR information related each item, and 0. Gallego‐Álvarez, I., & Pucheta‐Martínez, M. C. Corporate social responsibility reporting and corporate governance mechanisms: An international outlook from emerging countries. Business Strategy & Development 2020, 3(1), 77–97. Pucheta-Martínez, M. C., & Gallego-Álvarez, I. (2021). The Role of CEO Power on CSR Reporting: The Moderating Effect of Linking CEO Compensation to Shareholder Return. Sustainability, 13(6), 3197. Moreover, we include a Table 1 to justify the items used in the Corporate social responsibility disclosure. Additionally, we have also introduced in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. We had the firm size variable labelled as LTA which is calculated as logarithm of total assets. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	Methodological issues: the dummy of CSR_index is too simple and cannot address the complexity of CSR strategy and CSR disclosure. And so is the independent variable. Many other important control variables should be introduced to the regression, such as, financial performance, the firm size, the firm age, the industry-fixed effects, the country-fixed effects. The results may not be robust without these controls.	1	2	su14020860_makarova	1
+Thanks for your suggestion. I have improved the figure in line with academic norms.	2	1	Other issues: the figure 1 can be re-designed to conform the academic norms.	1	2	su14020860_makarova	1
+Thank you very much for your words. In our modest opinion, the originality of the paper is the analysis of the moderating role played by the proportion of independent directors on boards of directors with the relationship between the constitution of remuneration committees and CSR disclosure. -We have not found previous research focused on this topic using independent directors as moderator variable. We have improved the introduction by adding a paragraph that better motivates the work. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclosure of CSR information due to the scarce existence of previous literature in recent years.	2	1	The article is methodologically OK and well written. However, in general, we see many articles like this in the area of social and environmental reporting. Also, the application of agency theory is so common that reading it over and again in the new studies also makes the readers bored these days. Now it is important that the new researchers try to take the research on this area to a new and interesting level. However, I accept this article only for the moderating variable the authors have tested.	1	2	su14020860_makarova	1
+Thank you very much for this suggestion since, indeed, the introduction was presented with scant motivation. We have corrected this issue and we have included a new paragraph based on remuneration committee and which explains the importance of including this committee for companies. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclsoure of CSR information due to the scarce existence of previous literature in recent years.	2	1	The introduction lacks of motivation. The reader needs to understand why your research question is important.	1	2	su14020860_makarova	1
+In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the theoretical background. In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the section to clarify the need to extend this line of research and have included the following references: Abeysekera, I. (2012). Role of remuneration committee in narrative human capital disclosure. Accounting & Finance, 52, 1-23. Alotaibi, K. O., & Hussainey, K. (2016). Determinants of CSR disclosure quantity and quality: Evidence from non-financial listed firms in Saudi Arabia. International Journal of Disclosure and Governance, 13(4), 364-393. Chhaochharia, V. & Grinstein, Y. (2009). CEO compensation and board structure. The Journal of Finance, 61,(1), 231-261. Kanapathippillai, S., Mihret, D., & Johl, S. (2019). Remuneration committees and attribution disclosures on remuneration decisions: Australian evidence. Journal of business ethics, 158(4), 1063–1082 Suttipun, M. (2021). The influence of board composition on environmental, social and governance (ESG) disclosure of Thai listed companies.	2	1	The theoretical background is weak. I am not convinced on your explanation on why committees would affect CSR.	1	2	su14020860_makarova	1
+Thank you for your suggestion, we have included in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	Your empirical model needs to be reconsidered. Indeed, I believe you have some omitted control variables which may cause an endogeneity issue and drive your findings	1	2	su14020860_makarova	1
+Thank you for this suggestion. In our modest opinion, this paper presents the exhaustive revision and calculation.	2	1	The paper lacks of robustness checks	1	2	su14020860_makarova	1
+Thank you. In the implications paragraph located in the conclusions section, we have added a short text to assess the practical implications.	2	1	The conclusion needs to be rewritten to highlight the practical contributions of the paper	1	2	su14020860_makarova	1
+We have improved the section to clarify the need to extend this line of research.	2	1	although the agency theory and practical function of remuneration committees were described in the manuscript, few efforts were made to give the main theoretical mechanism/logic. Especially since “there does not appear to be any literature focused on examining the association between the constitution of remuneration committees and the reporting of CSR information”, the authors should provide the theoretical link to propose the hypotheses.	1	2	su14020860_perova	1
+Thank you for your suggestion. From a theoretical point of view, we find various references that explain how CSR index is calculated as the ratio between the aggregation of 140 items focused on environmental, social and economic issues and the total number of items analyzed, which codes as 1 if the firm disclose the CSR information related each item, and 0. Gallego‐Álvarez, I., & Pucheta‐Martínez, M. C. Corporate social responsibility reporting and corporate governance mechanisms: An international outlook from emerging countries. Business Strategy & Development 2020, 3(1), 77–97.	2	1	the dummy of CSR_index is too simple and cannot address the complexity of CSR strategy and CSR disclosure. And so is the independent variable. Many other important control variables should be introduced to the regression, such as, financial performance, the firm size, the firm age, the industry-fixed effects, the country-fixed effects. The results may not be robust without these controls.	1	2	su14020860_perova	1
+Thanks for your suggestion. I have improved the figure in line with academic norms.	2	1	the figure 1 can be re-designed to conform the academic norms.	1	2	su14020860_perova	1
+Thank you very much for your words. In our modest opinion, the originality of the paper is the analysis of the moderating role played by the proportion of independent directors on boards of directors with the relationship between the constitution of remuneration committees and CSR disclosure. -We have not found previous research focused on this topic using independent directors as moderator variable. We have improved the introduction by adding a paragraph that better motivates the work. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclosure of CSR information due to the scarce existence of previous literature in recent years.	2	1	The article is methodologically OK and well written. However, in general, we see many articles like this in the area of social and environmental reporting. Also, the application of agency theory is so common that reading it over and again in the new studies also makes the readers bored these days. Now it is important that the new researchers try to take the research on this area to a new and interesting level. However, I accept this article only for the moderating variable the authors have tested.	1	2	su14020860_perova	1
+Authors: Thank you very much for this suggestion since, indeed, the introduction was presented with scant motivation. We have corrected this issue and we have included a new paragraph based on remuneration committee and which explains the importance of including this committee for companies. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclsoure of CSR information due to the scarce existence of previous literature in recent years.	2	1	The paper studies the relationship between remuneration committees and CSR as well as the moderating role of independent directors. While the paper seems interesting I have several concerns that I would like to share with the authors: The introduction lacks of motivation. The reader needs to understand why your research question is important.	1	2	su14020860_perova	1
+In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the theoretical background. In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the section to clarify the need to extend this line of research and have included the following references: Abeysekera, I. (2012). Role of remuneration committee in narrative human capital disclosure. Accounting & Finance, 52, 1-23. Alotaibi, K. O., & Hussainey, K. (2016). Determinants of CSR disclosure quantity and quality: Evidence from non-financial listed firms in Saudi Arabia. International Journal of Disclosure and Governance, 13(4), 364-393. Chhaochharia, V. & Grinstein, Y. (2009). CEO compensation and board structure. The Journal of Finance, 61,(1), 231-261. Kanapathippillai, S., Mihret, D., & Johl, S. (2019). Remuneration committees and attribution disclosures on remuneration decisions: Australian evidence. Journal of business ethics, 158(4), 1063–1082 Suttipun, M. (2021). The influence of board composition on environmental, social and governance (ESG) disclosure of Thai listed companies. International Journal of Disclosure and Governance, 1-12.	2	1	The theoretical background is weak. I am not convinced on your explanation on why committees would affect CSR.	1	2	su14020860_perova	1
+In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Authors: Thank you for your suggestion, we have included in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	Your empirical model needs to be reconsidered. Indeed, I believe you have some omitted control variables which may cause an endogeneity issue and drive your findings	1	2	su14020860_perova	1
+Thank you for this suggestion. In our modest opinion, this paper presents the exhaustive revision and calculation.	2	1	The paper lacks of robustness checks	1	2	su14020860_perova	1
+Thank you. In the implications paragraph located in the conclusions section, we have added a short text to assess the practical implications.	2	1	The conclusion needs to be rewritten to highlight the practical contributions of the paper	1	2	su14020860_perova	1
+Our paper has been substantially revised, and we have addressed the five suggested points for improvements by reviewer 1 (see below for more detail).	2	1	This paper seeks to develop the notion of a Natural Social Contract (NSC) to enable transformative governance based on co-evolution. In a nutshell, to quote, the NSC “involves the fine-tuning of top-down policy and visions with important bottom-up processes (…), and takes place in a hybrid sphere, in which there is room for collaboration between governments, businesses, knowledge institutions and civil society, among others, characterized by Penta-helix models and based on multiple value creation”. Because of this collaborative, polycentric approach, the papers draw on Evolutionary Governance Theory, and develops a perspective of “co-evolutionary steering of interrelated transitions”. The paper’s aspiration is clear and laudable; yet its elaboration is wanting. My recommendation therefore leans towards a strong ‘revise and resubmit’, based on these five points.	1	2	su14052976_makarova	1
+Thanks to a substantial revision our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions.	2	1	First, my main problem is that the paper reads as a compilation of manifestos, rather than one argumentative thread. From begin to end, the paper presents a mix of literature review and substantive argumentation, plus some examples. My recommendation is to set out a clearer agenda, in which the first part delves into the literature to explain the key issue, key concepts and key dilemmas, and a second part presents solutions and directions meeting the key questions and dilemmas. In brief, the paper warrants more focus, edge and structure.	1	2	su14052976_makarova	1
+We have now provided a substantive elaboration on the Natural Social Contract (see section 3), and how contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). More specifically, we have provided more detail on transformative social-ecological innovation (see also the new figure 1 on TSEI), on the mutual gains approach and institutional design principles (see new table 1).	2	1	A second point is the limited elaboration of the social contract, including the natural aspect. A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement. Fig2. moreover, details many aspects. What remains unclear and implicit, however, is the form and operationalisation of such contract. How is the social contract articulated; how is consent manifested? How does one conceive and apply natural ‘design principles’ (669)? This also involves a question of scale: how do agreements at project level chime with societal notions of Social Contracts? How does a project mission contribute to a broader social contract? Much of the joining forces in the hybrid sphere between institutional domains may be primarily instrumental (pooling resources, aligning interests, preventing holdups), below the scope of an (aspired) social contract. What kind of hybrid practices gave shape to the latter? And how then is the aspect of nature taken on board?	1	2	su14052976_makarova	1
+The line-up of the figures (no.	2	1	A third point concerns the link between theory and practice. The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary. Much of this, moreover, is action-oriented, pointing out major carriers and direction of change. Two cases serve as quick illustrations, loosely underwriting some key statements. This presents, in my view, too much of a gap. The paper could do more to translate the general aspects of transformative governance into more specific project aspects, as an evaluative framework for debating the cases.	1	2	su14052976_makarova	1
+We moved up the discussion on governance and steering and elaborated on the generative mechanisms behind a transition to a Natural Social Contract, e.g. visualized by the TSEI framework below, with more attention on three different models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.	2	1	Fourth, the section on co-evolution warrants serious re-thinking. Rather than elaborating the notion and development of NSC, this section largely discusses co-evolutionary approaches, with emphasis on discursive aspects (e.g., metaphors, open concepts and master signifiers). While the section aims to uncover the generative mechanisms behind a ‘transition to a Natural Social Contract’, it lacks focus and thread. In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI). This raises two basic problems. One, the definition and position of NSC in this conceptual medley remains vague. Two, the nature and form of evolutionary variation, selection and dynamics remain unclear. Co-evolution is now associated with all kinds of change-in-tandem, social-sustainable, different institutional actors, discursive items, etc. One wonders whether the notion of transformative change really warrant this evolutionary perspective.	1	2	su14052976_makarova	1
+The question of how the NSC can help to meet and overcome such resistances has been addressed by diving into governance approaches that are capable of dealing with complexity, ambiguity and uncertainty, and with particular attention for multiple value creation and the mutual gains approach (e.g. in section 2). At the same time, we are careful not to present those approaches as simple solutions to those problems.	2	1	Following on the latter, my final point concerns the relation between transformative change and NSC. One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc. On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118). I would suggest the paper can delve deeper into the question of how the NSC can help to meet and overcome such resistances, and to bring forces of change and resistance more in line in the story.	1	2	su14052976_makarova	1
+has been revised	2	1	Source citations are sometimes unclear, as they refer to “	1	2	su14052976_makarova	1
+We have taken out the statement that a NCS is a master signifier and rechecked the English.	4	1	"Language is generally okay. The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the? coevolution"", ""health care"", ""es"" etc....)"	3	2	su14052976_makarova	1
+Below we will respond to the suggestions by reviewer 2	2	1	However, the paper in many parts takes an extreme normative tone that weakens its scientific contribution. It is important that the authors give more emphasis on the scientific evidences of what they defend and anchor their argumentation in such evidences, whether theoretical or empirical.	1	2	su14052976_makarova	1
+Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance. For details see section 2 on page 3-10.	2	1	Make the problematization clearer, exploring what motivates the proposed theoretical study and in which previous studies it is anchored. In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.	1	2	su14052976_makarova	1
+"cf. In the given circumstances, and for the given topic, this seems very appropriate. One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive. Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence. We agree that the ""normative-prescriptive"" tone and use of ""should be"" was excessive. This has been changed where appropriate. At the same time, it is unavoidable in a paper which is partly a manifesto., in the form of a call for the creation of a Natural Social Contract and for transformation-oriented forms of governance. We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use. The need for critical analysis and the presence of limitations to steering are acknowledged. editor’s comment 3 about being normative: First of all, we would like to underline that it is ok to be normative."	2	1	"Remove from the problematization the ""normative-prescriptive"" tone starting from questions and nuisances that justify the study and from gaps in the literature. I am referring to universal and generalist statements with a ""should be"" tone that do not fit well with a scientific article, like the one in lines 51 to 54, among many others in the article."	1	2	su14052976_makarova	1
+The systemic leverage points for transformation offered can be used for achieving change through questions about interdependencies which can be utilised by actor coalitions interested in transformative change through sustainability transitions which are not unduly sectoral but also pay attention to issues of just transition, resilience and the overarching goal of instituting a Natural Social Contract. Achieving this requires and involves a rebalancing of society and new imaginaries (as master signifiers), such as the well-being economy and a Natural Social Contract, as important new orientations.	2	1	Make clearer and more explicit how the article meets the proposed problematic and responds to the gaps found in the debate, including raising the limits of such proposition.	1	2	su14052976_makarova	1
+We fully agree that there is not a single path or model for building public governance that favors sustainability. In this regard, we have stressed, on several occasions, that social contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.	2	1	"Further problematize the discussions/debates that are being mobilized in the article. The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial. It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates. To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy. Only in these aspects it is possible to bring several elements that problematize the discussion and show that there is not a single path or model for building public governance that favors sustainability."	1	2	su14052976_makarova	1
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders. For more information see section 2 on page 3-10. Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance (including Ansell and Gash, 2015), evolutionary steering and adaptive governance.	2	1	In terms of the debate on public governance it would be important to insert the discussion of co-evolutionary governance not only ideally, but also exploring the differences between this theoretical proposition and other forms of governance already discussed in the literature as collaborative governance (Ansell and Gash, 2015), for example. Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.	1	2	su14052976_makarova	1
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4). 2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers. Our line of argumentation has been revised, and examples have been reduced to examples. Thanks to many changes our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions. In addition, the line-up of the figures (no.	2	1	"Empirical cases are very relevant, but are placed as ""mere examples"" without delving into the challenges and difficulties lived in practice. In this sense, it is not clear the relationship between the propositions presented by the authors in section 4 and the empirical or even theoretical evidences presented in the article. I suggest to improve the interface between the propositions presented and what comes before in the text."	1	2	su14052976_makarova	1
+We have improved the focus and structure in ways described below.	4	1	The authors made a visible effort of revision, introducing new sections and discussing better the debates on which their arguments are based. I don't see a problem in a theoretical article to be normative, however some problems in the paper remain and it still need a revision to be published. Specially it needs better clarity in terms of focus and structure. Here are my recommendations:”	3	2	su14052976_makarova	1
+Governance approaches mentioned are now related to Transformative Social-Ecological Innovation (TSEI), which are innovations with explicit transformative goals and attention to mutual gains, and originating from political science and negotiation theory in particular. EGT gives attention to actor/institutions dialectics and power/knowledge dialectics, elements that have taken been into account in the TSEI-framework (figure 1) and related text.	4	1	The text puts in relation 03 new notions that need to be better related by the authors: evolutionary/transformative governance, transformative socioecological innovation and natural social contract. These notions are proposed by the authors and based on their previous work (since they rely on self-citation). Therefore, it seems essential to me to make it clear in each of the sections of the article that deal with these notions from where they start and how they relate to previous debates to support the authors' arguments. It is also essential to make a better connection between these notions, since the relationship between them is not evident per se.”	3	2	su14052976_makarova	1
+Our build-up of argumentation has been improved.	4	1	"In this sense, I recommend to start with the broader discussion of the natural contract (beggining of session 3), ending with the question of how to put in practice this social contract, what challenges (what is the main focus of this text). The line 160 present this question as: ""The big question for steering is: how to initiate and accelerate system changes whose features are not perfectly clear and often times underdeveloped compared to the existing systems? This would make room to bring session 02 that addresses evolutionary/transformative governance to the TSEI.”"	3	2	su14052976_makarova	1
+Changes have been made accordingly. See section with new title: “Transformation pleas and governance approaches for achieving this”. The section now better relates/differentiates evolutionary governance from the other governance approaches mentioned. EGT understands governance as radically evolutionary and is mindful of limitations of any form of governance.	4	1	"The section 02 ""Transformation pleas and approach to achieve this"" can be improved. I recommend reviewing the title referring to the governance debate that is addressed in the section, for example: ""Transformative pleas and approaches of governance to achieve this""...It is also important to better relate/ differentiate the evolutionary governance proposed by authors and the others discussed in the literature (adaptive, reflexive, deliberative, collaborative, etc.). The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution. I suggest that the authors present the debate and the different definitions and then present the notion of GE, relating it to the TSEI.”"	3	2	su14052976_makarova	1
+Practical examples in this section are now more explicit on co-evolutionary steering and multiple value creation insights. Social contract theory helps to recognize duties and rights of citizens and consider issues of rebalancing society, and approaches and mechanisms for achieving this.	4	1	The examples placed in section 03 could compose a section in itself, which sought to relate theory and practical examples (not cases), illustrating the proposed model. It will be nice to put forward the discussion of the conceptual variables of transformative governance proposed.	3	2	su14052976_makarova	1
+This comment is not entirely clear to us. The four proposals for transition policy in section 4 are based on co-evolutionary governance and related discussions in previous sections.	4	1	It remains not clear the relationship between the propositions presented by the authors in section 4 and the discutions in other sections. I suggest to improve the interface between the propositions presented and what comes before in the text, specialy linking with the contributions and limites of the proposal to face this challenges.	3	2	su14052976_makarova	1
+Language and figures have been improved.	4	1	Formal aspects: The text needs a language revision and formatting of the figures to make them more readable. Avoid the excessive use of acronyms and review them throughout the text. I thank the authors and the editor for this opportunity to review and discuss this interesting paper and I hope that my comments help to improve the paper. Best regards.	3	2	su14052976_makarova	1
+Format of figures and tables has been improved where appropriate.	2	1	I'll suggest a format review in the figures and their quotations, as well as the box and the table.	1	2	su14052976_makarova	1
+Thanks to a substantial revision our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions.	2	1	First, my main problem is that the paper reads as a compilation of manifestos, rather than one argumentative thread. From begin to end, the paper presents a mix of literature review and substantive argumentation, plus some examples. My recommendation is to set out a clearer agenda, in which the first part delves into the literature to explain the key issue, key concepts and key dilemmas, and a second part presents solutions and directions meeting the key questions and dilemmas. In brief, the paper warrants more focus, edge and structure.	1	2	su14052976_perova	1
+We have now provided a substantive elaboration on the Natural Social Contract (see section 3), and how contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). More specifically, we have provided more detail on transformative social-ecological innovation (see also the new figure 1 on TSEI), on the mutual gains approach and institutional design principles (see new table 1).	2	1	A second point is the limited elaboration of the social contract, including the natural aspect. A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement. Fig2. moreover, details many aspects. What remains unclear and implicit, however, is the form and operationalisation of such contract. How is the social contract articulated; how is consent manifested? How does one conceive and apply natural ‘design principles’ (669)? This also involves a question of scale: how do agreements at project level chime with societal notions of Social Contracts? How does a project mission contribute to a broader social contract? Much of the joining forces in the hybrid sphere between institutional domains may be primarily instrumental (pooling resources, aligning interests, preventing holdups), below the scope of an (aspired) social contract. What kind of hybrid practices gave shape to the latter? And how then is the aspect of nature taken on board?	1	2	su14052976_perova	1
+The line-up of the figures (no.	2	1	A third point concerns the link between theory and practice. The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary. Much of this, moreover, is action-oriented, pointing out major carriers and direction of change. Two cases serve as quick illustrations, loosely underwriting some key statements. This presents, in my view, too much of a gap. The paper could do more to translate the general aspects of transformative governance into more specific project aspects, as an evaluative framework for debating the cases.	1	2	su14052976_perova	1
+We moved up the discussion on governance and steering and elaborated on the generative mechanisms behind a transition to a Natural Social Contract, e.g. visualized by the TSEI framework below, with more attention on three different models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.	2	1	Fourth, the section on co-evolution warrants serious re-thinking. Rather than elaborating the notion and development of NSC, this section largely discusses co-evolutionary approaches, with emphasis on discursive aspects (e.g., metaphors, open concepts and master signifiers). While the section aims to uncover the generative mechanisms behind a ‘transition to a Natural Social Contract’, it lacks focus and thread. In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI). This raises two basic problems. One, the definition and position of NSC in this conceptual medley remains vague. Two, the nature and form of evolutionary variation, selection and dynamics remain unclear. Co-evolution is now associated with all kinds of change-in-tandem, social-sustainable, different institutional actors, discursive items, etc. One wonders whether the notion of transformative change really warrant this evolutionary perspective.	1	2	su14052976_perova	1
+The question of how the NSC can help to meet and overcome such resistances has been addressed by diving into governance approaches that are capable of dealing with complexity, ambiguity and uncertainty, and with particular attention for multiple value creation and the mutual gains approach (e.g. in section 2). At the same time, we are careful not to present those approaches as simple solutions to those problems.	2	1	Following on the latter, my final point concerns the relation between transformative change and NSC. One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc. On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118). I would suggest the paper can delve deeper into the question of how the NSC can help to meet and overcome such resistances, and to bring forces of change and resistance more in line in the story.	1	2	su14052976_perova	1
+has been revised	2	1	Source citations are sometimes unclear, as they refer to “	1	2	su14052976_perova	1
+We have taken out the statement that a NCS is a master signifier and rechecked the English.	4	1	"This paper has much improved, also because its scope is clearer. I am still puzzled about the selection aspect of EGT, notably the specific roles of metaphors, open concepts and master signifiers play here (as objects and agents of selection) - particularly now NCS has been presented as a master signifier. But that may be for a later argument. Language is generally okay. The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the? coevolution"", ""health care"", ""es"" etc....)”"	3	2	su14052976_perova	1
+Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance. For details see section 2 on page 3-10.	2	1	1) Make the problematization clearer, exploring what motivates the proposed theoretical study and in which previous studies it is anchored. In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.	1	2	su14052976_perova	1
+"cf. In the given circumstances, and for the given topic, this seems very appropriate. One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive. Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence. We agree that the ""normative-prescriptive"" tone and use of ""should be"" was excessive. This has been changed where appropriate. At the same time, it is unavoidable in a paper which is partly a manifesto., in the form of a call for the creation of a Natural Social Contract and for transformation-oriented forms of governance. We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use. The need for critical analysis and the presence of limitations to steering are acknowledged. editor’s comment 3 about being normative: First of all, we would like to underline that it is ok to be normative."	2	1	"2) ""Remove from the problematization the """"normative-prescriptive"""" tone starting from questions and nuisances that justify the study and from gaps in the literature."" I am referring to universal and generalist statements with a ""should be"" tone that do not fit well with a scientific article, like the one in lines 51 to 54, among many others in the article."	1	2	su14052976_perova	1
+The systemic leverage points for transformation offered can be used for achieving change through questions about interdependencies which can be utilised by actor coalitions interested in transformative change through sustainability transitions which are not unduly sectoral but also pay attention to issues of just transition, resilience and the overarching goal of instituting a Natural Social Contract. Achieving this requires and involves a rebalancing of society and new imaginaries (as master signifiers), such as the well-being economy and a Natural Social Contract, as important new orientations.	2	1	3) Make clearer and more explicit how the article meets the proposed problematic and responds to the gaps found in the debate, including raising the limits of such proposition.	1	2	su14052976_perova	1
+We fully agree that there is not a single path or model for building public governance that favors sustainability. In this regard, we have stressed, on several occasions, that social contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.	2	1	"4) Further problematize the discussions/debates that are being mobilized in the article. The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial. It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates. To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy. Only in these aspects it is possible to bring several elements that problematize the discussion and show that there is not a single path or model for building public governance that favors sustainability."	1	2	su14052976_perova	1
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders. For more information see section 2 on page 3-10. Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance (including Ansell and Gash, 2015), evolutionary steering and adaptive governance.	2	1	5) In terms of the debate on public governance it would be important to insert the discussion of co-evolutionary governance not only ideally, but also exploring the differences between this theoretical proposition and other forms of governance already discussed in the literature as collaborative governance (Ansell and Gash, 2015), for example. Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.	1	2	su14052976_perova	1
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4). 2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers. Our line of argumentation has been revised, and examples have been reduced to examples. Thanks to many changes our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions. In addition, the line-up of the figures (no.	2	1	"6) ""Empirical cases are very relevant, but are placed as """"mere examples"""" without delving into the challenges and difficulties lived in practice."" In this sense, it is not clear the relationship between the propositions presented by the authors in section 4 and the empirical or even theoretical evidences presented in the article. I suggest to improve the interface between the propositions presented and what comes before in the text."	1	2	su14052976_perova	1
+Format of figures and tables has been improved where appropriate.	2	1	We want to sincerely thank you for your interest in the Sustannability and for sharing your work with us. I hope this review will encourage your work and the improvement of the text.	1	2	su14052976_perova	1
+We have improved the focus and structure in ways described below.	4	1	1) The text puts in relation 03 new notions that need to be better related by the authors: evolutionary/transformative governance, transformative socioecological innovation and natural social contract. These notions are proposed by the authors and based on their previous work (since they rely on self-citation). Therefore, it seems essential to me to make it clear in each of the sections of the article that deal with these notions from where they start and how they relate to previous debates to support the authors' arguments. It is also essential to make a better connection between these notions, since the relationship between them is not evident per se.”	3	2	su14052976_perova	1
+Governance approaches mentioned are now related to Transformative Social-Ecological Innovation (TSEI), which are innovations with explicit transformative goals and attention to mutual gains, and originating from political science and negotiation theory in particular. EGT gives attention to actor/institutions dialectics and power/knowledge dialectics, elements that have taken been into account in the TSEI-framework (figure 1) and related text.	4	1	"2) In this sense, I recommend to start with the broader discussion of the natural contract (beggining of session 3), ending with the question of how to put in practice this social contract, what challenges (what is the main focus of this text). The line 160 present this question as: ""The big question for steering is: how to initiate and accelerate system changes whose features are not perfectly clear and often times underdeveloped compared to the existing systems? This would make room to bring session 02 that addresses evolutionary/transformative governance to the TSEI.”"	3	2	su14052976_perova	1
+Our build-up of argumentation has been improved.	4	1	"2) The section 02 """"Transformation pleas and approach to achieve this"""" can be improved."" I recommend reviewing the title referring to the governance debate that is addressed in the section, for example: ""Transformative pleas and approaches of governance to achieve this""...It is also important to better relate/ differentiate the evolutionary governance proposed by authors and the others discussed in the literature (adaptive, reflexive, deliberative, collaborative, etc.). The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution. I suggest that the authors present the debate and the different definitions and then present the notion of GE, relating it to the TSEI.”"	3	2	su14052976_perova	1
+Changes have been made accordingly. See section with new title: “Transformation pleas and governance approaches for achieving this”. The section now better relates/differentiates evolutionary governance from the other governance approaches mentioned. EGT understands governance as radically evolutionary and is mindful of limitations of any form of governance.	4	1	3) The examples placed in section 03 could compose a section in itself, which sought to relate theory and practical examples (not cases), illustrating the proposed model. It will be nice to put forward the discussion of the conceptual variables of transformative governance proposed.	3	2	su14052976_perova	1
+Practical examples in this section are now more explicit on co-evolutionary steering and multiple value creation insights. Social contract theory helps to recognize duties and rights of citizens and consider issues of rebalancing society, and approaches and mechanisms for achieving this.	4	1	4) It remains not clear the relationship between the propositions presented by the authors in section 4 and the discutions in other sections. I suggest to improve the interface between the propositions presented and what comes before in the text, specialy linking with the contributions and limites of the proposal to face this challenges.	3	2	su14052976_perova	1
+This comment is not entirely clear to us. The four proposals for transition policy in section 4 are based on co-evolutionary governance and related discussions in previous sections.	4	1	5) Formal aspects: The text needs a language revision and formatting of the figures to make them more readable. Avoid the excessive use of acronyms and review them throughout the text.	3	2	su14052976_perova	1
+Language and figures have been improved.	4	1	Best regards I thank the authors and the editor for this opportunity to review and discuss this interesting paper and I hope that my comments help to improve the paper.	3	2	su14052976_perova	1
+The confusingly written part has been simplified again. In addition, in order to solve the problem of awkward English, we have re-translated the entire sentence.	2	1	The abstract is very confusingly written and needs a lot of editing. This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.	1	2	su14052981_makarova	1
+As your comments, Korea's smart city has technological and normative advances compared to other countries. However, we think that the USA and China are more developed in service than Korea`s (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022). Therefore, this paper intends to cover only general smart city technologies worldwide (Smart city trends: A focus on 5 countries and 15 companies, Cities 2022). We fully agree with the reviewer's comments that they may feel uncomfortable in other countries. After that, we will do more research and expand the case to other countries. Thank you very much for your comments.	2	1	I have some theoretical questions about the applicability of these approaches outside of Korea. For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.) I think many cities and countries will not be comfortable with this approach. I think this needs some exploration.	1	2	su14052981_makarova	1
+4. In this paper, the monitoring of sewage and wastewater for population level assessment of infection mentioned in smart city applications and urban automation was added as a limitation of this study. In the opinion of reviewers, epidemics in cities are not simply spread by one factor. Therefore, various factors should be dealt with, but in this paper, only heat and movement lines, which were mainly dealt with in Korea. The added part is as follows. Conclusion: “… Also, epidemics in cities are not spread simply by one factor. It is important to operate the monitoring system for sewage and wastewater for population level assessment of infection, which is considered to be the most important source of infection. 2 Therefore, they should be considered in future research.	2	1	I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned. I think that is both a big oversight, and an opportunity for critics. I think that piece needs to be included and worked into any assessment of smart cities and pandemics. Additionally, I've included a few other pieces on COVID and smart cities.	1	2	su14052981_makarova	1
+Additionally, smart city services could respond neither timely nor sequentially.	2	1	Confusing or awkward wording in the abstract: “Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense? Just meaning sequential?)	1	2	su14052981_makarova	1
+This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services.	2	1	“a method responding in-time sequential by flexibly combining”	1	2	su14052981_makarova	1
+Second, recommendations on combining or dismissing certain smart city services, as per the needs of coping with COVID-19, are summarized. Third, smart city services must be utilized only for addressing pandemics, as data from the healthcare system consists of personal information. Therefore, smart city services for responding to COVID-19 must be flexible.	2	1	“Second, flexible smart city services are combined and deleted as needed to cope with COVID-19 has 17 been summarized. Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information. Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.” 2)	1	2	su14052981_makarova	1
+The source has been specified. - (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022).	2	1	(pg3) Tables 1 is interesting, but it’s not clear the source of the data. Is this from the literature? (if so, needs sourcing), or a synthesis by the authors? This may be because of the error message in sourcing in the document I received.	1	2	su14052981_makarova	1
+We have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression 'EA' is a unit used only in certain countries.	2	1	(pg4) Table 3 isn’t clear to me. What is “EA”? Not spelled out.	1	2	su14052981_makarova	1
+Thanks for your kind comments.	2	1	(pg5) Table 4 is very useful.	1	2	su14052981_makarova	1
+We also agree with reviewers that it's an ideal situation to use built-in services only when needed and turn them off when not needed. Therefore, in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport(Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data.	2	1	(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen? Are there concrete examples of such systems being built and operated, then just turned off? Who would oppose turning them off? (I’d imagine many stakeholders) 3)	1	2	su14052981_makarova	1
+We rechecked the paper related to “Key missing components/literatures”. We sincerely apologize for repeating basic mistakes. They all have been revised and reflected.	2	1	Monitoring of sewage/wastewater for COVID case estimation Farkas, K., Hillary, L. S., Malham, S. K., McDonald, J. E., & Jones, D. L. (2020). Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19. Current Opinion in Environmental Science & Health, 17, 14-20. Daughton, C. G. (2020). Wastewater surveillance for population-wide Covid-19: the present and future. Science of the Total Environment, 736, 139631. Bogler, A., Packman, A., Furman, A., Gross, A., Kushmaro, A., Ronen, A., ... & Bar-Zeev, E. (2020). Rethinking wastewater risks and monitoring in light of the COVID-19 pandemic. Nature Sustainability, 3(12), 981-990. Jaiswal, R., Agarwal, A., & Negi, R. (2020). Smart solution for reducing the COVID‐19 risk using smart city technology. IET Smart Cities, 2(2), 82-88.	1	2	su14052981_makarova	1
+Thanks for your kind words. The quality of the paper has improved on the reviewer's advice. There are many things that need to be improved in this paper, however we will remember your sincere advices and write more advanced papers later. Thanks again.	4	1	I think the paper, and especially the abstract, are much improved. Both in terms of readability and accessibility, but also in terms of context and framing. I think the paper has the ability to make a contribution to the literature in a way that the previous version may not have been able to.	3	2	su14052981_makarova	1
+- This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.	4	1	I think mentioning the scenario approach in the abstract might be worthwhile, but I don't think it's a huge problem if it isn't mentioned. Either way, I think the paper is in much better shape.	3	2	su14052981_makarova	1
+In order to improve the quality of the thesis, professional translation has been carried out again. We sincerely apologize for the rudimentary mistakes related to some expressions and errors.	2	1	This article advocates for better flexibility (in the sense of adding and removing when necessary) of smart city services and better integration between them. The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model. First, I have a series of concerns regarding the form. There are some phrasing issues and typos throughout the paper.	1	2	su14052981_makarova	1
+References are rechecked and deleted or erroneous references are recited or deleted. We humbly accept the reviewer's opinion that there are many references, and we have deleted references that are deemed unnecessary. We humbly accept the reviewer's comments that there is a quality problem, and We did our best to double-check, revise, and improve the quality of our papers.	2	1	There are referencing issues with sections as well (e.g. line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic. For many references (e.g. [15], [22], [25]) it is impossible to retrieve the source. For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source. All these issues could have been picked up by a simple check. Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.	1	2	su14052981_makarova	1
+We fully agree with the reviewer's comment that integration between services is difficult not only in smart cities but also in other fields. As advised by the reviewer, the situation in Korea was described in Figure 5 (Outbreak of COVID-19 in Korea and the limitations of smart city services) and Table 10 (Limitations of smart city services in each phase). The spread of COVID cannot simply be blocked by smart city services, but by simplifying the current state and proceeding with the thesis, we did not reflect Korea's medical stage with geographic characteristics. We will conduct additional research in the future to fully reflect the reviewer's comments. Thanks for the advice.	2	1	The content of the paper raises major concerns as well. In the introduction, I could not understand the motivation of the paper. The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic. However, I am lacking a concrete example of failure that proves the authors’ point. It is well-known that different services are hard to integrate together (this is not specific to the smart city), but I would have liked the authors to illustrate in light of the pandemic context, and maybe to the context of Korea since their contribution is specific to this geographic context in that it integrates the 12 smart city categories of Korea and the healthcare phases of Korea. The contribution is not presented in the introduction, only the problem is.	1	2	su14052981_makarova	1
+The 'EA' refers to the quantity, but all have been deleted. We sincerely apologize for the basic mistake. We will check in more detail later. We reviewed the literature review and deleted redundant or unnecessary content. Also, we have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression ‘EA’ is a unit used only in certain countries.	2	1	The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper. It is also hard to read, as it contains more tabular content than text. In Table 3, what is an EA?	1	2	su14052981_makarova	1
+Smart city projects are underway all over the world. Also, although smart city services differ by country, the smart city structure has a similar framework to a certain level. Therefore, if we can transform the structure of the smart city, we decided that it would be a great tool to cope with situations such as COVID, even a little.	2	1	Table 4 presents smart city projects of several different countries. It the threats concern COVID only, it does not really make sense, as many of the mentioned services simply do not have this goal (e.g. smart grid). If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature. The authors than write that hese services have a limitation in that cannot predict and prevent transmission of threats such as COVID.	1	2	su14052981_makarova	1
+Like the reviewer's comments, the rest of the table's contents other than those related to pandemic have been deleted.	2	1	Table 5 is useless in my opinion. It is well-known that many standards exist and are not used in every service, which causes integration issues. The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.	1	2	su14052981_makarova	1
+Sections 2.2 and 2.3 have identified errors with the same title. Accordingly, the title has been modified as follows. We apologize once again for repeating basic mistakes. - 2.2 Status of smart city services - 2.3 Status of Pandemic #9.	2	1	Section 2.3 has the same title as Section 2.2.	1	2	su14052981_makarova	1
+We completely agree with the reviewer's sharp points. Accordingly, the contents including Tables 8 and 9 were deleted and reorganized into World Health Organization (WHO), Control and Prevention (CDC) and Korea Centers for Disease Control (KCDC).	2	1	It presents phased plans for pandemics. Several different plans are described, which is useless information for the reader. Indeed, the phases in a Texan county are of no use to understand the authors’ contribution. They could easily break the section down to the essential information that would be the WHO phases and the Korean healthcare phases, and mention that there are global and local plans throughout the world with different phases. Tables 8 and 9 can thus be removed. Table 7 is not very informative, all the phases say that actions from the previous phase should be continued or initiated. I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors. A case study approach restricted to Korea would be more informative and more consistent with the contribution of the paper, which is as I mentioned specific to Korea.	1	2	su14052981_makarova	1
+Thanks for the reviewer's comments. The title of this section 3 has been changed according to the comments of the reviewers.	2	1	Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find. I suggest e.g. “Revised model of smart city service structure”.	1	2	su14052981_makarova	1
+In the case of Korea, rather than finding the infected person in advance, the focus was on identifying and blocking the spread of the infected person. Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately. As a result, the critical point of being identified before an infectious disease spreads was missed. However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard. We agree with the reviewers' opinion that the biggest cause of this COVID-19 transmission is the rapid spread of infected people around them by moving to public places before they show symptoms. Reflecting the reviewer's opinion that it is not clear, the entire sentence has been revised and translated clearly.	2	1	The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV. However, it misses a critical point of COVID. Infected people are in the public space without knowing that they have COVID, since contagion happens before symptoms appears. Once infected people have been identified, they are supposedly in quarantine. Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts? Or do the authors want to check if infected people are outside their home instead of in quarantine? This is not clear to me. Also, and more importantly, the integration and flexibility aspects are not really addressed in the end. The authors do not explain how the services they propose could be integrated with existing ones (e.g. which standards should be used) and how the services can be added or removed. This part is only represented as clouds in the proposed structure model but not discussed further. Therefore, it seems that the contribution breaks down to proposing strict tracking services which feasibility and acceptance by the population is not assessed.	1	2	su14052981_makarova	1
+First, an epidemiological surveyor should decide whether additional collection of personal information is needed. If the answer is yes, the official should seek approval from relevant authorities to get access to the data. For example, as for the location information, separate permission from the National Police Agency is required.” Source: http://www.molit.go.kr/english/USR/BORD0201/m_28286/DTL.jsp?id=eng_mltm_new&mode=view&idx=2931 3) Key missing components/literatures: We fully agree with the reviewers' comments. Therefore, as the limit of this paper, the limit of the ethical aspect was first specified, and in the situation where the service is not needed, the service is deleted in consideration of the ethical aspect in the structure of the smart city. Also in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport (Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data.	2	1	My last concern is the ethical aspects of the proposed services. The authors acknowledge that not addressing these is a limitation of their research. In my opinion, this is more than a limitation, this is essential to discuss given the nature of what the authors propose.	1	2	su14052981_makarova	1
+Also, we really appreciate the accurate comments from the reviewers.	2	1	In summary, my suggestions are to focus the paper on the Korean context, in an in-depth case study approach where authors detail more their contribution, clearly illustrate what the problem is and how their contribution helps solving it.	1	2	su14052981_makarova	1
+Thank you for the reviewer's heartfelt comments. The corrections to the reviewer's concerns are as follows: #2.	4	1	The authors made substantial efforts to take my comments into account, and I commend them for that. However, despite the good improvements, I have some remaining concerns.	3	2	su14052981_makarova	1
+We checked again that errors occurred. After fixing the errors, we rechecked it on others PC. We sincerely apologize for the continued errors. References deleted in the first review were deleted because they were deemed unnecessary. Thanks for the comments #3.	4	1	"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version. Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography. Please feel free to add them back if you feel they should be included."	3	2	su14052981_makarova	1
+We agree with the reviewer's comments. As advised by the reviewer, the content of the scenario was additionally reflected in the abstract. The contents are as follows.	4	1	The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement. I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable. Indeed, if my understanding is correct, they are the factors that cause the limitations of smart city services the authors want to address.	3	2	su14052981_makarova	1
+The description of the characteristics of each city shown in Table 3 was insufficient, as was the reviewer's opinion. Accordingly, the project described in Table 3 was briefly explained and the contents of the article were also revised. - “... As a representative smart city project in Korea, Pilot city, Regulatory sandbox and Smart City Challenge are in progress. To summarize the smart city services used in this project, medical, crime prevention, transportation and environmental services are the most used.” Table 7 also agrees with the reviewer's advice that it does not fit the current content. Accordingly, the contents of Table 7 were deleted and only the contents described in the text were left. Thanks for your comments.	4	1	The authors made some efforts to streamline Section 2, and it is much easier to follow in its current form. Table 3 just shows that medical, crime prevention, transportation, and environmental services are the most frequent, which is already said in the text, but without an explanation of what pilot city, regulatory sandbox, and smart city challenge are, it is difficult to extract anything else from this table. Table 7 does not bring much useful insights as well in its current form. The authors have not responded to my previous comment on this table, I thus redirect them to my review of the previous version. However, some parts still appear unnecessary to me.	3	2	su14052981_makarova	1
+Thank you. As commented by reviewer, the content of Section 3.1 includes the content of Section 2. However, for the 3.2 Expansion of smart city structure that we want to present, I think it is necessary to organize the contents of Section 2 through Section 3.1 and to draw limitations. Of course, it is also correct to understand the structure as the opinions of reviewer. We put a lot of thought into reflecting the opinions. We are sorry; however, we have decided that it is difficult to change the structure at the moment. We ask for your generous understanding of the current structure.	4	1	Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors. I would thus include it in Section 2 instead. Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2. It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.	3	2	su14052981_makarova	1
+Thank you for understanding a little more about our paper. Also, I totally agree with the reviewer's comments about the skepticism of readers. Accordingly, the legal considerations specified in the first review have been added to the text. Its contents are as follows. “… In Korea, disease-related smart city services are legally sanctioned to be used only when necessary to protect personal information.”	4	1	From the additional explanations in the text, I understand that the idea is to track the movements of everyone and then use that information to trace back the contacts of confirmed infected patients. I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations. The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.	3	2	su14052981_makarova	1
+We are sorry for not being clear on the answer to the last review. This paper tried to present a concept for the smart city structure to cope with major situations such as COVID-19. Therefore, it was not possible to present a specific method of adding or removing a smart city system. Currently, smart city services in Korea are trying to implement a structure that adds and deletes when necessary, as in this paper (Reference #73). However, we apologize for not being able to give you a clear answer on this question. After observing the process of implementing the system, we will present a specific method in the thesis we plan to proceed later.	4	1	Finally, and this is my biggest remaining concern, the authors have not responded to my comment regarding the lack of explanations on how the flexible adding and removal of the proposed services would be achieved. This is still unclear to me in the paper. This is a very important point, since it changes what is the contribution of the paper. The current contribution seems to be the proposal of new services to handle the COVID situation, or similar situations in the future. However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure. In other terms, what can we use from the authors' paper to achieve a more flexible smart city service structure?	3	2	su14052981_makarova	1
+Many revisions have been made to this paper according to the reviewer's comments. Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers. In addition, we will use a more advanced paper by reflecting it in the ongoing research. Also, we sincerely appreciate the accurate comments from the reviewers.	4	1	In brief, the authors have made major improvements on the form and good improvements to the content. However, there are still several of my previous comments that were not answered or not answered in a sufficiently convincing way.	3	2	su14052981_makarova	1
+The reference confirmed that the error occurred when the formatting was moved. In addition, errors in the sentences have been reviewed and corrected. We apologize again for the rudimentary mistake.	2	1	"The whole manuscript is full of ""Error! reference source..."" (e.g. line 85)"	1	2	su14052981_makarova	1
+In other words, the structure lacks flexibility	2	1	Line 142: lack of flexibility	1	2	su14052981_makarova	1
+The full terms of the abbreviation are reflected.	2	1	Please always explain abbreviations before use - e.g. line 149	1	2	su14052981_makarova	1
+The incorrect part of the year has been corrected. Thanks for your pointing out #6.	2	1	Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020	1	2	su14052981_makarova	1
+we sincerely apologize. There were many rudimentary mistakes. We corrected the errors by rechecking the paper.	2	1	Lines 352-354: here we have two times Figure 2. Moreover, please bring the text together.	1	2	su14052981_makarova	1
+Missing or erroneous references were rechecked and corrected.	2	1	In this source you can find a whole list of smart city technologies http://www.sinfonia-smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work	1	2	su14052981_makarova	1
+The confusingly written part has been simplified again. In addition, in order to solve the problem of awkward English, we have re-translated the entire sentence.	2	1	The abstract is very confusingly written and needs a lot of editing. This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.	1	2	su14052981_perova	1
+As your comments, Korea's smart city has technological and normative advances compared to other countries. However, we think that the USA and China are more developed in service than Korea`s (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022). Therefore, this paper intends to cover only general smart city technologies worldwide (Smart city trends: A focus on 5 countries and 15 companies, Cities 2022). We fully agree with the reviewer's comments that they may feel uncomfortable in other countries. After that, we will do more research and expand the case to other countries. Thank you very much for your comments.	2	1	I have some theoretical questions about the applicability of these approaches outside of Korea. For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.) I think many cities and countries will not be comfortable with this approach. I think this needs some exploration.	1	2	su14052981_perova	1
+In this paper, the monitoring of sewage and wastewater for population level assessment of infection mentioned in smart city applications and urban automation was added as a limitation of this study. In the opinion of reviewers, epidemics in cities are not simply spread by one factor. Therefore, various factors should be dealt with, but in this paper, only heat and movement lines, which were mainly dealt with in Korea. The added part is as follows.	2	1	I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned. I think that is both a big oversight, and an opportunity for critics. I think that piece needs to be included and worked into any assessment of smart cities and pandemics. Additionally, I've included a few other pieces on COVID and smart cities.	1	2	su14052981_perova	1
+Additionally, smart city services could respond neither timely nor sequentially.	2	1	“Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense? Just meaning sequential?)	1	2	su14052981_perova	1
+This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services.	2	1	“a method responding in-time sequential by flexibly combining” “Second, flexible smart city services are combined and deleted as needed to cope with COVID-19 has 17 been summarized.	1	2	su14052981_perova	1
+Second, recommendations on combining or dismissing certain smart city services, as per the needs of coping with COVID-19, are summarized. Third, smart city services must be utilized only for addressing pandemics, as data from the healthcare system consists of personal information. Therefore, smart city services for responding to COVID-19 must be flexible.	2	1	Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information. Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.” 2) Theoretical/Substantive questions: “Second, flexible smart city services are combined and deleted as needed to cope with COVID 19 has 17 been summarized.	1	2	su14052981_perova	1
+The source has been specified. - (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022).	2	1	(pg3) Tables 1 is interesting, but it’s not clear the source of the data. Is this from the literature? (if so, needs sourcing), or a synthesis by the authors? This may be because of the error message 3 in sourcing in the document I received.	1	2	su14052981_perova	1
+We have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression 'EA' is a unit used only in certain countries.	2	1	(pg4) Table 3 isn’t clear to me. What is “EA”? Not spelled out.	1	2	su14052981_perova	1
+Thanks for your kind comments.	2	1	(pg5) Table 4 is very useful.	1	2	su14052981_perova	1
+We also agree with reviewers that it's an ideal situation to use built-in services only when needed and turn them off when not needed. Therefore, in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport(Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data. First, an epidemiological surveyor should decide whether additional collection of personal information is needed. If the answer is yes, the official should seek approval from relevant authorities to get access to the data. For example, as for the location information, separate permission from the National Police Agency is required.” Source: http://www.molit.go.kr/english/USR/BORD0201/m_28286/DTL.jsp?id=eng_mltm_new&mode=view&idx=2931 3) Key missing components/literatures: We rechecked the paper related to “Key missing components/literatures”.	2	1	(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen? Are there concrete examples of such systems being built and operated, then just turned off? Who would oppose turning them off? (I’d imagine many stakeholders) 3) Key missing components/literatures:	1	2	su14052981_perova	1
+Webb, W., & Toh, C. K. (2020). The smart city and COVID‐19. IET Smart cities, 2(2), 56-57. Inn, T. L. (2020). Smart city technologies take on COVID-19. World Health, 841. Sharifi, A., Khavarian-Garmsir, A. R., & Kummitha, R. K. R. (2021). Contributions of smart city solutions and technologies to resilience against the COVID-19 pandemic: a literature review. Sustainability, 13(14), 8018. Gusikhin, O. (2020). The Impact of COVID-19 Experience on Smart City and Future Mobility. In Smart Cities, Green Technologies, and Intelligent Transport Systems (pp. 308-321). Springer, Cham. Kim, H. M. (2021). Smart cities beyond COVID-19. In Smart Cities for Technological and Social Innovation (pp. 299-308). Academic Press. 4 Inn, T. L. (2020).	2	1	Smart City and COVID	1	2	su14052981_perova	1
+Farkas, K., Hillary, L. S., Malham, S. K., McDonald, J. E., & Jones, D. L. (2020). Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19. Current Opinion in Environmental Science & Health, 17, 14-20. Daughton, C. G. (2020). Wastewater surveillance for population-wide Covid-19: the present and future. Science of the Total Environment, 736, 139631. Bogler, A., Packman, A., Furman, A., Gross, A., Kushmaro, A., Ronen, A., ... & Bar-Zeev, E. (2020). Rethinking wastewater risks and monitoring in light of the COVID-19 pandemic. Nature Sustainability, 3(12), 981-990. Jaiswal, R., Agarwal, A., & Negi, R. (2020). Smart solution for reducing the COVID‐19 risk using smart city technology. IET Smart Cities, 2(2), 82-88.	2	1	Monitoring of sewage/wastewater for COVID case estimation	1	2	su14052981_perova	1
+Thanks for your kind words. The quality of the paper has improved on the reviewer's advice. There are many things that need to be improved in this paper, however we will remember your sincere advices and write more advanced papers later. Thanks again.	4	1	I think the paper, and especially the abstract, are much improved. Both in terms of readability and accessibility, but also in terms of context and framing. I think the paper has the ability to make a contribution to the literature in a way that the previous version may not have been able to.	3	2	su14052981_perova	1
+- This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.	4	1	I think mentioning the scenario approach in the abstract might be worthwhile, but I don't think it's a huge problem if it isn't mentioned. Either way, I think the paper is in much better shape.	3	2	su14052981_perova	1
+In order to improve the quality of the thesis, professional translation has been carried out again. We sincerely apologize for the rudimentary mistakes related to some expressions and errors.	2	1	This article advocates for better flexibility (in the sense of adding and removing when necessary) of smart city services and better integration between them. The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model. First, I have a series of concerns regarding the form. There are some phrasing issues and typos throughout the paper.	1	2	su14052981_perova	1
+References are rechecked and deleted or erroneous references are recited or deleted. We humbly accept the reviewer's opinion that there are many references, and we have deleted references that are deemed unnecessary. We humbly accept the reviewer's comments that there is a quality problem, and We did our best to double-check, revise, and improve the quality of our papers.	2	1	There are referencing issues with sections as well (e.g. line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic. For many references (e.g. [15], [22], [25]) it is impossible to retrieve the source. For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source. All these issues could have been picked up by a simple check. Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.	1	2	su14052981_perova	1
+We fully agree with the reviewer's comment that integration between services is difficult not only in smart cities but also in other fields. As advised by the reviewer, the situation in Korea was described in Figure 5 (Outbreak of COVID-19 in Korea and the limitations of smart city services) and Table 10 (Limitations of smart city services in each phase). The spread of COVID cannot simply be blocked by smart city services, but by simplifying the current state and proceeding with the thesis, we did not reflect Korea's medical stage with geographic characteristics. We will conduct additional research in the future to fully reflect the reviewer's comments. Thanks for the advice.	2	1	The content of the paper raises major concerns as well. In the introduction, I could not understand the motivation of the paper. The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic. However, I am lacking a concrete example of failure that proves the authors’ point. The contribution is not presented in the introduction, only the problem is. It is well-known that different services are hard to integrate together (this is not specific to the smart city), but I would have liked the authors to illustrate in 2 light of the pandemic context, and maybe to the context of Korea since their contribution is specific to this geographic context in that it integrates the 12 smart city categories of Korea and the healthcare phases of Korea.	1	2	su14052981_perova	1
+The 'EA' refers to the quantity, but all have been deleted. We sincerely apologize for the basic mistake. We will check in more detail later. We reviewed the literature review and deleted redundant or unnecessary content. Also, we have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression ‘EA’ is a unit used only in certain countries.	2	1	The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper. It is also hard to read, as it contains more tabular content than text. In Table 3, what is an EA?	1	2	su14052981_perova	1
+Smart city projects are underway all over the world. Also, although smart city services differ by country, the smart city structure has a similar framework to a certain level. Therefore, if we can transform the structure of the smart city, we decided that it would be a great tool to cope with situations such as COVID, even a little.	2	1	Table 4 presents smart city projects of several different countries. It the threats concern COVID only, it does not really make sense, as many of the mentioned services simply do not have this goal (e.g. smart grid). If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature. The authors than write that hese services have a limitation in that cannot predict and prevent transmission of threats such as COVID.	1	2	su14052981_perova	1
+Like the reviewer's comments, the rest of the table's contents other than those related to pandemic have been deleted.	2	1	Table 5 is useless in my opinion. It is well-known that many standards exist and are not used in every service, which causes integration issues. The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.	1	2	su14052981_perova	1
+Sections 2.2 and 2.3 have identified errors with the same title. Accordingly, the title has been modified as follows. We apologize once again for repeating basic mistakes. - 2.2 Status of smart city services - 2.3 Status of Pandemic #9.	2	1	Section 2.3 has the same title as Section 2.2.	1	2	su14052981_perova	1
+We completely agree with the reviewer's sharp points. Accordingly, the contents including Tables 8 and 9 were deleted and reorganized into World Health Organization (WHO), Control and Prevention (CDC) and Korea Centers for Disease Control (KCDC).	2	1	It presents phased plans for pandemics. Several different plans are described, which is useless information for the reader. Indeed, the phases in a Texan county are of no use to understand the authors’ contribution. They could easily break the section down to the essential information that would be the WHO phases and the Korean healthcare phases, and mention that there are global and local plans throughout the world with different phases. Tables 8 and 9 can thus be removed. Table 7 is not very informative, all the phases say that actions from the previous phase should be continued or initiated. I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors. A case study approach restricted to Korea would be more informative and more consistent with the contribution of the paper, which is as I mentioned specific to Korea.	1	2	su14052981_perova	1
+Thanks for the reviewer's comments. The title of this section 3 has been changed according to the comments of the reviewers.	2	1	Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find. I suggest e.g. “Revised model of smart city service structure”.	1	2	su14052981_perova	1
+In the case of Korea, rather than finding the infected person in advance, the focus was on identifying and blocking the spread of the infected person. Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately. As a result, the critical point of being identified before an infectious disease spreads was missed. However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard. We agree with the reviewers' opinion that the biggest cause of this COVID-19 transmission is the rapid spread of infected people around them by moving to public places before they show symptoms. Reflecting the reviewer's opinion that it is not clear, the entire sentence has been revised and translated clearly.	2	1	The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV. However, it misses a critical point of COVID. Infected people are in the public space without knowing that they have COVID, since contagion happens before symptoms appears. Once infected people have been identified, they are supposedly in quarantine. Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts? Or do the authors want to check if infected people are outside their home instead of in quarantine? This is not clear to me. Also, and more importantly, the integration and flexibility aspects are not really addressed in the end. The authors do not explain how the services they propose could be integrated with existing ones (e.g. which standards should be used) and how the services can be added or removed. This part is only represented as clouds in the proposed structure model but not discussed further. Therefore, it seems that the contribution breaks down to proposing strict tracking services which feasibility and acceptance by the population is not assessed.	1	2	su14052981_perova	1
+We fully agree with the reviewers' comments. Therefore, as the limit of this paper, the limit of the ethical aspect was first specified, and in the situation where the service is not needed, the service is deleted in consideration of the ethical aspect in the structure of the smart city. Also in Korea, it is legally required to be used only when necessary.	2	1	My last concern is the ethical aspects of the proposed services. The authors acknowledge that not addressing these is a limitation of their research. In my opinion, this is more than a limitation, this is essential to discuss given the nature of what the authors propose.	1	2	su14052981_perova	1
+Also, we really appreciate the accurate comments from the reviewers.	2	1	In summary, my suggestions are to focus the paper on the Korean context, in an in-depth case study approach where authors detail more their contribution, clearly illustrate what the problem is and how their contribution helps solving it.	1	2	su14052981_perova	1
+Thank you for the reviewer's heartfelt comments. The corrections to the reviewer's concerns are as follows: #2.	4	1	The authors made substantial efforts to take my comments into account, and I commend them for that. However, despite the good improvements, I have some remaining concerns.	3	2	su14052981_perova	1
+We checked again that errors occurred. After fixing the errors, we rechecked it on others PC. We sincerely apologize for the continued errors. References deleted in the first review were deleted because they were deemed unnecessary. Thanks for the comments #3.	4	1	"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version. Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography. Please feel free to add them back if you feel they should be included."	3	2	su14052981_perova	1
+We agree with the reviewer's comments. As advised by the reviewer, the content of the scenario was additionally reflected in the abstract. The contents are as follows.	4	1	The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement. I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable. Indeed, if my understanding is correct, they are the factors that cause the limitations of smart city services the authors want to address.	3	2	su14052981_perova	1
+The description of the characteristics of each city shown in Table 3 was insufficient, as was the reviewer's opinion. Accordingly, the project described in Table 3 was briefly explained and the contents of the article were also revised. - “... As a representative smart city project in Korea, Pilot city, Regulatory sandbox and Smart City Challenge are in progress. To summarize the smart city services used in this project, medical, crime prevention, transportation and environmental services are the most used.” Table 7 also agrees with the reviewer's advice that it does not fit the current content. Accordingly, the contents of Table 7 were deleted and only the contents described in the text were left. Thanks for your comments.	4	1	The authors made some efforts to streamline Section 2, and it is much easier to follow in its current form. Table 7 does not bring much useful insights as well in its current form. The authors have not responded to my previous comment on this table, I thus redirect them to my review of the previous version. However, some parts still appear unnecessary to me. Table 3 just shows that medical, crime prevention, transportation, and environmental services are the most frequent, which is already said in the text, but without an explanation of what pilot city, regulatory sandbox, and smart city 2 challenge are, it is difficult to extract anything else from this table.	3	2	su14052981_perova	1
+Thank you. As commented by reviewer, the content of Section 3.1 includes the content of Section 2. However, for the 3.2 Expansion of smart city structure that we want to present, I think it is necessary to organize the contents of Section 2 through Section 3.1 and to draw limitations. Of course, it is also correct to understand the structure as the opinions of reviewer. We put a lot of thought into reflecting the opinions. We are sorry; however, we have decided that it is difficult to change the structure at the moment. We ask for your generous understanding of the current structure.	4	1	Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors. I would thus include it in Section 2 instead. Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2. It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.	3	2	su14052981_perova	1
+Thank you for understanding a little more about our paper. Also, I totally agree with the reviewer's comments about the skepticism of readers. Accordingly, the legal considerations specified in the first review have been added to the text. Its contents are as follows.	4	1	From the additional explanations in the text, I understand that the idea is to track the movements of everyone and then use that information to trace back the contacts of confirmed infected patients. I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations. The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.	3	2	su14052981_perova	1
+We are sorry for not being clear on the answer to the last review. This paper tried to present a concept for the smart city structure to cope with major situations such as COVID-19. Therefore, it was not possible to present a specific method of adding or removing a smart city system. Currently, smart city services in Korea are trying to implement a structure that adds and deletes when necessary, as in this paper (Reference #73). However, we apologize for not being able to give you a clear answer on this question. After observing the process of implementing the system, we will present a specific method in the thesis we plan to proceed later.	4	1	Finally, and this is my biggest remaining concern, the authors have not responded to my comment regarding the lack of explanations on how the flexible adding and removal of the proposed services would be achieved. This is still unclear to me in the paper. This is a very important point, since it changes what is the contribution of the paper. The current contribution seems to be the proposal of new services to handle the COVID situation, or similar situations in the future. However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure. In other terms, what can we use from the authors' paper to achieve a more flexible smart city service structure?	3	2	su14052981_perova	1
+Many revisions have been made to this paper according to the reviewer's comments. Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers. In addition, we will use a more advanced paper by reflecting it in the ongoing research. Also, we sincerely appreciate the accurate comments from the reviewers.	4	1	In brief, the authors have made major improvements on the form and good improvements to the content. However, there are still several of my previous comments that were not answered or not answered in a sufficiently convincing way.	3	2	su14052981_perova	1
+The reference confirmed that the error occurred when the formatting was moved. In addition, errors in the sentences have been reviewed and corrected. We apologize again for the rudimentary mistake.	2	1	"The whole manuscript is full of ""Error! reference source..."" (e.g. line 85)"	1	2	su14052981_perova	1
+In other words, the structure lacks flexibility	2	1	Line 142: lack of flexibility	1	2	su14052981_perova	1
+The full terms of the abbreviation are reflected.	2	1	Please always explain abbreviations before use - e.g. line 149 Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020	1	2	su14052981_perova	1
+The incorrect part of the year has been corrected. Thanks for your pointing out #6.	2	1	Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020.	1	2	su14052981_perova	1
+we sincerely apologize. There were many rudimentary mistakes. We corrected the errors by rechecking the paper.	2	1	Lines 352-354: here we have two times Figure 2. Moreover, please bring the text together.	1	2	su14052981_perova	1
+Missing or erroneous references were rechecked and corrected.	2	1	In this source you can find a whole list of smart city technologies http://www.sinfonia-smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work	1	2	su14052981_perova	1
+We very much appreciate the contributions.	2	1	Thank you for giving the opportunity to read such an interesting paper.	1	2	su14095590_perova	1
+An analysis of extreme values was carried out, using univariate and multivariate detection methods. The analysis resulted in the elimination of 34 univariate extreme values and 2 multivariate extreme values. All statistical analysis was repeated considering a database of 520 companies.	2	1	For a better and accurate model I recommend the authors to eliminate from the model the extreme value. For example in Fig 2. - workers values greater then 40, community greater than 58, environment 45, customers 46 (the bullets), in Fig 3 - values with global score greater than 130, in Fig. 5 also all the bullets. Then repeat all the statistical analysis.	1	2	su14095590_perova	1
+Thank you for your suggestion. We researched more literature on the subject and adopted a more moderate perspective for assessing normality (|Sk|>2-3, |Ku|>7-10 and |KuMult|>10). As you mention, with the elimination of the extreme values, the skewness and kurtosis values improved significantly, approaching the range -1 to 1.	2	1	In lines 391-395 I would prefer an Sk and Ku between -1 and 1. You have some values that are not included in this interval - maybe because of the outliers from the figures above that I recommend to exclude.	1	2	su14095590_perova	1
+The discussion section has been improved and future research recommendations have been added in the conclusions.	2	1	The paper is interesting; however, it is not clear what the authors are trying to achieve with this study. The authors test a popular assessment tool using confirmatory factor analysis and finds that the model is weak. However, there is no in-depth discussion or recommendations that follow. In fact, the discussion needs to be grounded in previous research findings in regard to CSR/sustainability indices/assessment tools/initiatives. Unfortunately, I am really not sure what the point is here.	1	2	su14095590_perova	1
+The hypothesis was removed since, as mentioned, it doesn't state what we are trying to test in this study.	2	1	Further, the hypothesis does not seem to be written correctly. First, it should be stated prior to the methods section. Second, currently it is descriptive. In other words, it doesn't state what the authors are trying to test in this study.	1	2	su14095590_perova	1
+Major changes have been made to the paper. We hope that this new version may deserve your recommendation for publication.	2	1	Unfortunately, I cannot recommend publication unless major changes are made to the discussion section and possibly recommendations are added for future research and practice.	1	2	su14095590_perova	1
+The manuscript has been revised to improve grammar and english language.	4	1	English language and style are fine/minor spell check required .	3	2	su14095590_perova	1
+The discussion and conclusions sections have been improved (text in blue).	4	1	Are the arguments and discussion of findings coherent, balanced and compelling?	3	2	su14095590_perova	1
+Thanks for the comment. The conclusions section has been improved (text in blue).	4	1	Are the conclusions thoroughly supported by the results presented in the article or referenced in secondary literature?	3	2	su14095590_perova	1
+Thank you. We very much appreciate the comment.	2	1	The paper is well structured and presented in a very meaningful manner. It definitely adds a value to the existed literature.	1	2	su14095590_perova	1
+The argument associated with studies [28] & [29] was added.	2	1	Line 169 page 4, studies [28] & [29] do not reference to any presented argument(s) unless the preceding para.	1	2	su14095590_perova	1
+The words Table and Figure have all been revised to start with a capital letter.	2	1	The word of Table in all text should start with capital letter (e.g. Line 199, L. 223)	1	2	su14095590_perova	1
+Paragraph presented after Table 2 (page 7) has been moved to footnote.	2	1	Paragraph presented after Table 2 (page 7) is a repetitive content to what has been presented in the Table. It could be removed or move to footnote.	1	2	su14095590_perova	1
+• The (***) have been replaced by the p-values (<0.001) in Table - B Impact Assessment - Regression Weights (original model) (current Table 6).	2	1	It is better to show the P-values in Table 3 rather than (***) although a note is indicated in this regard.	1	2	su14095590_perova	1
+Future research recommendations have been added to the conclusions.	2	1	Very good practical recommendations and criticism to the B Corp certification have been presented in the last para. of section (6). No any future research recommendations have been suggested by the respected authors.	1	2	su14095590_perova	1
+The manuscript has been revised. The literature review has been improved, and new references have been added (text in blue).	4	1	Is the content succinctly described and contextualized with respect to previous and present theoretical background and empirical research (if applicable) on the topic? Must be improved Response 4: Thanks for the comment.	3	2	su14095590_perova	1
+The method section has been improved (text in blue).	4	1	Can be improved. Are the research design, questions, hypotheses and methods clearly stated?	3	2	su14095590_perova	1
+We believe that the reviewers' suggestions have been very helpful in improving the manuscript. Thank you for the comment. The manuscript has been entirely revised.	4	1	The text has some improvements, but there are some points that still demand more attention.	3	2	su14095590_perova	1
+Thank you for the suggestion. The title has been clarified in line with the main objective.	2	1	The title demands a review because there isn’t a connection between the first part and the second one. We understand what the authors intend to inform us after reading the paper. Even though this was clear to me, the title doesn’t show the real idea developed in the paper. The objective is a good guide for the title.	1	2	su14095590_perova	1
+The results were improved and the paper's contribution was added.	2	1	The content is well organized, but the results need some to attract the reader. What is the main result of the B assessment with the confirmatory analysis? Besides, the contribution isn’t clear here.	1	2	su14095590_perova	1
+The introduction has been improved, including a better explaination of B Impact Assessment. The use of confirmatory factor analysis has been explained. The aim of the paper was revised in the abstract to be aligned with the one presented in the introduction.	2	1	This section demands more attention because there are some empty spaces to fill. The authors should explain better what is B Impact Assessment. What is its contribution to the structure of a model using Confirmatory Analysis? For me, lines 39 and 40 show the aim of the paper, but the idea is different from the ones presented in the abstract (“The main objective is to understand the B Impact Assessment, verifying its added value in the sustainability process and socio-economic development of the business sector”). The sentence in the Introduction is more complete than the main objective in the Abstract. The ‘areas’ represent a broader goal than the main dimensions of sustainability (Triple Bottom Line).	1	2	su14095590_perova	1
+Lines 70 to 82 have been revised and included in the introduction. Lines 90 to 103 have been revised and decided to be kept in the literature review as they express arguments from important scientific research ( S. Poponi, A. Colantoni, S. R. S. Cividino, and E. M. Mosconi, “The Stakeholders’ perspective within the B corp certification for a circular approach,” Sustain., vol. 11, no. 6, pp. 1–15, 2019, doi: 10.3390/su11061584. / T. De Mendonca and Y. Zhou, “What does targeting ecological sustainability mean for company financial performance?,” Bus. Strateg. Environ., vol. 28, no. 8, pp. 1583–1593, 2019, doi: 10.1002/bse.2334. / C. Putnam Rankin and T. L. Matthews, “Patterns of B Corps Certification: The Role of Institutional, Economic, and Political Resources,” Societies, vol. 10, no. 3, p. 72, 2020, doi: 10.3390/soc10030072.). The literature review was enhanced with scientific information in the fields of sustainability and certification, and, B Lab certification and sustainability.	2	1	The lines from 70 to 82 could be summarized, and cut and paste in the Introduction section. I suggest cutting the text from lines 90 to 103 because it’s quite an advertisement for the B Certificate. The content in this section is exclusively dedicated to the Certification and the Corporation which offers it, there isn’t any scientific information there. The section demands a literature review about the theme, as the relationship between the certificate with adding value, sustainability, and the ‘areas’ governance, workers, community, environment, and customers. What is the idea of the assessment of sustainability, in these areas? This discussion is the basis of the paper, related to the objective, and connected with Figure 1 presented in the next section, Method. There isn’t a literature review in this work. Even 2 though the areas are from B Lab, the authors should study and related them to sustainability and value creation.	1	2	su14095590_perova	1
+Factor analysis involves the examination of interdependence relationship, so it shouldn’t be expressed as a hypothesis that establishes dependence relationships. The economic dimension has been added in the former lines 180 and 181. We clarify that figure 1 is original and was created by the authors. Regarding the data, we confirm that it was taken from the official website, during the period between March and May 2021. The link provided (https://bcorporation.eu/directory) was the one used. In the meantime, the site has been changed and the directory of certified companies can be found in another link. Table 2 has been eliminated. To avoid the repetition of information in Table 2 and the paragraph after, we followed the recommendations of another reviewer by placing the paragraph as a footnote. The reduction in the number of companies was based on the analysis of the database and information collected from B Lab that allowed us to see that there was a change in the measurement model used in B Corp certification during the period January 2017 and March 2021. Thus, to ensure the 3 consistency of the database and the statistical analysis subsequently performed, we reduced the database to a recent period in which the same measurement model was used. Finally, the hypothesis was removed since, as suggested by another reviewer, it doesn't state what we are trying to test in this study.	2	1	In the Abstract, the objective added an economic dimension to the development, but in lines 180 and 181 the impact to be measured includes just environmental and social. It was not clear to me who is the author of Figure 1, the B Lab, or the authors of this paper? There isn’t a source below the Title of Figure 1, though after that seems that B Corporation is the owner of it. The source of data from all companies in the sample isn’t clear, is B Corporation (site) or authors contact all these companies and did the survey? I understood that the data is from the B Corporation website. I accessed the link available in line 191 and there was a message ‘404 page not found, but when I clicked on ‘What is B Corporation’, I found the ‘areas’ and the steps for a company that intends to assess its practices and be certified. The authors should develop more theory than describe a model which has already existed. Table 2 and the paragraph after it is a repetition of Figure 1 and Table 1, could be cut. It’s clever to reduce the number of companies, though it isn’t clear who contacts the companies, the authors or Company B? The hypothesis is interesting when they come from a discussion in the literature review to the field research to test a theory (models, methods, frameworks, etc). Suddenly, appears one in the Method section, in a paper with a limited presentation of a technical model/certificate from an existed company.	1	2	su14095590_perova	1
+"We have replaced the word ""indicator"" with ""aspect"" to eliminate possible confusion with the 5 indicators in the B Impact Assessment Model (Figure 6). The text has been completely revised and the word 'indices' has been used as the standard. The word ‘chapter’ has been replaced by ‘section’. Finally, we are thankful for the good appreciation to the many models studied in our research."	2	1	The sentence from line 137 to line 138 isn’t clear when I saw Figure 6. The authors could use a standard for a plural of index word, ‘indices’ (345) or ‘indexes’ (362). In line 384 the authors should substitute ‘chapter’ for ‘section’. The authors presented and analyzed the variables/areas of certification with many models, it is interesting and is a basis for analysis in the next section.	1	2	su14095590_perova	1
+The topic is “B Corp certification”. The text has been revised and the source has been added (E. Diez-Busto, L. Sanchez-Ruiz, and A. Fernandez-Laviada, “The b corp movement: A systematic literature review,” Sustain., vol. 13, no. 5, pp. 1–17, 2021, doi: 10.3390/su13052508.). Hypothesis 1 has been removed with the justification provided in response 5. Information was added on Cronbach’s Alpha and its impact on the need for B Impact Assessment improvement.	2	1	In the first sentence, what is the topic that the authors mentioned? What source provided that information? The analysis is interesting, but demands more comparisons and explanations for cuts and adds. For me, the answer to Hypothesis 1 isn’t clear. The authors should reflect the presence of that in the text. The Cronbach’s Alpha ‘selected’ just ‘environment’ area of the B Lab model. This is an interesting result, which could be more analyzed with other assessment models presented in the text, to say what the authors wrote at the end of the section.	1	2	su14095590_perova	1
+Thank you for the suggestions. The word was changed to “objective”. The discussion section has been improved in line with the suggestions made in the conclusion.	2	1	Conclusion: I just see one objective, but in the first sentence the authors wrote ‘objectives’. Where are they? The suggestions are interesting, but demand more analysis in the previous section.	1	2	su14095590_perova	1
+We added more coherent theoretical arguments to the discussion based on the most recently published relevant academic papers on the topic (lines 125-133, 153-158, 442-452). The incorporated studies include Yarram and Adapa (2021), Boukattaya and Omri (2021), Shakil (2021), Wang, Wilson & Li, (2021), Cordeiro et al 2020, Birindelli, et al. (2019), and Nuber & Velte (2021). In total, the list of references increased from 21 to 36. The details of these studies and the logic behind their inclusion are explained in the text (pages 3 and 4). For the quantitative papers, our guiding line is the inclusion of papers proposing a reliable strategy to address the problem of endogeneity characterizing virtually all studies dealing with gender diversity. In a nutshell, gender diversity and firms’ performances (be they financial, ESG or environmental) have reverse causality. More gender-diverse boards are found to improve firms’ performances, and at the same time, better performing firms are more attractive to females. Any study incorporating these two variables would suffer from endogeneity. Therefore, inference and results are biased. Finding an instrument for gender diversity becomes a concern of first importance. The task is challenging as all potential candidates for being a suitable instrument should be already part of the regression equation. So far, the literature has not found an instrument unanimously agreed upon. This is the reason for which our selection of papers is rather limited. We focused on those papers with a choice of instrumental variables that are well explained and papers that ‘’go around’’ the problem by making use of a method that does not rely on external instruments. This includes papers using difference-GMM or system-GMM estimators. Their principal advantage is the use of internal instruments instead of external ones. Indeed, the instruments chosen are difference and the level of the covariates themselves. There are only a few of these studies available, including ours. This again explains the relatively restricted number of references we are considering.	2	1	1- Need to update the literature review with recent studies	1	2	su14127346_makarova	1
+Initially, we opted for integrating the theoretical background disseminated throughout the text to connect to the findings of related papers and to avoid redundancy and repetition. Changes that we incorporated in response to this comment improve the overall theoretical framework of the paper, including concepts, definitions and links to relevant literature. We also articulated theoretical assumptions and added a comprehensive discussion of the observed phenomena. Specifically, we added critical mass theory discussion in the literature review section (lines 125-132). We added interpretation of corresponding results in the findings and discussions section (lines 445-452) and the conclusion section (lines 482-503). These changes in addition to the existing discussions of theories and concepts significantly improve the theoretical background of the paper.	2	1	2- Please add a theoretical background section	1	2	su14127346_makarova	1
+We also emphasize the policy implications of these findings (lines 453-462, 497-503). To better reflect the policy implications of the study, we incorporate changes that are diffuse through the text. In particular, we argue that promoting gender diversity cannot deliver its full potential, and may even lead to adverse results unless it is accompanied by measures that mitigate political and economic uncertainty. These measures would create an enabling environment both for households and entrepreneurs. Such measures should increase agents’ confidence, dump their willingness to invest and expand the activity by reducing their aversion to risk. The accompanying measures should primarily target building and strengthening institutions to unlock this potential. Institutions are a defence against uncertainty. They contribute to anchoring economic agents’ expectations and stabilize the economy. We also elaborate on the finding of the critical mass theory. This result suggests that a very effective way to improve environmental quality and mitigate the adverse effects of economic activities is to encourage gender diversity in corporate boards in countries with low uncertainty where we find a higher impact on firms’ environmental performances. In these countries, the margin for progress is large and the effect of improving gender diversity in corporate boards materializes more easily and at lower costs, as these countries are far from their efficiency frontiers (lines 445-461 and 482-503).	2	1	3- What are the policy implications of your study	1	2	su14127346_makarova	1
+We addressed your suggestion to improve arguments and discussion of findings by revising existing and adding new paragraphs better explaining regression coefficients that demonstrate how political and economic uncertainty shapes the relationship between corporate board gender diversity and firms’ environmental performance. (Lines 379-389). Furthermore, we added supporting arguments disseminated across the text (particularly in lines 348-361, lines 379-389, etc). These relate our finding to theory. We focus here on the attitude towards risk and uncertainties. Women on corporate boards tend to be more risk-averse and lean more toward conservatism in uncertain environments. Therefore, the more females there are on corporate boards, the higher their weight in the board decisions would be, and by the same token, the higher the effect of uncertainty on firms’ decisions. Moreover, we also relate our findings to the critical mass theory, suggesting that the impact of gender diversity on the environmental performances of firms materializes only when a critical mass weight of females is realized in corporate boards (Konrad et al. 2008, Torchia et al. 2011, Schwartz-Ziv, 2017- page 12, lines 445-451).	2	1	4- Add discussion of results section and relate your results with theories.	1	2	su14127346_makarova	1
+Absolutely. Changed accordingly (line 160).	2	1	"""To the best of our knowledge, this is the only study that addresses the role of economic and political uncertainty as a moderator in a model that accounts for the dynamic patterns of firm environmental performance"" (line 128). --> I believe the authors intended ""moderating role."""	1	2	su14127346_makarova	1
+We answered this comment along different dimensions. We revise and elaborate on the explanation and interpretation of the results. We provide further explanations on the ways political and economic uncertainty shapes the relationship between corporate board gender diversity and firms’ environmental performance. We also emphasize the policy implications of these findings (lines 453-461, 487-496). In addition, we add more theoretical arguments to the discussion based on the most recently published academic papers on the topic. (Lines 421-452) “	2	1	The authors could go into greater detail to explain the logic behind this relationship. As it stands, the paper offers little insight into the reasoning behind their hypothesis.” We answered this comment along different dimensions.	1	2	su14127346_makarova	1
+The related theory posits that females’ attitude when facing uncertainty is different from males. They tend to exhibit a strong inclination to wait until the uncertainty is resolved before making major business decisions. To address this comment, we added supporting arguments disseminated across the text (particularly in lines 348-361, lines 421-428, etc). These explain the finding you are referring to by attitude towards risk and uncertainties. Women on corporate boards tend to be more risk-averse and lean more toward conservatism in an uncertain environment. Therefore, the more females there are on corporate boards, the higher would be their weight in the board decisions, and by the same token, the higher would be the effect of uncertainty on firms’ decisions.	2	1	It would be useful if the authors provided some speculative thoughts on the finding that the slope of the GenDiv variable is negative in highly uncertain countries on the firm's environmental performance (Figure 3 (c) red line).	1	2	su14127346_makarova	1
+The literature on the relationship between gender diversity and environmental performances of firms that addresses dynamic endogeneity is rather limited. We mention Gaio and Gonçalves (2022), Lu and Herremans (2019), Kassini et al (2016), Silla, et al (2016) that opt for a dynamic modelling using a System-GMM estimator. This estimator is particularly suitable to panels with large number of individual and few time periods, which the case for our paper. Moreover, it accommodates situations where the independent variable changes slowly (Antoniou et al., 2008). To keep a good flow in the text, we only mention these papers in the introduction section. We elaborate further on them in the literature review section. We describe their main purpose and explain how our analysis departs from them.	2	1	1- In lines 28 and onwards, you argue a lack of literature looking at dynamic (GMM) analysis of the role of female on CSR and ESG issues, but fail to mention some examples. Please provide some, and position your work in that (brief) strand of literature. One example you should consider is Gaio, C., & Gonçalves, T. C. (2022). Gender diversity on the board and firms’ corporate social responsibility. International Journal of Financial Studies, 10(1), 15.	1	2	su14127346_makarova	1
+Thanks for the Suggestion. We highlighted the research hypothesis by moving it the introduction. We also added another hypothesis. The first relates to the very existence of a relationship between gender diversity on corporate boards and firms’ performances. The second, deals with the intensity and shape of this relationship. We believe, this adds a deal of clarity to the text and purpose of the paper.	2	1	2- Please revise if it makes sense to present your research hypothesis in the introduction.	1	2	su14127346_makarova	1
+We reviewed all the text and changed wording in different parts of the text.	2	1	"3- Please reconsider the use of adjectives such as ""complicated"" (line 79), interestingly (line 88) and so on."	1	2	su14127346_makarova	1
+We initially meant, among all these papers, there is one that… We rephrased this sentence and reviewed all the text. We edited sentences when needed.	2	1	"4- What do you mean by ""in the crowd"" (line 141). Please revise carefully english for clarity."	1	2	su14127346_makarova	1
+In our initial submission, we used winsorized data for all our econometric analysis. However, we, inadvertently, displayed summary statistics (Table 2) for raw data, i.e., before winsorizing. We should have reported only winsorized summary statistics. This is precisely what we do in this revised version of the paper. The reason for winsorizing data (beyond its advantage of having regular statistical distribution) was that we initially have detected outliers. Further investigation revealed that one company (RIC: OBE.TO; CompName: Obsidian Energy Ltd) is responsible for these outliers. The company had presumably reported wrong values in FY-12, where PP&E was higher than Total Assets (see table below from Refinitiv Eikon). We have reported this case to Refinitiv (Case number: 11297339) and our doubts were confirmed (See their response below). Refinitiv Eikon replaced old value of PP&E (27834189009) with a correct value (10788172656). In order to avoid similar mistakes, we opted for winsorizing data and avoid outliers. Similarly, only one company had negative Tobins’ Q in 2017 (RIC: ELR.TO ; company name: Eastern Platinum Ltd), which was a reporting typo as well. However, our analysis is not affected by these outliers which are removed after winsorizing all the variables. In this revised version of the paper, we replace the table of summary statistics with the one based on winsorized variables.	2	1	5- Please revise your data: your max tangibility is over 2 (which means that Net PP&E, a subset of Assets, is 2 times those same Assets - this should be a typo. Similar problems might exist for a negative tobin's Q.	1	2	su14127346_makarova	1
+The sentence “The global average of corporate boards’ gender diversity for all the sectors during 2008-2018 was about 13.5%” is correct as it is calculated based on all industries (5120 firms in total). However, since we are focusing on “Mining, Quarrying, and Oil and Gas Extraction Industry”, it makes more sense indeed to report the value for that particular sector. Therefore, we update the text as follows: “The average of corporate boards’ gender diversity for Mining, Quarrying, and Oil and Gas Extraction Industry during 2008-2018 was about 9.6%”.	2	1	"6 - In line 192 you mention ""all the sectors"". But you are working one sector only, aren't you?"	1	2	su14127346_makarova	1
+This is an explanation of the following sentence in our article: “To avoid bias and inconsistency that might be caused by instruments proliferation, we collapse the instruments matrix.” We are using longer lags of the dependent variable as additional instruments. Intuitively, there is a trade-off, on one hand, using longer lags gives more information and improves efficiency, but on the other hand sample gets smaller (as it is the case in standard 2SLS models). GMM is a way around this trade-off between lag length and sample length. However, implementation of System-GMM is not exempt from weaknesses. With the instruments count being quadratic in the time dimension, the variance matrix of the moments can be very large. Estimating its elements may be problematic in particular in a finite sample lacking the adequate amount of information. This finite sample bias can weaken Hansen J-test and overstate its p-value and therefore generate suspicious high pass rate of instruments validity. The problem is known as instruments proliferation (Roodman 2009). To avoid bias and inconsistency due to instruments proliferation, we limit the number of instruments used and collapse the instrument matrix which prevents the model from being over-fitted.	2	1	7- Explain your sentences in lines 242-244. What did you do specifically?	1	2	su14127346_makarova	1
+We revised the concerned section and rewrote the text in order to improve on clarity. The revised text reads as follows: Greater gender diversity for a firm located in a country with high political and economic uncertainty would lower the firm’s environmental performance (0.616-0.853=-0.237).	2	1	"8- Lines 330 and onwards are presenting conclusions that are not warranted by the results and the english is a bit strange (what do you measnby ""households' anxiety""?). Similar problems stem from the following paragraphs discussing the results."	1	2	su14127346_makarova	1
+the inoculation points were labeled with the red arrows in Figure 1 as suggested.	2	1	I would like to suggest authors label inoculation points in Figure 1, which could give readers a clear view of three different inoculation methods.	1	2	toxins14060409_makarova	1
+Sorry for carelessness. We corrected it as suggested. (Pag.4 Line 157) Point 3: Line 143: “PPSD-premature spike death” should be “PPSD-proportion of premature spike death”.	2	1	Line 142: “proportion of PPSD” should be “proportion of PSD or PPSD”.	1	2	toxins14060409_makarova	1
+We revised the expression in the footnote of Table 1. (Pag.4 Line 158) Point 4: Lines 168 - 169: Should it be that DON content of most lines without Fhb1 exceeded 1000 µg/kg under UBFI?	2	1	Line 143: “PPSD-premature spike death” should be “PPSD-proportion of premature spike death”.	1	2	toxins14060409_makarova	1
+we revised the sentence as “DON content of most lines without Fhb1 exceeded 1000 μg∙kg-1 under UBFI method (Figure 2), with the highest reaching 4224.38 μg∙kg-1 (Table S1)”. (Pag.5 Line 184-186) Point 5: Lines 411 & 413: “PPSB” should be “PPSD”.	2	1	Lines 168 - 169: Should it be that DON content of most lines without Fhb1 exceeded 1000 µg/kg under UBFI? Where is 4224.38 µg/kg in Table 1? Do you mean in Table S1?	1	2	toxins14060409_makarova	1
+We corrected it. (Pag.11 Line 441 & 443) Point 6: Table S1: “PPSD - premature spike death” in the footnote should be “PPSD - proportion of premature spike death”.	2	1	Lines 411 & 413: “PPSB” should be “PPSD”.	1	2	toxins14060409_makarova	1
+We revised the expression in the footnote of Table S1.	2	1	Table S1: “PPSD - premature spike death” in the footnote should be “PPSD - proportion of premature spike death”.	1	2	toxins14060409_makarova	1
+Thanks. We revised the annotations in Table S1.	2	1	Fhb1- and Fhb1+ don’t appear in Table S1, so they can be removed from the footnote.	1	2	toxins14060409_makarova	1
+We agree with the reviewer’s comments and make Figure S4 as Figure 4 in this revised version.	2	1	I would suggest to avoid citing supplementary figures at the beginning of the chapter. See for example chapter 2.4. Given that 2.4 chapter discusses data of supplementary figure 4 I would suggest to make figure S4 as Figure 4 of the manuscript.	1	2	toxins14060409_makarova	1
+We marked the inoculation sites with red arrows in Figure 1.	2	1	To futher clarify the infection methods a supplementary figure detailing graphically the modes of inoculation with figures would make the paper extremely useful for the community	1	2	toxins14060409_makarova	1
+We added more details about the strain and cited the following paper. (Pag.10 Line 402-403) Zhang, Y.; Li, A.; Zhu, S.; Li, L.; He, X.; Sun, Z.; Li, T. Basal Rachis Internode Injection (BRII): A novel inoculation method to evaluate wheat resistance to Fusarium head blight. Phytopathology, 2021, 111, 1670-1674, doi:10.1094/PHYTO-11-20-0488-R. Point 4: Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication) Response 4: Thanks!	2	1	Identity of the strain used for infection should be confirmed: a multilocus species characterisation is needed to confirm the species of the strain or a reference to a publication where the strain was described and appropriately characterised.	1	2	toxins14060409_makarova	1
+Thanks! We uploaded the raw mycotoxin data as suggested.	2	1	Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication)	1	2	toxins14060409_makarova	1
+Following the suggestion of this reviewer, we have removed the movie in the revised version.	2	1	The largest concern is regarding the colocalization of IFITM3 and Rab11A. It seems (as the authors state) that these represent abortive events (Figure 7). Therefore the inclusion of the movie examining the colocalization of Rab11A GFP, IFITM3-SNAP, and labeled virion seems completely unnecessary. In addition, the movie does not provide any details on cellular features and a viewer is left wondering where the foci are coming from and going. This reviewer suggests removing the movie or alternatively including labels on the movie of cellular features and including a scale bar.	1	2	v11060548_makarova	1
+We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.	2	1	b. The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?	1	2	v11060548_makarova	1
+The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.”	2	1	c. Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.	1	2	v11060548_makarova	1
+We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.	2	1	d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.	1	2	v11060548_makarova	1
+We have made the suggested changes.	2	1	Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.	1	2	v11060548_makarova	1
+We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.	2	1	Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.	1	2	v11060548_makarova	1
+To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.	2	1	Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.	1	2	v11060548_makarova	1
+If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.	2	1	Figure 4: Uninfected control should be shown in the main text.	1	2	v11060548_makarova	1
+We have made the requested changes.	2	1	Figure 4: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_makarova	1
+We have made the requested changes to figure 5A/C and Figure 7.	2	1	Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_makarova	1
+We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).	2	1	Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.	1	2	v11060548_makarova	1
+Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)	2	1	Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.	1	2	v11060548_makarova	1
+All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.	2	1	Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.	1	2	v11060548_makarova	1
+Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.	2	1	Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.	1	2	v11060548_makarova	1
+"We thank the reviewer for pointing out the inaccuracy in the references. The statement ""...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection"" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3."	2	1	Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.	1	2	v11060548_makarova	1
+The supplemental figures only include experiments that reproduce previously published work or are not absolutely necessary to support the major and most novel findings of the paper. We therefore suggest to keep these data in the supplement.	2	1	Supplemental figures should be included in the main text.	1	2	v11060548_perova	1
+We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.	2	1	The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?	1	2	v11060548_perova	1
+The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.” d.	2	1	Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.	1	2	v11060548_perova	1
+We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.	2	1	Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.	1	2	v11060548_perova	1
+We have made the suggested changes.	2	1	Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.	1	2	v11060548_perova	1
+We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.	2	1	Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.	1	2	v11060548_perova	1
+To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.	2	1	Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.	1	2	v11060548_perova	1
+Figure 2B shows that the amount of IFITM3 is not significantly altered up to 6 h p.i. IFITM3 protein increased significantly after longer infection times such as 24 h p.i. (Fig. S2B), as was previously described by others.	2	1	Figure 3A: Why is there so much IFITM3 present in the uninfected cells? This does not appear to increase post infection as was shown previously.	1	2	v11060548_perova	1
+If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.	2	1	Figure 4: Uninfected control should be shown in the main text.	1	2	v11060548_perova	1
+We have made the requested changes.	2	1	Figure 4: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_perova	1
+We have made the requested changes to figure 5A/C and Figure 7.	2	1	Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_perova	1
+We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).	2	1	Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.	1	2	v11060548_perova	1
+Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)	2	1	Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.	1	2	v11060548_perova	1
+All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.	2	1	Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.	1	2	v11060548_perova	1
+Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.	2	1	Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.	1	2	v11060548_perova	1
+"We thank the reviewer for pointing out the inaccuracy in the references. The statement ""...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection"" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3."	2	1	Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.	1	2	v11060548_perova	1
+The innovation of this research has been clarified in the abstract, the introduction, and the conclusion.	2	1	The innovation of this research needs to be better explored.	1	2	w14030367_perova	1
+Three references have been cited. Please see lines 40-41 in the revised manuscript.	2	1	Cite the following references: Panagopoulos, A. (2021). Energetic, economic and environmental assessment of zero liquid discharge (ZLD) brackish water and seawater desalination systems. Energy Conversion and Management, 235. Techno-economic assessment of Minimal Liquid Discharge (MLD) treatment systems for saline wastewater (brine) management and treatment. Process Safety and Environmental Protection, 146, pp. 656-669. Study and evaluation of the characteristics of saline wastewater (brine) produced by desalination and industrial plants. Environmental Science and Pollution Research, 1-14. Lines 33-38: You should mention that discharge of PPCPs degrades water quality and thus it cannot be directly used for potable water and industrial applications.	1	2	w14030367_perova	1
+Thanks very much for the reviewer. The conclusion has been revised according to the reviewer’s suggestion. The added paragraph in the revised manuscript is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conventional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.	2	1	Conclusion: Discuss the applicability of your findings/results and future study in this field.	1	2	w14030367_perova	1
+Thanks for the reviewer’s suggestion. The conclusion has been revised and integrated into two paragraphs. The first paragraph mainly includes important findings, and the second paragraph mainly includes the outlook for the future in this field.	2	1	Conclusion: Make it as one or two paragraphs.	1	2	w14030367_perova	1
+The salinity concentrations in the samples have been added to Table 1 in the revised manuscript.	2	1	Table 1: What is the salinity (in mg/L) of the samples ?	1	2	w14030367_perova	1
+We agree with the suggestion and comments of the reviewer. All comments have been revised one by one, and the modifications are highlighted in yellow in the marked revised manuscript.	2	1	However, there are some mistakes in the writing of the paper.	1	2	w14030367_perova	1
+Thanks for the reviewer’s helpful suggestion. The objective statement has been added to the abstract to support the seriousness of the problem, and the revised sentences are as follows: Biological activated carbon (BAC) biofilter coupling ultrafiltration (UF) is a promising process for the treatment of river water contaminated by pharmaceutical and personal care products (PPCPs). However, the pilot-scale study should be conducted to reveal the long-term removal performance and the respective contributions of BAC and UF.	2	1	The title seems good, but the abstract seems to be fine. Please add one problem statement line in abstract to justify this sentence ``the long-term pilot-scale study is urged to be investigated.``.	1	2	w14030367_perova	1
+We agree with the suggestion of this reviewer. The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances. However, the mechanisms of the two-stage biofilm, such as bacterial and metazoan communities, membrane fouling and dissolved oxygen transfer, should be further investigated to enhance the removal efficiency and stability of this system. The research gap has been added in the Conclusion and is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conven-tional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.	2	1	Research gap should be delivered on more clear way with directed necessity for the future research work.	1	2	w14030367_perova	1
+Thanks for the reviewer’s comment. The introduction has been revised carefully, including using up-to-date references. For example, the outdated references have been removed. Meanwhile, the articles published within the last two years were added, such as (the Reference2 Yu et al., 2020) and (the Reference21 Tang et al., 2018).	2	1	Introduction section must be written on more quality way, i.e., more up-to-date references addressed. Please target the specific gap such as 2015-2021 etc.	1	2	w14030367_perova	1
+The reference has been cited. Please see line 38 in the revised manuscript.	2	1	Page 1 Line 38. Please cite this reference with existing reference 3….Role of nanotechnology for design and development of cosmeceutical: application in makeup and skin care.	1	2	w14030367_perova	1
+The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water. The above coupling process makes up for the defects concerning respective operations of the BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the different potential of biofilm growing. As far as we investigated, the lack of enough attention to long-term pilot study exists. Therefore, in this study, a BAC-UF system was carried out for several months with pilot scale to access the long-term removal performances and the respective contributions of BAC and UF. The relevant content has been supplemented and revised in the abstract to highlight the innovative points.	2	1	The novelty of the work must be clearly addressed and discussed, compare previous research with existing research findings and highlight novelty.	1	2	w14030367_perova	1
+Biological activated carbon (BAC) combined the adsorption and biologic degradation consuming low power energy and chemicals without concern of DBPs production as well as no frequent updates for activated carbon media. The BAC biofilter can remove the PPCPs pollutants, and then the followed UF can reject microorganisms and particles flowing out from the biofilter to ensure the quality of drinking water. Thus, the BAC-UF process makes up for the defects of the respective operations of BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the potential of biofilm growing differently. As far as we investigated, the lack of enough attention to long-term pilot study is present. Therefore, in this study, a BAC-UF system was carried out for several months with a pilot scale.	2	1	What is the main challenge? Why author choose this material? Please highlight in the introduction part.	1	2	w14030367_perova	1
+The two references have been cited. Please see the lines 58.	2	1	Page 2 Line 55 need a reference. Please consider these at end of this sentence……The oxidation method exhibited a fast reaction speed and high removal efficiency…(i) Role of nanomaterials in the treatment of wastewater: A review (ii) Advances and challenges in developing efficient graphene oxide-based ZnO photocatalysts for dye photo-oxidation.	1	2	w14030367_perova	1
+The introduction has been improved for better reading.	2	1	The main objective of the work must be written on the more clear and more concise way at the end of introduction section.	1	2	w14030367_perova	1
+Two PPCP mistakes have been revised. The abbreviation of chemical oxygen demand in previous version has been corrected to CODMn. The unnecessary abbreviations have been also corrected in the abstract.	2	1	Please check the abbreviations of words throughout the article. All should be consistent.	1	2	w14030367_perova	1
+The added paragraph is as follows: Potassium permanganate, H2SO4, potassium sodium tartrate tetrahydrate, Nessler’s reagent and NaOH were purchased from a commercial company and certified as AR purity (Guangzhou Chemical Reagent Factory, Guangzhou, China), while PPCP standards were provided by three companies. Sulfamethoxazole (SMX), Sulfadoxine (SD), Sulfachloropyridazine (SCP), Sulfamethoxypyridazine (SMP), Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Erythromycin (EM), Anhydroerythromycin (EA), Roxithromycin (ROX), Penicillin-G (PEN G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacine (IND), Metoprolol (METO), Propranolol (), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), and Sulpiride (SP) were obtained from the Dr. Ehrenstorfer Company in Germany. Furthermore, Sulfadimidine (SM2), Sulfadiazine (SDZ), Sulfapyridine (SP), Sulfamonomethoxine (SMM), Tetracycline (TC), Ofloxacin (OFL), Amoxicillin (AM), Dimetridazole (DMZ), Trimethoprim (TMP) were bought from the National Institute of Metrology in China, whereas Oxytetracycline (OTC), Caffeine (CF), and Diethyltoluamide (DEET) were acquired from the Toronto Research Chemicals Company in Canada.	2	1	Please add chemical reagents section and stated all chemical with brand specifications.	1	2	w14030367_perova	1
+These three GAC-UF systems were operated in parallel. Samples of the feedwater and effluent from three systems were taken simultaneously and measured once. The standard deviation was obtained by the detection over the full period of the experiment.	2	1	Regarding the replications, authors confirmed that replications of experiment were carried out. However, these results are not shown in the manuscript, how many replicated were carried out by experiment? Results seem to be related to a unique experiment. Please, clarify whether the results of this document are from a single experiment or from an average resulting from replications. If replicated were carried out, the use of average data is required as well as the standard deviation in the results and figures shown throughout the manuscript. In case of showing only one replicate explain why only one is shown and include the standard deviations.	1	2	w14030367_perova	1
+The figure is used as the only pattern in the revised manuscript.	2	1	Please revise your paper accordingly since some issue occurs on several spots in the paper. Please use Fig. or figure? It very confusing. Article should be in one pattern.	1	2	w14030367_perova	1
+The front sizes in the figure 1 have been increased. The quality of the figure 1 has been also improved.	2	1	Please provide high quality image of figure	1	2	w14030367_perova	1
+The style for units has been revised and unified.	2	1	Please use one style for units such as m3/h or m3h-1 Please revise your paper accordingly since some issue occurs on several spots in the paper.	1	2	w14030367_perova	1
+The comparison of previous research with existing research findings was added in the final part of the results and discussion. The added paragraph is as follows: PPCPs risks have been posing severe challenges to the safety of drinking water supply in rural areas due to the absence of the process with simple operation and maintenance as well as reliable performance. In this study, BAC coupling gravi-ty-driven UF was performed continuously, and the rejection performance of mem-brane filtration and BAC filtration both showed barriers for the conventional pollu-tants and PPCPs. Furthermore, this study indicated the respective contributions of BAC and UF, showing the role of the two-stage biofilm. Previous works involving BAC generally combined the ozonation with the BAC filter for treating the contaminants of emerging concern, eliminating a majority of PPCPs by more than 90% [34]. However, the regulation and maintenance of machines for ozone products are complicated, and the disinfection by-products will be newly generated in the effluent, which is incon-venient to use in rural areas [35]. In general, coagulation, filtration and single BAC units worked inefficiently and removed the detected PPCPs by less than 50%, as they were not hydrophobic [34, 36]. Hybrid membrane processes such as inline dosing of powdered activated carbon (PAC) prior to UF have already shown promising potential for the abatement of PPCPs; however, the inline dosing PAC is infeasible in rural areas [37]. In this study, the BAC prior to UF enhanced the biological activity by forming a two-stage biofilm system. Therefore, the integrated BAC-UF process can be considered as an economically and technically feasible approach to the decentralized and emer-gency drinking water treatment.	2	1	Please add a comparative profile section to compare your results and prove how it better than previous.	1	2	w14030367_perova	1
+The conclusion has been modified as suggested by reviewers. Please see lines 325-350 in the revised manuscript.	2	1	Section 4 should be renamed by Conclusion and Future perspectives. Conclusion section is missing some perspective related to the future research work, quantify main research findings, highlight relevance of the work with respect to the field aspect. In the present form conclusion is very weird.	1	2	w14030367_perova	1
+Two native English-speaking colleagues help us verify the manuscript. Hope the revised manuscript would be more satisfactory. We are so sorry to make reviewer’s reading uncomfortable. We have used an English Language Editing service to correct the grammatical and spelling errors and to make the expressions conform to correct scientific English (the Language Editing Certification is attached below).	2	1	To avoid grammar and linguistic mistakes, Major level English language should be thoroughly checked. Please revise your paper accordingly since several language issue occurs on several spots in the paper.	1	2	w14030367_perova	1
+We agree with the reviewer. The reference formatting has been corrected. Thanks very much for the patient review again.	2	1	Please follow the journal guidelines. Reference formatting need carefully revision. All must be consistent in one formate.	1	2	w14030367_perova	1
+Full names of BAC, UF, and PPCPs are used in the title and the abstract to show the results briefly. As for NH4+-N, NO2−-N, and NO3—N, the full names have been used instead of abbreviations in the abstract.	2	1	Authors should avoid abbreviations in the title and the abstract.	1	2	w14030367_perova	1
+The title has been revised to river water.	2	1	Authors may revise the title to include river water instead of surface water.	1	2	w14030367_perova	1
+The references format has been corrected.	2	1	References should be according to the journal format.	1	2	w14030367_perova	1
+We are very sorry for the confusion caused to the reviewers, the sentences have been revised from the words and Grammarly. The revised sentences are shown below: Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, col-loids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite [11, 12].	2	1	Line 44-45: Ultrafiltration (UF) as emerging technology, has been widely used to remove pollutants such as particles, colloids, bacteria and viruses, reducing the risk of water-borne diseases and …UF membranes cannot effectively rejected these soluble substances”. Please clarify, why ultrafiltration cannot remove the PPCPs since it can remove bacteria and viruses.	1	2	w14030367_perova	1
+The t-test results have been added in the revised Table1.	2	1	Please add t-test results in Table 1 for each parameter to understand the significant differences. Authors may provide data in the supporting information file.	1	2	w14030367_perova	1
+"The statistical t-test was evaluated for Figure 4, which was added in the text of ""3.2. Removal of nitrogen"" Section."	2	1	The reviewer suggests evaluating the statistical t-test for Figures 4.	1	2	w14030367_perova	1
+The t-test results have been added to the revised Table2. The concentration of Erythromycin was generally varied between 1047.14 ng L−1 and 2037.72 ng L−1.	2	1	t-test results should be included in Table Why is the standard deviation of Erythromycin showing a high value?	1	2	w14030367_perova	1
+Some symbols were overlapped in Figure 5h. The symbols near 85 days have been entirely shown by correcting Y-axis settings, such as Figures 5a and 5f.	2	1	Please look at the curve of BAC/UF; there was a symbol missing in near 85 days. Please show each symbol.	1	2	w14030367_perova	1
+The conclusion has been revised, including the style and sentences.	2	1	The conclusion may be revised.	1	2	w14030367_perova	1
+We agree with this suggestion. The abbreviations in the title have been deleted. Besides, a list of abbreviations has been added before the references.	4	1	Authors should avoid abbreviations in the title. Authors should add a list of abbreviations before the references.	3	2	w14030367_perova	1
+Thanks to the reviewer for the patient comment. The above mistakes have been revised.	4	1	Please avoid repeating the full name and abbreviation throughout the manuscript if you used the first-time abbreviation.	3	2	w14030367_perova	1
+1. 2. Thanks to this reviewer for the comment. Common waterborne viruses include Shigella, Vibrio cholerae, Poliovirus, adenovirus, and coxsackie virus, with sizes of 26 nm, 27 nm, 30 nm, 90 nm, and 30 nm, respectively [1-5]. The molecular weights of PPCPs are generally less than 1000 Da (about 1-2 nm), obviously smaller than the virus. The molecular weight cut off of UF membranes was an average 100 000 Da in this work, which was similar to the virus but larger than the PPCPs molecules. Karthik K, Dhanuskodi S, Gobinath C, et al. Multifunctional properties of microwave assisted CdO–NiO–ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities [J]. Journal of Materials Science: Materials in Electronics, 2018, 29(7): 5459-71. Song S, Liu Z, Zhou J, et al. An adjuvant compound that enhances immunogenicity at fractional doses of the Sabin-inactivated poliovirus vaccine (sIPV) with a long duration of protection in a rat model [J]. Journal of Medical Virology, 2019, 91(1): 14-21. Kim K, Choi J-W, Ma K, et al. Nanoisland-Based Random Activation of Fluorescence for Visualizing Endocytotic Internalization of Adenovirus [J]. Small, 2010, 6(12): 1293-9. Dourmashkin R R, Mccall S A, Dourmashkin N, et al. Virus-like particles and enterovirus antigen found in the brainstem neurons of Parkinson's disease [J]. F1000Res, 2018, 7: 302-. Tamano K, Aizawa S-I, Katayama E, et al. Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors [J]. The EMBO Journal, 2000, 19(15): 3876-87.	4	1	Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. Size exclusion is considered the primary removal mechanism for the UF. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite”. Sorry, but I don't see why UF can reject viruses and bacteria but not PPCPs. Is it true that the molecular weight of viruses and bacteria is more than that of PPCPs? Please elaborate.	3	2	w14030367_perova	1
+The information regarding the biological degradation of PPCPs has been added in the introduction. The revised sentences are shown below: In the BAC biofilters, the biotransformation and adsorption both contributed to the PPCPs removal. The activated carbon adsorbed PPCPs to the surface and interior, where microorganisms were suitable for growth. Under the long-term effect of high-concentration PPCPs, the dominant microorganisms in the biofilter were selected to survive. These microorganisms mostly transformed PPCPs into many segments and even directly mineralized them to CO2 [3, 14].	4	1	The authors should include some information regarding the biological degradation of PPCPs by microorganisms in the introduction section.	3	2	w14030367_perova	1
+“The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”. This claim is not supported by our research. To avoid misunderstandings for readers, we changed the claim and revised the sentence. The revised version is as follows: The combined process with UF may be another promising choice as an alternative to nanofiltration for removing PPCPs in rural areas. It can be seen in lines 54-55.	4	1	Line 53-55: Have the authors compared the BAC-UF performance to that of nanofiltration?	3	2	w14030367_perova	1
+The method of the t-tests has been explained in section 2.4 (lines 157-158) in the revised manuscript. The sample size (n) has also been added in Table 1 and Table 2.	4	1	I strongly suggest that the authors explain how they performed the t-tests. Please include the sample size (n) or degree of freedom (df).	3	2	w14030367_perova	1
+Thank you very much for your kind reminder. The values have been modified in the revised manuscript in yellow highlight.	4	1	Please show the t-value and p-value in a scientific view (e.g., 2.79E-4 would be P <.001).	3	2	w14030367_perova	1
+Thanks for the kind reminder. All the names of medicinal compounds not used have been deleted, including Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Roxithromycin (ROX), Penicillin-G (PEN-G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacin (IND), Metoprolol (METO), Propranolol (PRO), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), Sulpiride (SP), Sulfapyridine (SPN), Sulfamonomethoxine (SMM), Tetracycline (TC), Amoxicillin (AM), Dimetridazole (DMZ), Oxytetracycline (OTC) and DEET.	4	1	Line 161-176: Please delete the name of medicinal compounds not used for this study.	3	2	w14030367_perova	1
+The results and discussion has been supplemented in section 3.1. Please see lines 180-185 in the revised manuscript.	4	1	1) was observed in Fig. 1(c). Why is the DO concentration of BAC-effluent sometimes higher than the Raw water. Please explain it in the manuscript. The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent). Point 6: Why the same trend of increase or decrease in graphs (Fig. 1) was observed in Fig. 1(c).	3	2	w14030367_perova	1
+Thanks very much for this comment. With a turbulent current and dozens of kilometers in length, this river is located in the mountains of Foshan city. The PPCPs sludge at the bottom of the river never executes removal. Thanks for the suggestion from the reviewer. We will further precipitate the PPCPs sludge before the water treatment plant or excavate the PPCPs sludge from the river and study the effects on drinking water quality. The relevant results will be published in another paper in the future.	4	1	The same trend (Figure 2) was mainly due to the stable removal ability of BAC and UF for organics, causing the removal restriction. The periodic backwash (7days) of BAC caused sometimes the dissolved oxygen concentration of BAC-effluent higher than that of raw water. After the gas scrubbing and the hydraulic backwashing, the dissolved oxygen detection of the effluent was carried out, resulting in the above results for dissolved oxygen. Besides, the t-test was used and proved the significant difference between BAC-Effluent and BAC/UF-effluent. Response 6: Thanks very much for this comment.	3	2	w14030367_perova	1
+Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.	2	1	There is no sampling procedure, depth, or random selection of the soils evaluated.	1	2	w14081258_makarova	1
+It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615. The values obtained of I DW and steric hindrance are not explained.	2	1	Is the explanation of χ is for molecular connectivity indices or branching index?	1	2	w14081258_makarova	1
+The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications. Shiyu Wang, Fei Liu*, Wenyong Wu*, Yaqi Hu, Renkuan Liao, Gaoting Chen, Jiulong Wang, Jialin Li. Migration and health risks of nonylphenol and bisphenol A in soil-winter wheat systems with long-term reclaimed water irrigation. Ecotoxicology and Environmental Safety 158 (2018) 28–36. Shiyu Wang，Wenyong Wu*，Fei Liu, Xiaoou Li. Sorption and desorption behaviours of 4-nonylphenol on reclaimed water-irrigated soils. Environmental Engineering Science. (2019) 36 (9) : 1100-1111。 王世玉，刘菲*，刘玉龙，陈亮. 气相色谱-质谱法检测地下水中12种对壬基酚同分异构体.分析化学，2013，41（11）：1699-1703. 王世玉，刘菲*，吴文勇，尹世洋，刘玉龙，陈亮，张伟，陈会会.影响12种壬基酚同分异构体液液萃取效率的因素研究. 岩矿测试，2014，33（4）：570-577.	2	1	If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.	1	2	w14081258_makarova	1
+It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.	2	1	The basic physi-chemical parameters of soils should be given.	1	2	w14081258_makarova	1
+Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu. Spatial distribution and migration of nonylphenol in groundwater following long-term wastewater irrigation. Journal of Contaminant Hydrology,2015,177-178（June–July）:85-92. I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.	2	1	The irrigation water physi-chemical parameters should be given.	1	2	w14081258_makarova	1
+It has been given in Table1.	2	1	The kinect models should be given.	1	2	w14081258_makarova	1
+Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.	2	1	Figure 1 should be revised according to kinect model.	1	2	w14081258_makarova	1
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.	2	1	I strongly recommend the authors to analysis the biomass of the soil samples.	1	2	w14081258_makarova	1
+Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.	4	1	"I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: ""The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars"" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?"	3	2	w14081258_makarova	1
+Accepted and revised in L204. Figure 1 has been changed in the manuscript.	4	1	Figure 1 legends of axis need font size larger, please make readable the figuresR：	3	2	w14081258_makarova	1
+Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.	2	1	There is no sampling procedure, depth, or random selection of the soils evaluated.	1	2	w14081258_perova	1
+It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615.	2	1	Is the explanation of χ is for molecular connectivity indices or branching index?	1	2	w14081258_perova	1
+Accepted and revised in L157-160. Common topological indices, such as κ shape indices and information indices were also calculated by Molconn-Z (version 4.12S, eduSoft, La Jolla, CA). The detailed list and definitions can be found in the software user’s guide [35], and the definitions can also be found in Todeschini et al. [36]. The results of other common topological indices calculated by Molconn-Z were shown below as well. But in this study, only the steric index (R2 = 0.82 for CN soil; R2 = 0.86 for FN soil) and IDWbar (R2 = 0.83 for CN soil; R2 = 0.94 for FN soil) have a better relationship with the half lives of the isomers, which were mentioned in the Results and Discussion part. The results of the topological indices of the isomers [35] Hall, L. H.; Kellogg, G. E.; Haney, D. N. (2008). Software Package for Molecular Topology Analysis User’S Guide. [36] Todeschini, R.; Consonni, V.; Mannhold, R.; Kubinyi, H.; Folkers, G. (2009). Molecular Descriptors for Chemoinformatics; Wiley: New York.	2	1	The values obtained of I DW and steric hindrance are not explained.	1	2	w14081258_perova	1
+The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications.	2	1	The more critical issue lies in the analytical methodology used for isomers quantification. First of all, figure one is a chromatogram, not a spectrum. The chromatogram is not well resolved, i.e., there is no separation among the different analytes determined. For example, look at peaks 4, 5, and 6. There is no resolution among the isomers, the same for NP 7 and 8 and NP 8 and 10; thus, it is impossible to quantify them. If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.	1	2	w14081258_perova	1
+It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.	2	1	The basic physi-chemical parameters of soils should be given.	1	2	w14081258_perova	1
+I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.	2	1	The irrigation water physi-chemical parameters should be given.	1	2	w14081258_perova	1
+It has been given in Table1.	2	1	The kinect models should be given.	1	2	w14081258_perova	1
+Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.	2	1	Figure 1 should be revised according to kinect model.	1	2	w14081258_perova	1
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.	2	1	I strongly recommend the authors to analysis the biomass of the soil samples.	1	2	w14081258_perova	1
+Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.	2	1	"I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: ""The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars"" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?"	1	2	w14081258_perova	1
+Accepted and revised in L204. Figure 1 has been changed in the manuscript.	2	1	Figure 1 legends of axis need font size larger, please make readable the figuresR：	1	2	w14081258_perova	1
+However, on pages 7 - 9 the authors admit that they divert from the CIRA procedure by identifying the outcome in advance, assigning qualitative values to preferences, assuming weights, utility factors, and initial values, and considering several stakeholders as risk owners.			As a separate recommendation, the paper needs smoother transitions between topics on pages 2-7. The authors should add one or two lines smoothing the flow for the reader, rather than relying on subject headings.	1	2	admsci5030125_boyarkin	0
+This should change to “to manage the risk of human factors or to risk management of human factors”.			The paper is on the one hand quite verbose, but with respect to the peer review incomplete. Possible risks in peer review are many more than the ones listed in Table 3.	1	2	admsci5030125_boyarkin	0
+I have tried to accomodate your comments to the best of my ability.			The paper can become stronger if it limits itself to the Peer Review Ring Incident and will look at the various aspects of that scenario including risk reducing measures.	1	2	admsci5030125_boyarkin	0
+I have tried to accomodate your comments to the best of my ability.			It didn't become clear to me as a reader where the specific results of the 13 steps of the CIRA application are and where these are described. What are the conclusions about the root cause? Or are there multiple root causes? And what definition of root cause is used?	1	2	admsci5030125_boyarkin	0
+Kind regards, Gaute Wangen Second Round of Evaluation Round 2: Despite the article even became longer, the line of thought is now clear.			there is not enough explanation about the CIRA and the person(s) who developed this method. Also there is not enough clarification and elaboration on the differences of your work on CIRA and your customizations comparing to the original methodology.	1	2	admsci5030125_boyarkin	0
+I believe you mistakenly have the caption of figure 1 in here.			There are some typos, punctuations and grammatical errors in the paper,	1	2	admsci5030125_boyarkin	0
+You brought up a very interesting topic/subject of research in this study/paper, which I believe it is of interest and use to many journals for their peer review process.			There are some consistency issues in your writing. This does not look professional in scientific papers.	1	2	admsci5030125_boyarkin	0
+Round 2: Author Response to Reviewer 3 Dear reviewer, Thank you for the read-through of my manuscript.			There is a need for some editing; e.g. grammatical and punctuation corrections, elements in the text.	3	2	admsci5030125_boyarkin	0
+The main contribution of the paper and its deliverable are described.			Given the authors stated goal of promoting the CIRA technique and approach to demonstrating its uses, I would recommend re-formatting the paper as a pedagogical tool. Rather than claiming to validate the method, focus on a detailed step by step examination of its implementation using a known outcome.	1	2	admsci5030125_makarova	0
+As an editor I would think twice if a young scientist whom I don't know offers her./himself as a reviewer and I would certainly ask for recommendations.			The paper is on the one hand quite verbose, but with respect to the peer review incomplete.	1	2	admsci5030125_makarova	0
+R5 Round 1: Author Response to Reviewer 3 Dear Reviewer 3, Thank you for taking the time to review my work and provide very good comments on my work.			The paper can become stronger if it limits itself to the Peer Review Ring Incident and will look at the various aspects of that scenario including risk reducing measures.	1	2	admsci5030125_makarova	0
+I have included all your suggested changes together with a major spellcheck.			What are the conclusions about the root cause? Or are there multiple root causes? And what definition of root cause is used?	1	2	admsci5030125_makarova	0
+The three dots represent an incomplete sentence.			Also there is not enough clarification and elaboration on the differences of your work on CIRA and your customizations comparing to the original methodology.	1	2	admsci5030125_makarova	0
+I believe you mistakenly have the caption of figure 1 in here.			There is a need for some editing; e.g. grammatical and punctuation corrections, elements in the text.	3	2	admsci5030125_makarova	0
+In both cases, Principle-Agent assumptions apply, but not because both are based on for-profit assumptions.			the third sector, its definition and description is missing. I would not have thought of this as a major issue unless Evers and Laville (2004) The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector. Maybe taken for granted here? The least we should expect is a definition.	1	2	admsci5030148_devetyaro	0
+Round 1: Author Response to Reviewer 3 I copied and pasted the reviewer commentary, below.			the case description and analysis is based only on media accounts - no actual first hand information to inform the analysis. I find this less than satisfying. It should be possible to interview a few key individuals about the case to make sure the medis picture is corrected where this is needed. Media does not always get it right. There is not enough evidence to prove anything about CEC.	1	2	admsci5030148_devetyaro	0
+There is not enough evidence to prove anything about CEC.			I also have problems with the depth of the discussion, and I feel the conclusion does not match the promises of the abstract.	1	2	admsci5030148_devetyaro	0
+I do not agree with the author for several reasons.			Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders)	1	2	admsci5030148_devetyaro	0
+I do not expect the complete picture, but should have comments on limitations and to what degree the conclusions and learnings are possible to generalize.			Bottom of page 9: Missing reference to Co-operative principles by Rochdale Society.	1	2	admsci5030148_devetyaro	0
+Your comments have made this paper significantly better.			USA context concerning cooperatives (upper half page 10) should be highlighted even more for readers to be able to compare to their own context (for instance in Europe)	1	2	admsci5030148_devetyaro	0
+The text itself is now almost finished, I only have a few minor comments: -			The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?	1	2	admsci5030148_devetyaro	0
+Thanks so much for your thoughtful commentary!			are references 27 and 33 used in the text? (could not find them) Good luck in further research and writing on this subject.	1	2	admsci5030148_devetyaro	0
+There is, however, some common practices, but not for the reasons described.			The literature part is improved by changing the headline, putting it in the right setting. The change illustrates how headlines can change the apperance of the text contents. However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles? If they are - the connection need explainnig. If they are not, the main headling Chapter 2 should be reformulated again.	3	2	admsci5030148_devetyaro	0
+Effective governance in nonprofit organizations: a literature based multiple stakeholder approach, European Management Journal, 32, 2: 223-243.			I still miss a source reference at the bottom of page 9 (Rochdale Society).	3	2	admsci5030148_devetyaro	0
+The style is more journalistic and free opinion than rigourous research.			On top of page 11 there is still no examples of adversarial effects removed by co-operative.	3	2	admsci5030148_devetyaro	0
+The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			The comments added in the beginning of Chapter 3 on methodology are important and improves the text a lot. The author openly acknowledges that the basis for a complete picture is lacking. I do not expect the complete picture, but should have comments on limitations and to what degree the conclusions and learnings are possible to generalize.	3	2	admsci5030148_devetyaro	0
+), Nonprofit Governance: Innovative Perspectives and Approaches.			"Conclusion: middle of page 16: ""we cannot know the motivations behind.."" Have you tried to talk to them? I believe it is possible to get to know."	3	2	admsci5030148_devetyaro	0
+To address these subtlies see: Turnbull (2002) A New Way to Govern Cornforth (2004) The Governance of Associations and Mutuals: A Paradox Perspective Ridley-Duff (2007) Communitarian Perspectives on Social Enterprise			"Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"	3	2	admsci5030148_devetyaro	0
+Just as the wealthy entrepreneur seeks to control the private enterprise through shareholdings, so the wealthy philanthropist seeks to control the non-profit enterprise through trust law.			Furthermore, there is no discussion of principal-agent literature in a third sector context (see, for example, Jegers, 2009), which is quite surprising since the proposition to go beyond the traditional principal-agent literature is one of the main ideas of the manuscript.	1	2	admsci5030148_devetyaro	0
+Given the case study, I would focus on the governance of co-operatives (Cornforth, 2004).			Furthermore, attempts have already been made to broaden the principal-agent approach in nonprofit organizations (Ben-Ner et al., 2012; Van Puyvelde et al., 2012, Coule, 2015), leading me to the question what this manuscript actually adds to the literature.	1	2	admsci5030148_devetyaro	0
+R4 Good luck in further research and writing on this subject.			In the introduction, the author also mentions underdeveloped modes of stakeholder governance in the third sector. I do not agree with this view. Several stakeholder governance frameworks for third sector organizations have already been developed in the literature (Young, 2011; Van Puyvelde et al., 2012; Wellens and Jegers, 2014).	1	2	admsci5030148_devetyaro	0
+Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			The author uses “stakeholder-controlled firm”, “third sector organizations”, “nonprofit organizations”, “social enterprise”, and “co-operative” without clearly defining these types and mentioning which nonprofit type is actually the focus of the study.	1	2	admsci5030148_devetyaro	0
+The author openly acknowledges that the basis for a complete picture is lacking.			I am also not convinced by the definition of co-operatives mentioned in the paper.	1	2	admsci5030148_devetyaro	0
+METHODOLOGY This is weak, perhaps too weak for an academic paper.			Furthermore, given that the world of third sector organizations is very rich and heterogeneous, one should be careful by discussing third sector organizations in general, especially since the case study seems to focus on co-operative enterprises. Instead of being vague and using a mixed terminology, the manuscript should clearly distinguish which nonprofit type is the subject of the paper, and be very careful in generalizing the findings of this study to third sector governance, which mistakenly happens in the conclusion (p. 15). Given the case study, I would focus on the governance of co-operatives	1	2	admsci5030148_devetyaro	0
+Furthermore, attempts have already been made to broaden the principal-agent approach in nonprofit organizations (Ben-Ner et al., 2012; Van Puyvelde et al., 2012, Coule, 2015), leading me to the question what this manuscript actually adds to the literature.			The literature review is completely missing.	1	2	admsci5030148_devetyaro	0
+The governance of cooperatives and mutual associations: A paradox perspective.			The literature review is completely missing.	1	2	admsci5030148_devetyaro	0
+COMMENT #1 This is a significantly improved version of the manuscript.			Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.	1	2	admsci5030148_devetyaro	0
+COMMENT #3: On top of page 11 there is still no examples of adversarial effects removed by co-operative.			I believe that Cornforth’s paper (2004) might be a good starting point.	1	2	admsci5030148_devetyaro	0
+Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.			In addition, a more extensive literature review and comparison with previous principal-agent literature and stakeholder governance models (especially Van Puyvelde et al., 2012 and Coule, 2015) may give the manuscript a more solid theoretical foundation. As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.	1	2	admsci5030148_devetyaro	0
+(2012) and the idea that is necessary to capture the broader stakeholder environment in a democratic way is also not new (Coule, 2015).			The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.	1	2	admsci5030148_devetyaro	0
+I would not have thought of this as a major issue unless Evers and Laville (2004) The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector.			The characterisation of third sector governance is too crude. For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors. The comments at the start of the literature review might apply to foundations and charities, but are less true of association and mutuals (let alone cooperatives). Also, where there are accounting regulations (like SORP), there are mechanism that keep associations and charities accountable to their donors.	1	2	admsci5030148_devetyaro	0
+The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation.			Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.	1	2	admsci5030148_devetyaro	0
+Nonprofit and Voluntary Sector Quarterly 44(1): 75-97.			Secondly, I just can't agree that third sector governance is derived from for-profit principles. It is - mostly - derived from philanthropic principles (or mutual principles inside the social economy). There is, however, some common practices, but not for the reasons described. Just as the wealthy entrepreneur seeks to control the private enterprise through shareholdings, so the wealthy philanthropist seeks to control the non-profit enterprise through trust law. In both cases, Principle-Agent assumptions apply, but not because both are based on for-profit assumptions. These apply in both cases because funding is sought from wealthy providers with the power to frame laws that require their investees to use the money for the purposes stimpulated by them. In the social economy (associations, mutuals and cooperatives) the legal structures and accountabilities are different (and are captured well in the paper). Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal structures and accountability mechanisms reflect this.	1	2	admsci5030148_devetyaro	0
+The text itself is now almost finished, I only have a few minor comments: -			This is weak, perhaps too weak for an academic paper. Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable. The use of media sources, while not ideal, might be adequate if the author(s) can show systematic and comprehensive collection of them. However, the credibility of the paper and findings would be enormously enhanced is some primary interview data could be collected and presented. Is there a way to interview members who participated in action (or access sources written by them)?	1	2	admsci5030148_devetyaro	0
+The governance of cooperatives and mutual associations: A paradox perspective.			The framework for this section exists but feels a bit superficial and under-developed. I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).	1	2	admsci5030148_devetyaro	0
+"COMMENT #6: Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"			Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification. Stick to commenting on the usefulness and value of ODPs, and draw out any theoretical contributions regarding the design principles themselves, or their value as a theoretical perspective for governance research. Some well grounded comments on their use in other governance research are merited right at the end.	1	2	admsci5030148_devetyaro	0
+Nonprofit governance research: Limitations of the focus on boards and suggestions for new directions.			I do not understand the purpose of the paper.	1	2	admsci5030148_devetyaro	0
+In addition, a more extensive literature review and comparison with previous principal-agent literature and stakeholder governance models (especially Van Puyvelde et al., 2012 and Coule, 2015) may give the manuscript a more solid theoretical foundation.			The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.	1	2	admsci5030148_devetyaro	0
+Nonprofit and Voluntary Sector Quarterly 40(4): 608-633.			The style is more journalistic and free opinion than rigourous research.	1	2	admsci5030148_devetyaro	0
+The author openly acknowledges that the basis for a complete picture is lacking.			Public enterprises, third sector cooperatives are all mixedup.	1	2	admsci5030148_devetyaro	0
+Similarly, in the introduction, it is stated that governance research on third sector organizations predominantly fixates analysts on the board-management dynamic.			The conceptual framework is not clearly presented and I do not see what are the contributions except some free statements at the end not really related to the analysis.	1	2	admsci5030148_devetyaro	0
+However, a solid foundation for this argument is missing.			the third sector, its definition and description is missing. The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector.	1	2	admsci5030148_makarova	0
+Furthermore, attempts have already been made to broaden the principal-agent approach in nonprofit organizations (Ben-Ner et al., 2012; Van Puyvelde et al., 2012, Coule, 2015), leading me to the question what this manuscript actually adds to the literature.			the case description and analysis is based only on media accounts - no actual first hand information to inform the analysis.	1	2	admsci5030148_makarova	0
+Is there a way to interview members who participated in action (or access sources written by them)?			I also have problems with the depth of the discussion, and I feel the conclusion does not match the promises of the abstract.	1	2	admsci5030148_makarova	0
+R8 Note that the paper no longer claims this to be a case study, and instead positions itself as a discussion paper, performing a secondary archival analysis of media accounts.			Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders)	1	2	admsci5030148_makarova	0
+Round 1: Author Response to Reviewer 1 I copied and pasted the reviewer commentary, below.			Bottom of page 9: Missing reference to Co-operative principles by Rochdale Society.	1	2	admsci5030148_makarova	0
+Some well grounded comments on their use in other governance research are merited right at the end.			USA context concerning cooperatives (upper half page 10) should be highlighted even more for readers to be able to compare to their own context (for instance in Europe)	1	2	admsci5030148_makarova	0
+As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.			The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?	1	2	admsci5030148_makarova	0
+Introducing the Ostrom Design Principles: From Unitarist to More Pluralist Governance Arrangements 2.2.The Institutional Design of the Co-operative Institutional Model COMMENT #2: I still miss a source reference at the bottom of page 9 (Rochdale Society).			are references 27 and 33 used in the text? (could not find them) Good luck in further research and writing on this subject.	1	2	admsci5030148_makarova	0
+Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles? If they are - the connection need explainnig. If they are not, the main headling Chapter 2 should be reformulated again.	3	2	admsci5030148_makarova	0
+Second, I note in the paper that I am not making the claim that the co-operative model is part of the third sector.			I still miss a source reference at the bottom of page 9 (Rochdale Society).	3	2	admsci5030148_makarova	0
+Effective governance in nonprofit organizations: a literature based multiple stakeholder approach, European Management Journal, 32, 2: 223-243.			On top of page 11 there is still no examples of adversarial effects removed by co-operative.	3	2	admsci5030148_makarova	0
+Young DR. (2011).The prospective role of economic stakeholders in the governance of nonprofit organizations.			The author openly acknowledges that the basis for a complete picture is lacking. I do not expect the complete picture, but should have comments on limitations and to what degree the conclusions and learnings are possible to generalize.	3	2	admsci5030148_makarova	0
+The style is more journalistic and free opinion than rigourous research.			"Conclusion: middle of page 16: ""we cannot know the motivations behind.."" Have you tried to talk to them? I believe it is possible to get to know."	3	2	admsci5030148_makarova	0
+The two major issues are: - the third sector, its definition and description is missing.			"Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"	3	2	admsci5030148_makarova	0
+The author uses “stakeholder-controlled firm”, “third sector organizations”, “nonprofit organizations”, “social enterprise”, and “co-operative” without clearly defining these types and mentioning which nonprofit type is actually the focus of the study.			The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation. Similarly, in the introduction, it is stated that governance research on third sector organizations predominantly fixates analysts on the board-management dynamic.	1	2	admsci5030148_makarova	0
+- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			In the abstract, the author also states that the over-reliance on the principal-agent model introduces two challenges. However, a solid foundation for this argument is missing.	1	2	admsci5030148_makarova	0
+METHODOLOGY This is weak, perhaps too weak for an academic paper.			In the introduction, the author also mentions underdeveloped modes of stakeholder governance in the third sector. I do not agree with this view. Several stakeholder governance frameworks for third sector organizations have already been developed in the literature (Young, 2011; Van Puyvelde et al., 2012; Wellens and Jegers, 2014).	1	2	admsci5030148_makarova	0
+Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal structures and accountability mechanisms reflect this.			The author uses “stakeholder-controlled firm”, “third sector organizations”, “nonprofit organizations”, “social enterprise”, and “co-operative” without clearly defining these types and mentioning which nonprofit type is actually the focus of the study.	1	2	admsci5030148_makarova	0
+As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.			I am also not convinced by the definition of co-operatives mentioned in the paper.	1	2	admsci5030148_makarova	0
+For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors.			Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.	1	2	admsci5030148_makarova	0
+Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.			The characterisation of third sector governance is too crude.	1	2	admsci5030148_makarova	0
+For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors.			Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.	1	2	admsci5030148_makarova	0
+COMMENT #3: On top of page 11 there is still no examples of adversarial effects removed by co-operative.			I just can't agree that third sector governance is derived from for-profit principles.	1	2	admsci5030148_makarova	0
+Instead of being vague and using a mixed terminology, the manuscript should clearly distinguish which nonprofit type is the subject of the paper, and be very careful in generalizing the findings of this study to third sector governance, which mistakenly happens in the conclusion (p. 15).			Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.	1	2	admsci5030148_makarova	0
+Just as the wealthy entrepreneur seeks to control the private enterprise through shareholdings, so the wealthy philanthropist seeks to control the non-profit enterprise through trust law.			The framework for this section exists but feels a bit superficial and under-developed. I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).	1	2	admsci5030148_makarova	0
+Thank you very much for your helpful commentary!			Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.	1	2	admsci5030148_makarova	0
+- Bottom of page 9: Missing reference to Co-operative principles by Rochdale Society.			I do not understand the purpose of the paper.	1	2	admsci5030148_makarova	0
+Round 1: Author Response to Reviewer 3 I copied and pasted the reviewer commentary, below.			The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.	1	2	admsci5030148_makarova	0
+Nonprofit and Voluntary Sector Quarterly 41(6): 1116-1135.			The style is more journalistic and free opinion than rigourous research.	1	2	admsci5030148_makarova	0
+- The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?			Public enterprises, third sector cooperatives are all mixedup.	1	2	admsci5030148_makarova	0
+The text itself is now almost finished, I only have a few minor comments: - The literature part is improved by changing the headline, putting it in the right setting.			The conceptual framework is not clearly presented and I do not see what are the contributions except some free statements at the end not really related to the analysis.	1	2	admsci5030148_makarova	0
+It seems that the most prominent or successful cases have been selected, at least for Germany and France.			the analysis of the data is weak, and the paper remains largely descriptive. Would suggest to use these case studies as a source of ideas on how policies and cluster initative management migth relate to each other; that would make this a more interesting paper.	1	2	admsci5040213_boyarkin	0
+According to the definition on the top of page 4, the term denotes a R9 cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper.			The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included.	1	2	admsci5040213_boyarkin	0
+(…) A conceptual and/or theoretical perspective is needed to shed light at the differences found.			"There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_boyarkin	0
+P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.			The language the authors use is here a bit imprecise - they seem to be talking about the management of cluster initiatives, not about the management of the policies themselves (which is more a question of how the program oversight is organized within government; also an interesting but different question).	1	2	admsci5040213_boyarkin	0
+P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.			I would encourage you to have a native speaker look at it, mainly to clarify the language in some places.	3	2	admsci5040213_boyarkin	0
+Remaining Comments Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators.			Content-wise to me the key observation is the - potentially systematic - connection between the way cluster policies arestructure and the organization of cluster initiatives. This is not a big suprise, and it doesn't really answer the question asto which model is better (only that there are internally consistent models that are different). But it is still a usual observation to make.	3	2	admsci5040213_boyarkin	0
+Round 2: Author Response to Reviewer 1 Thank you for your comments.			the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2. It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices. The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters. As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope.	1	2	admsci5040213_boyarkin	0
+• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2. It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices. The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters. As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope.	1	2	admsci5040213_boyarkin	0
+‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters.			‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance. Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp. regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.	1	2	admsci5040213_boyarkin	0
+KIESE, M., 2009: National styles of cluster promotion: Cluster policies between variety and convergence.			At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.	1	2	admsci5040213_boyarkin	0
+In the following, we will reply to each of your comments, using blue letters for our responses.			A conceptual and/or theoretical perspective is needed to shed light at the differences found. in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up. The authors should include a discussion on how these countries differ e.g.	1	2	admsci5040213_boyarkin	0
+"There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."			The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies. According to the definition on the top of page 4, the term denotes a cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper. Even so, the term ‘intellectual’ does not look fully appropriate here, which may be due to its translation from a Japanese original. ‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters. At the least, it would appear sound to qualify these six cases as science-based clusters, but then this would apply to the biotech industry and all its clusters in general.	1	2	admsci5040213_boyarkin	0
+However, it remains rather descriptive and does not systematically attempt to explain the differences found between the three countries.			The choice of clusters is generally well explained on pp. 2-3. When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!	1	2	admsci5040213_boyarkin	0
+Since this may be managed by the editorial office, I do not see the need for another review.			When explaining the choice of regional cases, the term ‘representative’ appears misleading. It seems that the most prominent or successful cases have been selected, at least for Germany and France. How can these be ‘representative’ for whatever population of biotech clusters in these countries?	1	2	admsci5040213_boyarkin	0
+In: Journal of Economic Issues, 41(2): 391-398.			When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number. Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e. representatives of the respective national cluster programmes, have not been surveyed as well.	1	2	admsci5040213_boyarkin	0
+Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.			When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number. Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e. representatives of the respective national cluster programmes, have not been surveyed as well.	1	2	admsci5040213_boyarkin	0
+Freeman, C., 1988: Japan: A new National System of Innovation?			some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_boyarkin	0
+P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.			sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between. Compared to the Japanese cases, the French and German cases are clearly more bottom-up, but they still display clear – if not dominant – elements of top-down governance and public agency. This dichotomy should be refined and the classification of cases qualified as, e.g., ‘relatively bottom-up’.	1	2	admsci5040213_boyarkin	0
+• Finally, the paper is static as it does not account for cluster evolution and (policy) learning over time.			For the German cases, the authors may have underestimated public agency, since they did not investigate the role of state governments (Bavaria, Baden-Württemberg) within the country’s system of multilevel governance, which is key to understand cluster policies in Germany (cf. KIESE 2013). Furthermore, the state government of Bavaria may be a regional government, but it is certainly not a local authority (like the city of Munich; page 15, line 29) in Germany’s system of multilevel governance.	1	2	admsci5040213_boyarkin	0
+• Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.			Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment. This is probably the result of a rather weak empirical basis, provided it draws on interviews with a total of six cluster managers. Critical issues such as the cluster management organisation’s budget and finance have not been addressed. The relevance of the information presented here for the following discussion and conclusion remain far from clear. In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.	1	2	admsci5040213_boyarkin	0
+Since this may be managed by the editorial office, I do not see the need for another review.			Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment. This is probably the result of a rather weak empirical basis, provided it draws on interviews with a total of six cluster managers. Critical issues such as the cluster management organisation’s budget and finance have not been addressed. The relevance of the information presented here for the following discussion and conclusion remain far from clear. In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.	1	2	admsci5040213_boyarkin	0
+"There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."			As also elaborated above, the conclusion should go beyond a mere summary to include methodological reflection, implications for policy (learning), as well as a more extensive discussion of further research perspectives.	1	2	admsci5040213_boyarkin	0
+It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.			Page 3, line 3: The authors might want to link to the literature on differentiated knowledge bases, which assigns an analytical (science-based) knowledge base to the biotech industry (cf. ASHEIM ET AL. 2011).	1	2	admsci5040213_boyarkin	0
+But the analysis of the data is weak, and the paper remains largely descriptive.			Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references. If ‘few’ means ‘no’, then please say so straightaway.	1	2	admsci5040213_boyarkin	0
+• Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.			Page 6, line 25: I would suggest softening the statement that matched funding ‘prevents moral hazard and crowding out’. It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.	1	2	admsci5040213_boyarkin	0
+The authors conclude that they found ‘consistent combinations of the types of national policies, local clusters, and cluster management’.			Section 2 contains a number of “expectations”, which could be flagged out and numbered as hypotheses. Taking these up in the empirical discussion could help the reader’s orientation.	1	2	admsci5040213_boyarkin	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			Page 8, lines 1-2: This is interesting indeed. Is there comparable evidence on policy learning between Germany and France? Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.	1	2	admsci5040213_boyarkin	0
+Taking these up in the empirical discussion could help the reader’s orientation.			Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.	1	2	admsci5040213_boyarkin	0
+For instance, what does it mean if a regional cluster organisation is pre-dating the national policy programme it is supposed to implement, as evident from some of the cases?			Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.	1	2	admsci5040213_boyarkin	0
+http://revel.unice.fr/eriep/index.html?id=3495, accessed June 13, 2015.			Section 3.3: There is a little bit of literature on French national cluster policy that deserves a brief review or at least reference here	1	2	admsci5040213_boyarkin	0
+• Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.			Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.	1	2	admsci5040213_boyarkin	0
+in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up.			Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’? If the aim of public policy is to grow a cluster around a research organisation or hospital, this does not seem to produce any meaningful outcomes as yet.	1	2	admsci5040213_boyarkin	0
+I would still like to see a qualification stating that R16 private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.			Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’. This should be backed up with evidence (data) and preferably rephrased, such as ‘recognised research universities’, or ‘elite’ with reference to the German federal government’s programme of university excellence (Exzellenzinitiative).	1	2	admsci5040213_boyarkin	0
+As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing.			Page 15, line 27: Repetition – the IZB incubator was already mentioned on line 9.	1	2	admsci5040213_boyarkin	0
+In: Environment and Planning C, 28(6): 1063-1082.			Section 4.5: Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that KOSCHATZKY (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.	1	2	admsci5040213_boyarkin	0
+"There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."			The layout of the tables may be improved, especially the spacing of the text.	1	2	admsci5040213_boyarkin	0
+The revised version went through a professional, native English editing.			The paper contains funding information in Yen and Euros. This should be harmonised, or preferably conversions given in brackets or footnotes for the reader’s convenience.	1	2	admsci5040213_boyarkin	0
+Round 1: Author Response to Reviewer 2 We are very grateful for your detailed, insightful comments and suggestions, which help improve our manuscript significantly.			Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators. If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences. This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation. A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).	3	2	admsci5040213_boyarkin	0
+o P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably R15 taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?			Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique. I would still like to see a qualification stating that private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.	3	2	admsci5040213_boyarkin	0
+This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation.			P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.	3	2	admsci5040213_boyarkin	0
+Round 1: Referee Report for Administrative Sciences Management of cluster policies for innovation: Comparative case studies of Japanese, German, and French biotechnology clusters Manuscript no.			P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.” In response, the authors now cite this paper in footnote 14 in Section 3.2. to support the view that BioValley has hardly had any impact in terms of intensifying cross-border interaction. This reformulation is incorrect as the Koschatzky (2000) paper pre-dated the BioValley initiative and does not look at INTERREG either. Unfortunately, my claim regarding the limited impact of BioValley on cross-border networking draws on conversations with practitioners and a field trip to the region, but cannot be supported by a reference as far as I know. I would therefore suggest to drop this footnote altogether.	3	2	admsci5040213_boyarkin	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			P. 16: Sources should be stated underneath table 2.	3	2	admsci5040213_boyarkin	0
+When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!			P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.	3	2	admsci5040213_boyarkin	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?	3	2	admsci5040213_boyarkin	0
+regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.			But the analysis of the data is weak, and the paper remains largely descriptive. Would suggest to use these case studies as a source of ideas on how policies and cluster initative management migth relate to each other; that would make this a more interesting paper.	1	2	admsci5040213_devetyaro	0
+• Section 3.3: There is a little bit of literature on French national cluster policy that deserves a brief review or at least reference here (e.g., BRETTE/CHAPPOZ 2007, DURANTON ET AL. 2010, LONGHI/ROCHHIA 2013).			"The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included. There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_devetyaro	0
+As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope.			"The conceputal framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included. There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti). There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."	1	2	admsci5040213_devetyaro	0
+1992: National Systems of Innovation: Towards a Theory of Innovation and Interactive Learning.			The language the authors use is here a bit imprecise - they seem to be talking about the management of cluster initiatives, not about the management of the policies themselves (which is more a question of how the program oversight is organized within government; also an interesting but different question).	1	2	admsci5040213_devetyaro	0
+Such limitations do not necessarily disqualify the paper, but should be discussed reflexively in the paper’s conclusions.			Content-wise to me the key observation is the - potentially systematic - connection between the way cluster policies arestructure and the organization of cluster initiatives. This is not a big suprise, and it doesn't really answer the question asto which model is better (only that there are internally consistent models that are different). But it isstill a usual observation to make.	3	2	admsci5040213_devetyaro	0
+In: Environment and Planning C, 28(6): 1063-1082.			However, the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2.	1	2	admsci5040213_devetyaro	0
+The varieties of capitalism (VoC) concept developed by HALL/SOSKICE (2011) may be a suitable concept here, as it has been applied to cluster policies in the U.S. and Germany by STERNBERG ET AL. (2010).			As scholars, the authors should challenge this technocratic faith of policy-makers and practitioners, although evaluation is beyond this paper’s scope. The performance of clusters is also affected by many exogenous influences not captured in the simple model.	1	2	admsci5040213_devetyaro	0
+• Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’?			Furthermore, ‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance. Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp. regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.	1	2	admsci5040213_devetyaro	0
+• Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.			At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.	1	2	admsci5040213_devetyaro	0
+A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).			However, it remains rather descriptive and does not systematically attempt to explain the differences found between the three countries.	1	2	admsci5040213_devetyaro	0
+It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number.			According to the definition on the top of page 4, the term denotes a cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper. Even so, the term ‘intellectual’ does not look fully appropriate here, which may be due to its translation from a Japanese original. ‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters. At the least, it would appear sound to qualify these six cases as science-based clusters, but then this would apply to the biotech industry and all its clusters in general.	1	2	admsci5040213_devetyaro	0
+In: European Review of Industrial Economics and Policy (5).			When explaining the choice of regional cases, the term ‘representative’ appears misleading.	1	2	admsci5040213_devetyaro	0
+Such limitations do not necessarily disqualify the paper, but should be discussed reflexively in the paper’s conclusions.			When explaining the choice of regional cases, the term ‘representative’ appears misleading.	1	2	admsci5040213_devetyaro	0
+I would still like to see a qualification stating that R16 private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.			When outlining their research methodology, the authors should state precisely how many interviews they conducted. It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number.	1	2	admsci5040213_devetyaro	0
+representatives of the respective national cluster programs, have not been surveyed as well.			Furthermore, some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_devetyaro	0
+R12 • Page 8, lines 1-2: This is interesting indeed.			Furthermore, some reflection on the adopted comparative case study research design with references would be desirable.	1	2	admsci5040213_devetyaro	0
+In: European Review of Industrial Economics and Policy (5).			However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between.	1	2	admsci5040213_devetyaro	0
+The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters.			However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified. The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between.	1	2	admsci5040213_devetyaro	0
+http://revel.unice.fr/eriep/index.html?id=3543, accessed March 13th, 2015.			Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment.	1	2	admsci5040213_devetyaro	0
+P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.			Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment.	1	2	admsci5040213_devetyaro	0
+For instance, what does it mean if a regional cluster organisation is pre-dating the national policy programme it is supposed to implement, as evident from some of the cases?			As also elaborated above, the conclusion should go beyond a mere summary to include methodological reflection, implications for policy (learning), as well as a more extensive discussion of further research perspectives.	1	2	admsci5040213_devetyaro	0
+KOSCHATZKY, K., 2000: A river is a river – Cross-border networking between Alsace and Baden.			Page 3, line 3: The authors might want to link to the literature on differentiated knowledge bases, which assigns an analytical (science-based) knowledge base to the biotech industry (cf. ASHEIM ET AL. 2011).	1	2	admsci5040213_devetyaro	0
+KOSCHATZKY, K., 2000: A river is a river – Cross-border networking between Alsace and Baden.			Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references. If ‘few’ means ‘no’, then please say so straightaway.	1	2	admsci5040213_devetyaro	0
+DURANTON, G.; MARTIN, P.; MAYER, T.; MAYNERIS, F., 2010: The economics of clusters: Lessons from the French experience.			Page 6, line 25: I would suggest softening the statement that matched funding ‘prevents moral hazard and crowding out’. It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.	1	2	admsci5040213_devetyaro	0
+DURANTON, G.; MARTIN, P.; MAYER, T.; MAYNERIS, F., 2010: The economics of clusters: Lessons from the French experience.			Section 2 contains a number of “expectations”, which could be flagged out and numbered as hypotheses. Taking these up in the empirical discussion could help the reader’s orientation.	1	2	admsci5040213_devetyaro	0
+This dichotomy should be refined and the classification of cases qualified as, e.g., ‘relatively bottom-up’.			Page 8, lines 1-2: This is interesting indeed. Is there comparable evidence on policy learning between Germany and France? Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.	1	2	admsci5040213_devetyaro	0
+This dichotomy should be refined and the classification of cases qualified as, e.g., ‘relatively bottom-up’.			Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.	1	2	admsci5040213_devetyaro	0
+Freeman, C., 1988: Japan: A new National System of Innovation?			Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.	1	2	admsci5040213_devetyaro	0
+http://revel.unice.fr/eriep/index.html?id=3495, accessed June 13, 2015.			Section 3.3: There is a little bit of literature on French national cluster policy that deserves a brief review or at least reference here (e.g., BRETTE/CHAPPOZ 2007, DURANTON ET AL. 2010, LONGHI/ROCHHIA 2013).	1	2	admsci5040213_devetyaro	0
+BRETTE, O.; CHAPPOZ, Y., 2007: The French competitiveness clusters: Toward a new public policy for innovation and research?			Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.	1	2	admsci5040213_devetyaro	0
+; BOSCHMA, R.; COOKE, P., 2011: Constructing regional advantage: Platform policies based on related variety and differentiated knowledge bases.			Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’? If the aim of public policy is to grow a cluster around a research organisation or hospital, this does not seem to produce any meaningful outcomes as yet.	1	2	admsci5040213_devetyaro	0
+Broad Comments • However, the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2.			Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’. This should be backed up with evidence (data) and preferably rephrased, such as ‘recognised research universities’, or ‘elite’ with reference to the German federal government’s programme of university excellence (Exzellenzinitiative).	1	2	admsci5040213_devetyaro	0
+Would suggest to use these case studies as a source of ideas on how policies and cluster initiative management might relate to each other; that would make this a more interesting paper.			Page 15, line 27: Repetition – the IZB incubator was already mentioned on line 9.	1	2	admsci5040213_devetyaro	0
+In: European Planning Studies, 8(4): 429-449.			Section 4.5: Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that KOSCHATZKY (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.	1	2	admsci5040213_devetyaro	0
+• The layout of the tables may be improved, especially the spacing of the text.			The layout of the tables may be improved, especially the spacing of the text.	1	2	admsci5040213_devetyaro	0
+In: Journal of Economic Issues, 41(2): 391-398.			The paper contains funding information in Yen and Euros. This should be harmonised, or preferably conversions given in brackets or footnotes for the reader’s convenience.	1	2	admsci5040213_devetyaro	0
+‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters.			I wish to congratulate the authors for their careful and thoughtful revision. All remarks have been considered, most have been fully incorporated and the paper has gained substantially from revision. As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing. Since this may be managed by the editorial office, I do not see the need for another review.	3	2	admsci5040213_devetyaro	0
+regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.			Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators. If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences. This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation. A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).	3	2	admsci5040213_devetyaro	0
+Methodology: The choice of clusters is generally well explained on pp.			Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique. I would still like to see a qualification stating that private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.	3	2	admsci5040213_devetyaro	0
+Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.	3	2	admsci5040213_devetyaro	0
+In: European Planning Studies, 8(4): 429-449.			P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before. Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.” In response, the authors now cite this paper in footnote 14 in Section 3.2. to support the view that BioValley has hardly had any impact in terms of intensifying cross-border interaction. This reformulation is incorrect as the Koschatzky (2000) paper pre-dated the BioValley initiative and does not look at INTERREG either. Unfortunately, my claim regarding the limited impact of BioValley on cross-border networking draws on conversations with practitioners and a field trip to the region, but cannot be supported by a reference as far as I know. I would therefore suggest to drop this footnote altogether.	3	2	admsci5040213_devetyaro	0
+Freeman, C., 1988: Japan: A new National System of Innovation?			P. 16: Sources should be stated underneath table 2.	3	2	admsci5040213_devetyaro	0
+A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).			P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.	3	2	admsci5040213_devetyaro	0
+If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences.			P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?	3	2	admsci5040213_devetyaro	0
+Author Response Author Response File: Author Response.docx			Why Jung? The authors have selected Jung’s archetypes as the theoretical foundation for the paper.	1	2	admsci6020005_makarova	0
+Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).			This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.	1	2	admsci6020005_makarova	0
+While Jungian approaches have been used by marketing and consumer researchers in the past (e.g., Hirschman), the reasons for this choice need to be justified in the context of other alternatives.			The abstract states: “Aaker is seen by many as the branding guru” and refers to Aaker (1997) in the introductory paragraph. Are you referring to David Aaker (the father) or Jennifer Aaker (the daughter)?	1	2	admsci6020005_makarova	0
+Personally, I agree with you – brand personality is a key facet of brand identity.			Indeed, Aaker (1997) is a highly-cited work. Are there any other limitations or criticisms the reader should be informed of? However, you may want to tell readers how brands can convey aspects of personality via advertising (see: Allen and Olson 1995).	1	2	admsci6020005_makarova	0
+We would have loved to substantiate the archetype concept by neurophysiological research but were not convinced about the reliability of claimed research findings so we added it under scope for further research (chapter: Implications).			Page 2, line 62: I disagree with the claim that “above models can be seen as extensions of Aaker’s (1997) brand personality model.” The models you listed are variants of well-known hierarchy of effects models.	1	2	admsci6020005_makarova	0
+"For example the fact that some ""heroes"" can move across categories - e.g., Tiger Woods for Buick."			Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition.	1	2	admsci6020005_makarova	0
+However, you may want to tell readers how brands can convey aspects of personality via advertising (see: Allen and Olson 1995).			The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.	1	2	admsci6020005_makarova	0
+We are delighted that our reviewer sees the link (as the authors do) between Aaker and Jung’s images of Freedom, Social, Order and Ego.			Evidence of claim? Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way. In contrast the 55 Asian students knew the chains.” Was brand familiarity actually measured, or was it assumed?	1	2	admsci6020005_makarova	0
+Methodology (a) Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition.			However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).	1	2	admsci6020005_makarova	0
+However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).			Clarity and elaboration of Page 5+: The methods section lacks clarity – it would be very difficult for a researcher to replicate the procedure. Please provide greater detail of the scales used, the anchor terms, and how the survey/rating task was administered (online? Paper and pencil?).	1	2	admsci6020005_makarova	0
+After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity.			“Archetype Articulate Charisma” (heading title in Table 3): This is not clear. Either define the term in the text or use phrasing that is clear to the reader.	1	2	admsci6020005_makarova	0
+Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes.			Findings: Who rated the three personality traits? The student/respondents? How many items were used? Why do you not report reliability measures for the items and scale?	1	2	admsci6020005_makarova	0
+It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego.			Factor analysis: Have you reported principal components, or a rotated solution?	1	2	admsci6020005_makarova	0
+by means of neurophysiological evidence based research but tried in vain.			Interpretation: The interpretation of Archetypes seems to assume that the types are mutually-exclusive. However, this is not really the case.	1	2	admsci6020005_makarova	0
+The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.			I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some.	1	2	admsci6020005_makarova	0
+We hope that we have given full consideration to all comments.			I think that the relevance of the research to practitioners as well as academics needs to include some current findings about ads, their placement and effectiveness.	1	2	admsci6020005_makarova	0
+The general and growing aversion to ads - ad blocker or the increased use of native advertising - sponsored branded content - that appears in desired media channels is a major issue for ad agencies.			Why Jung? The authors have selected Jung’s archetypes as the theoretical foundation for the paper. While Jungian approaches have been used by marketing and consumer researchers in the past (e.g., Hirschman), the reasons for this choice need to be justified in the context of other alternatives. Jungian approaches suffer from two core limitations: (a) the assertion of universality – which the authors note, and (b) the psychoanalytic heritage of the ideas (which presupposes a certain view of motivation) is disavowed by most contemporary psychologists.	1	2	admsci6020005_perova	0
+Author Response Author Response File: Author Response.docx			Literary archetypes: An alternative approach to studying the archetypal aspects of brand image can be found by scholars who adopt a literary or cultural view of archetypes, such as the ones advanced by Joseph Campbell or Northrop Frye (see, for example: Stern 1989). One could make the argument that using archetypes in advertising has greater affinities to mythology, literature and communications. Work on narrative theory and characterization in advertising also aligns with the archetype approach (see, for example: Mulvey and Medina 2003; Padgett and Allen 1997; Scott 1994; Stern 1988). This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.	1	2	admsci6020005_perova	0
+Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way.			The abstract states: “Aaker is seen by many as the branding guru” and refers to Aaker (1997) in the introductory paragraph. Are you referring to David Aaker (the father) or Jennifer Aaker (the daughter)?	1	2	admsci6020005_perova	0
+We found them very useful and they increased the value of our paper considerably.			Indeed, Aaker (1997) is a highly-cited work. Are there any other limitations or criticisms the reader should be informed of? There has been some backlash to the application of brand personality to inanimate objects. This may be of concern, because the paper uses hotels as an example. Personally, I agree with you – brand personality is a key facet of brand identity. However, you may want to tell readers how brands can convey aspects of personality via advertising (see: Allen and Olson 1995). This is highly relevant to answering the “so what?” question that is addressed in the implications section.	1	2	admsci6020005_perova	0
+Jungian approaches suffer from two core limitations: (a) the assertion of universality – which the authors note, and (b) the psychoanalytic heritage of the ideas (which presupposes a certain view of motivation) is disavowed by most contemporary psychologists.			Page 2, line 62: I disagree with the claim that “above models can be seen as extensions of Aaker’s (1997) brand personality model.” The models you listed are variants of well-known hierarchy of effects models. Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).	1	2	admsci6020005_perova	0
+(b) Sample size: A critical view would state that the present research relies on a very small sample of advertisements (n=2) to make its claims.			Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition. Grounded theory is based on the idea of immersion in data to induct new conceptual categories. I see the present study as using existing typologies and categories – there are no new concepts that emerge from the research that can be applied directly to future research.	1	2	admsci6020005_perova	0
+The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.			Sample size: A critical view would state that the present research relies on a very small sample of advertisements (n=2) to make its claims. After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity. The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression. Though such studies are rare, good interpretive and quantitative examples can be found in the literature. At a minimum, this exploratory paper should set a more specific and ambitious plan for systematic future research on the topic. For example, Padgett and Mulvey (2007) illustrate a method to identify personal values conveyed by 16 ad campaigns (another complementary dimension of personified brands) and Padgett and Mulvey (2009) demonstrate a novel way to characterize customer-brand relationship archetypes. Imagine a conversation with these authors: what do you contribute to the ongoing conversation?	1	2	admsci6020005_perova	0
+One could make the argument that using archetypes in advertising has greater affinities to mythology, literature and communications.			Evidence of claim? Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way. In contrast the 55 Asian students knew the chains.” Was brand familiarity actually measured, or was it assumed? The claim is very absolute – I wonder if any of them ever travelled internationally – presumably EMBAs would be more worldly or cosmopolitan than this… (d)	1	2	admsci6020005_perova	0
+I hope my suggestions are given full consideration, as I believe that they will assist them in achieving this important objective.			Figure 3: Because you are studying commercials (which unfold over time), I don’t think the single screenshots really add much value to the paper. However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).	1	2	admsci6020005_perova	0
+There has been some backlash to the application of brand personality to inanimate objects.			Clarity and elaboration of Page 5+: The methods section lacks clarity – it would be very difficult for a researcher to replicate the procedure. Please provide greater detail of the scales used, the anchor terms, and how the survey/rating task was administered (online? Paper and pencil?).	1	2	admsci6020005_perova	0
+There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.			“Archetype Articulate Charisma” (heading title in Table 3): This is not clear. Either define the term in the text or use phrasing that is clear to the reader. Please don’t expect them to consult original sources by Jung!	1	2	admsci6020005_perova	0
+The claim is very absolute – I wonder if any of them ever travelled internationally – presumably EMBAs would be more worldly or cosmopolitan than this…			Findings: Who rated the three personality traits? The student/respondents? How many items were used? Why do you not report reliability measures for the items and scale?	1	2	admsci6020005_perova	0
+Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes.			Factor analysis: Have you reported principal components, or a rotated solution? More details are required.	1	2	admsci6020005_perova	0
+Personally, I agree with you – brand personality is a key facet of brand identity.			Interpretation: The interpretation of Archetypes seems to assume that the types are mutually-exclusive. However, this is not really the case. A close reading of some of the cited work (i.e., Mark and Pearson 1991) recognizes that archetypes can be blended.	1	2	admsci6020005_perova	0
+"Scott, Linda M. (1994), ""The Bridge from Text to Mind: Adapting Reader-Response Theory to Consumer Research,"" Journal of Consumer Research, 21 (3), 461-80."			I hope my suggestions are given full consideration, as I believe that they will assist them in achieving this important objective.	1	2	admsci6020005_perova	0
+Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes.			I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some. Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes. It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego. There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.	1	2	admsci6020005_perova	0
+(h) Factor analysis: Have you reported principal components, or a rotated solution?			I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some. Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes. It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego. There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.	1	2	admsci6020005_perova	0
+The reviewer is completely right in adressing this point.			In the introduction, (Line 76 79), you present the objectives, but those are not clear, I respectfully suggest focusing on your hypothesis 2.	1	2	agriculture12060868_makarova	0
+The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			Line 78: You wrote, “designed to meet the basic behavioral needs of the sow at farrowing”, but this is not tested on the paper.	1	2	agriculture12060868_makarova	0
+The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			Lines 80 – 83 moves to discussion section 4.	1	2	agriculture12060868_makarova	0
+The system we have used is to refer to the table with statistical values when reporting significant differences for the results presented in figures and tables, but for those that are presented in the text only, we also have P- and F-values in brackets.			Figure 1 is missing 5.	1	2	agriculture12060868_makarova	0
+The reviewer is completely right in adressing this point.			Figure 2 needs more description, maybe include which wall… Please describe units of the dimensions 6.	1	2	agriculture12060868_makarova	0
+There are only minor comments with very specific issues:			Figure 4. Please indicate which is “sowconf” and which is “trad”. In the left graph the units are in m2, but in right side it does not have unit, seems ar?	1	2	agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			Line 206: I don’t understand why you use just 10 o 12 sows, 50% primi and multiparous, but in line 225 mentioned that “there were no secure information about sow parity” please be consistent.	1	2	agriculture12060868_makarova	0
+The reviewer is completely right in adressing this point.			Please include P values when report significant differences.	1	2	agriculture12060868_makarova	0
+The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			Figure 1 does not appear in the pdf that this reviewer could download. Only the Figure Caption appears.	1	2	agriculture12060868_makarova	0
+The following changes could improve the quality of the paper.			It is not totally clear to the reviewer how stillborn piglets were evaluated as compared to “piglets with no milk in stomach”.	1	2	agriculture12060868_makarova	0
+There are only minor comments with very specific issues:			Line 224-225, it states that that no information/security on the productivity data on whether it was primiparous or multiparous sow. Again this creates a bit of confusion. Sows were recorded, 50% primiparous and 50% multiparous to evaluate behaviour, and results are discussed accorging to that (lines 320-321). Why productivity data was not available?	1	2	agriculture12060868_makarova	0
+However, it would be more elegant to have more batches.			Figure 5 and 7 display the overall causes of mortality for the three batches. However, differences in batches were found. Could this overall causes be biased by this batch effect?	1	2	agriculture12060868_makarova	0
+The writing and data presenting need to be improved.			Please add the important information to the footnote or figure legend of the tables and figures. Such as: abbreviations, replicates n=?, mean±SD/SE, ect.	1	2	agriculture12060868_makarova	0
+As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			Please italic the P value throughout the paper.	1	2	agriculture12060868_makarova	0
+Finally, thank you for being so patient with our manuscript			Lines 129-133, where is the figure 1?	1	2	agriculture12060868_makarova	0
+The following changes could improve the quality of the paper.			Line 263, please add “(3 batches with TRAD and 2 batches with SOWCOMF)” to the figure legends.	1	2	agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			Line 275-276, Space needed before and after “±”.	1	2	agriculture12060868_makarova	0
+Finally, thank you for being so patient with our manuscript			The quality of the Figrue 6 need to be improved.	1	2	agriculture12060868_makarova	0
+The writing and data presenting need to be improved.			Figure 7, using “*” to substitute the different letters to express the difference.	1	2	agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			Table 4, please using the writing way to present the table.	1	2	agriculture12060868_makarova	0
+The writing and data presenting need to be improved.			Figure8, do not understand the meaning of “***” here?	1	2	agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			Figure 7, please improve the perspective of the figure.	1	2	agriculture12060868_makarova	0
+The results are described and discussed in detail.			lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?	1	2	agronomy12061433_makarova	0
+Additional corrections through the text have been done.			How many seeds were collected?	1	2	agronomy12061433_makarova	0
+Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			line 115: Did one repetition mean one plant?	1	2	agronomy12061433_makarova	0
+Conclusion: What kind of broader impact of this work could be?			line 130: How many fruits were used as a sample of representative fruits?	1	2	agronomy12061433_makarova	0
+1) The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.			The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.	1	2	agronomy12061433_makarova	0
+We also thank you for the positive and encouraging comment.			In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green? Any explanation?	1	2	agronomy12061433_makarova	0
+2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			“products”	1	2	agronomy12061433_makarova	0
+We also thank you for the positive and encouraging comment.			Line 245: would be better to show the value of each agronomic trait mentioned here	1	2	agronomy12061433_makarova	0
+2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			Line 385-388: the figure legend does not provide enough information on this figure	1	2	agronomy12061433_makarova	0
+#Answer: Thanks We hope that these revisions successfully address the raised concerns, remaining available for any other requests regarding the manuscript.			Line 435: it may not be worth to mention the sweet pepper as the first 30 crops Line 468: “GBS”?	1	2	agronomy12061433_makarova	0
+line 135: How many fruits were used as a bulk of representative fruits?			Line 468: “GBS”? Genotype by sequencing?	1	2	agronomy12061433_makarova	0
+Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			What kind of broader impact of this work could be?	1	2	agronomy12061433_makarova	0
+Additional corrections through the text have been done.			lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?	1	2	agronomy12061433_perova	0
+Below point by point response to concerns Reviewer 1 The manuscript is generally well written.			How many seeds were collected?	1	2	agronomy12061433_perova	0
+We also thank you for the positive and encouraging comment.			line 115: Did one repetition mean one plant?	1	2	agronomy12061433_perova	0
+Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			line 135: How many fruits were used as a bulk of representative fruits?	1	2	agronomy12061433_perova	0
+This work is quite interesting and is well-written.			Conclusion: What kind of broader impact of this work could be?	1	2	agronomy12061433_perova	0
+Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.	1	2	agronomy12061433_perova	0
+Changes in respect to the previous version are in track changes.			In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green? Any explanation?	1	2	agronomy12061433_perova	0
+line 135: How many fruits were used as a bulk of representative fruits?			Line 37: “products”	1	2	agronomy12061433_perova	0
+See the attachment Comments for author File: Comments.pdf			Line 245: would be better to show the value of each agronomic trait mentioned here	1	2	agronomy12061433_perova	0
+We have carefully considered comments and thoughtful suggestions, revising the manuscript accordingly.			Line 385-388: the figure legend does not provide enough information on this figure	1	2	agronomy12061433_perova	0
+Additional corrections through the text have been done.			Line 435: it may not be worth to mention the sweet pepper as the first 30 crops	1	2	agronomy12061433_perova	0
+#Answer: Again, we would like to thank You for your attention and for carefully reading the manuscript.			Line 468: “GBS”? Genotype by sequencing?	1	2	agronomy12061433_perova	0
+Others: Line 37: “products” Line 245: would be better to show the value of each agronomic trait mentioned here Line 385-388: the figure legend does not provide enough information on this figure Line 435: it may not be worth to mention the sweet pepper as the first 30 crops Line 468: “GBS”?			Conclusion: What kind of broader impact of this work could be?	1	2	agronomy12061433_perova	0
+It is perfectly feasible that some owners did not leave their homes because they were for example, ill, but they did so during the pandemic.			A catchy topic, but its content is unoriginal, bringing nothing new to the pre-pandemic science. It seems that the authors of this study have already tackled this topic exhaustively in their previous articles	1	2	ani12040482_perova	0
+According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			The entire first paragraph of Discussion repeats what has already been written in earlier chapters (line 372-379).	1	2	ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			"Line 382-384 „To our knowledge, this is the first empirical evidence to show a link between changes in dogs’ leaving routines and risk of SRBs.” The authors take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed. And changing the routine of the day is a well-known factor influencing the dog's welfare and behavior, including the occurrence of separation anxiety, so the above-quoted sentence should be deleted because it is untrue. The authors have hardly studied all the publications on how dogs behave when daily routines were changed, so the term ""first study"" is exaggerated."	1	2	ani12040482_perova	0
+Larger issues: I would like to see the authors address a few different angles in the discussion 1.			Line 444 „This study has provided a unique and valuable insight into…..” the authors also mark their only unique role in the study, which does not bring new content.	1	2	ani12040482_perova	0
+The sample size is good, and the results are interesting.			My only suggestion is that making it clear why February and October were chosen as survey-points may be beneficial. This is discussed later on but highlighting here would be useful.	1	2	ani12040482_perova	0
+My only suggestion is that making it clear why February and October were chosen as survey-points may be beneficial.			Aa little more mention of issues of SRBs from owner perspective (e.g. noise complaints and issues with housing; property damage; decline of human-pet bond) could be beneficial (line 56-57).	1	2	ani12040482_perova	0
+We have addressed your comments and responded in the attached file.			It may be helpful to discuss why there is poor owner adherence to programmes to reduce SRB (Line 66-67).	1	2	ani12040482_perova	0
+The article is generally well-written, the topic is timely, important, and interesting from a dog welfare aspect.			I wasn't sure if line 79-80 was needed- about dogs having returned to pre-pandemic pattern, I think it is a little dismissive of your own research which is valuable and applicable beyond the pandemic.	1	2	ani12040482_perova	0
+Introduction: This is clear and in general provides a good background to the topic (bar a couple of points where more expansion would be beneficial).			Line 96-97- some more detail on the findings of these various surveys would be beneficial similarly to Bowen's study, just so provide a clear background/discussion of literature setting scene for your study.	1	2	ani12040482_perova	0
+The study reports that pet dogs whose leaving routines changed most during the COVID-19 pandemic (i.e., their owners left them alone less frequently) would be at greater risk of developing separation-related behaviours following the lifting of pandemic restrictions.			It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g. justification for this/why this time duration used.	1	2	ani12040482_perova	0
+Line 382-384 „To our knowledge, this is the first empirical evidence to show a link between changes in dogs’ leaving routines and risk of SRBs.” The authors take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed.			Line 131: There was an error with referencing wanted to flag.	1	2	ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			It may be useful to spell out a little more here why these survey periods were chosen here, this is covered elsewhere but think could be made clearer here and earlier.	1	2	ani12040482_perova	0
+The authors still take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed.			Figure 1- Perhaps just double-check this is referred to in the text. Think either way this figure would benefit a bit of context on why was included/what was showing in context of this study as currently info is a bit generic and didn't totally tie into people's working from home etc.	1	2	ani12040482_perova	0
+They do not seem to state dog is favouring them less unless text was omitted here.			However, I don't think the figures were referred to in the text (unless that was the errors highlighted below?)	1	2	ani12040482_perova	0
+Lines 383-384: Change “dogs’ leaving routines” to “dogs’ time left alone” Thank you, this edit has been applied.			There were some errors where reference source not found which would want to look into (line 246; 265; 274; 292; 305; 323 etc).	1	2	ani12040482_perova	0
+Lines 383-384: Change “dogs’ leaving routines” to “dogs’ time left alone” Thank you, this edit has been applied.			Line 366-369= I'm not sure it's totally fair to interpret this (the dog not always bringing toy when greeting them) as an anthropomorphic interpretation on the part of the owner- the anthropomorphic interpretation seems to be from authors of manuscript not owner? They do not seem to state dog is favouring them less unless text was omitted here.	1	2	ani12040482_perova	0
+L238 Do the authors have information about the Owner demographics?			I felt the discussion about changes in greeting behaviour as an early indicator was interesting and could be expanded. Some more detail on the application of the findings to practitioners/owners would be beneficial. I do think this is of interest to a range of stakeholders.	1	2	ani12040482_perova	0
+Manuscript has been slightly improved, but my comments were not included.			P6 L176 I am assuming you excluded dogs that were not left alone in February (or were there none?)	1	2	ani12040482_perova	0
+Line 366-369= I'm not sure it's totally fair to interpret this (the dog not always bringing toy when greeting them) as an anthropomorphic interpretation on the part of the owner- the anthropomorphic interpretation seems to be from authors of manuscript not owner?			P10 L307-308 Could you make this a positive statement to show highlight the impact of amount of change on SRB: Dogs whose days left alone per week increased the most…If the data support that angle, I think it’s a stronger statement than what is written.	1	2	ani12040482_perova	0
+Larger issues: I would like to see the authors address a few different angles in the discussion 1.			1. That dogs showing SRB prior to lockdown were more likely to show it after. Suggests the behavior is fairly robust and dogs having shown it at one point continue to be at risk (without knowing what interventions were taken)	1	2	ani12040482_perova	0
+In other words, 1407/1807 (78%) of dogs were clear of SRB in February and 90.1% in October.			2. Can you address the dogs that improved between Feb and Oct? You report the number of new dogs showing SRB but can you report on dogs that went the other way? What was the actual # and %? It seems you have compared within dogs that showed SRB in baseline and other models for dogs that did not. Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing. This comparison seems essential—if some dogs get worse but some get better, than the argument that changes due to COVID might bring about new SRB becomes weaker.	1	2	ani12040482_perova	0
+We have addressed your comments and responded in the attached file.			3.60 One of the challenges in this is we don’t know background rates of change for dogs with SRB. That is, would the same % of dogs changed (in either direction) regardless of changes in owner schedules? We might see some dogs become sensitized over time and develop SRB (or something else changes in their life to precipitate it), while others might habituate or their behavior undergo extinction. As such, we can’t say for certain that these changes are due to changes in owner schedule. You note that the pseudo-R2 was small but I think you could go into more details and note the possibility that it was unrelated too. However, your results that the greater the change → the greater the new risk of SRB does lend support to the idea that this change in management might be a factor. I would suggest highlighting and discussing this outcome more.	1	2	ani12040482_perova	0
+L382 Please refer to Flannigan and Dodman (2001) here (owner’s work schedule affects separation problems).			Title: change “Leaving Pattern” to “Time Spent Away”; leaving pattern is less clear (it could be frequency of leaving each day)	1	2	ani12040482_perova	0
+It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g justification for this/why this time duration used.			Line 214: if possible (might not be possible if this was how it was phrased in the survey), change “different to before” to “different from before”	1	2	ani12040482_perova	0
+This is good news, and it is unclear for me why is it reported as bad news.			Lines 383-384: Change “dogs’ leaving routines” to “dogs’ time left alone”	1	2	ani12040482_perova	0
+An error message occurred in several places (e.g., line 131, line 246, line 265)			Line 151: how was the criterion of “at least 5 minutes” chosen?	1	2	ani12040482_perova	0
+However, the authors do not focus on this result and do not explain this finding.			My main question when reading this section concerned how owners would know that certain separation-related behaviors occurred when they were away? There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing? This topic is not addressed until the Discussion section (line 395) but should be addressed here in the Methods and Materials section. Were behaviors, including different vocalizations, defined for owners? For example, what is the difference between pined, whined, and cried?	1	2	ani12040482_perova	0
+My main question when reading this section concerned how owners would know that certain separation-related behaviors occurred when they were away?			Also, I might have missed it, but did all of these owners live alone with their dog(s), meaning no other people in the household? If other people were present, did all of them have to be away or just the dog’s owner? This information might be presented in your other paper, but it would be good to include this basic information here as well, so readers can better understand your methods and findings.	1	2	ani12040482_perova	0
+L22 and elsewhere: dogs’ “leaving time” or “leaving hours” I think this expression is misleading because it is not the dogs that have left home, but the owners.			Line 166: define “devolved nations” for readers unfamiliar with the term.	1	2	ani12040482_perova	0
+Impact of the first COVID-19 lockdown on management of pet dogs in the UK.			Figure 2: should the blank box for “Feb, Not at all” be 0.0%?	1	2	ani12040482_perova	0
+Line 96-97- some more detail on the findings of these various surveys would be beneficial similarly to Bowen's study, just so provide a clear background/discussion of literature setting scene for your study.			Figure 4 is not called out in the text of the Results section.	1	2	ani12040482_perova	0
+The study reports that pet dogs whose leaving routines changed most during the COVID-19 pandemic (i.e., their owners left them alone less frequently) would be at greater risk of developing separation-related behaviours following the lifting of pandemic restrictions.			I believe Animals requires a Conclusions section.	1	2	ani12040482_perova	0
+The whole section is not relevant to the main question.			Line 100: insert “the” after “in”	1	2	ani12040482_perova	0
+According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			Line 355: delete the first “the”	1	2	ani12040482_perova	0
+An error message occurred in several places (e.g., line 131, line 246, line 265)			However, currently, the paper seems too long, and it goes into too much detail about how the pandemic took place in the UK. This information would have been interesting if data on the mood, stress level, general mental health of the owners had also been collected in connection with the pandemic and the links with dog behaviour have been investigated. But the questionnaire only asked how long the dogs had been alone and what they were doing during this time. Therefore, the detailed description of the pandemic is unnecessary and could be moved to the Supplemental material, together with Figure 1.	1	2	ani12040482_perova	0
+"The authors have hardly studied all the publications on how dogs behave when daily routines were changed, so the term ""first study"" is exaggerated."			I think that according to the statistics, the strongest finding is that dogs who showed SRB in February or May had increased odds of having SRB during the subsequent measures which is not surprising. What is surprising though is that 55.7% of the dogs who were reported to show SRB in February (before the pandemic) were clear in October which is good news. However, the authors do not focus on this result and do not explain this finding.	1	2	ani12040482_perova	0
+For example, what is the difference between pined, whined, and cried?			L18-20 „Whether dogs showed SRBs or not changed considerably over the months of the study, and one in ten dogs were found to have developed new SRBs in October, that they didn’t show before the pandemic” Overall, the percentage of dogs showing SRBs decreased during the study, and this should be clarified in the sentence above. What I see in Figure 4 and in the text is that approx. half of the dogs from the SRB group moved to the Clear group in October.	1	2	ani12040482_perova	0
+We have addressed your comments and responded in the attached file.			L22 and elsewhere: dogs’ “leaving time” or “leaving hours” I think this expression is misleading because it is not the dogs that have left home, but the owners. But this is the opinion of a person using English as a second language.	1	2	ani12040482_perova	0
+What was the proportion of owners who did not leave their dog at home before the pandemic?			L45 Separation behaviour, separation-related behaviour, separation distress, separation anxiety terms are used interchangeably throughout the text, although these terms do not necessarily describe the same behaviours. Please provide a clear definition of the behaviour aimed to study and stick to one term.	1	2	ani12040482_perova	0
+Study periods: It may be useful to spell out a little more here why these survey periods were chosen here, this is covered elsewhere but think could be made clearer here and earlier.			L92 delete the comma after Bowen et al.’s.	1	2	ani12040482_perova	0
+What was the proportion of owners who indicated that they would like to participate in the follow-up study?			L92-92 28.5% of dogs reported to have SRPs – this is a much higher number than mentioned in previous studies and needs to be discussed.	1	2	ani12040482_perova	0
+If other people were present, did all of them have to be away or just the dog’s owner?			L118 Materials and Methods This section (after the Ethics) should begin with the Subjects section. It is confusing to read about the surveys without knowing the sample sizes per survey and the demographics.	1	2	ani12040482_perova	0
+This topic is not addressed until the Discussion section (line 395) but should be addressed here in the Methods and Materials section.			L127-137 The dates of data collections have been already mentioned in L121-126, so there is no need for repeating them. The whole section is not relevant to the main question.	1	2	ani12040482_perova	0
+What was the proportion of owners who indicated that they would like to participate in the follow-up study?			L131-132 and elsewhere: (Error! Reference source not found.) This error occurs 7 times in the text, please correct.	1	2	ani12040482_perova	0
+Conclusions: I believe Animals requires a Conclusions section.			L148 Please explain why a period of 7 days were asked to be reported	1	2	ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			L165-167 Figure 1. This Figure should be moved to the Supplement material as it tells nothing about the dogs’ behaviour.	1	2	ani12040482_perova	0
+Line 29: change “dogs whose leaving hours reduce most” to “dogs whose time left alone decreased most” Thank you, this edit has been applied.			L216-218 “The text was then coded by two researchers” I could not find agreement data reported later.	1	2	ani12040482_perova	0
+P10 L307-308 Could you make this a positive statement to show highlight the impact of amount of change on SRB: Dogs whose days left alone per week increased the most…If the data support that angle, I think it’s a stronger statement than what is written.			L238 Do the authors have information about the Owner demographics? What was the proportion of females? Was there a relationship between age and leaving hours? Or explain why this information has not been used.	1	2	ani12040482_perova	0
+As such, we can’t say for certain that these changes are due to changes in owner schedule.			L232-242 These sections should be moved to the Materials and Methods as a “Subjects” section	1	2	ani12040482_perova	0
+Impact of the first COVID-19 lockdown on management of pet dogs in the UK.			L232 How many owners have filled in the questionnaire altogether? What was the proportion of owners who did not leave their dog at home before the pandemic? Why were they excluded? It is perfectly feasible that some owners did not leave their homes because they were for example, ill, but they did so during the pandemic. What was the proportion of owners who indicated that they would like to participate in the follow-up study? Among them, what was the proportion of owners who has not responded to the request afterward?	1	2	ani12040482_perova	0
+Figure 4 is not called out in the text of the Results section.			L278-280 “There were 1,407 dogs who were clear of SRB at baseline in February 2020. When looking at SRB status in October, of 1,187 dogs who were left alone in October, 117 (9.9%) were reported to have shown at least one SRB”. In other words, 1407/1807 (78%) of dogs were clear of SRB in February and 90.1% in October. This is good news, and it is unclear for me why is it reported as bad news. The next section (l281-283) emphasizes that 55.7% of the dogs who were reported to show SRB in February were clear in October. This is a big improvement, I think.	1	2	ani12040482_perova	0
+L216-218 “The text was then coded by two researchers” I could not find agreement data reported later.			L285 Figure 4. This is a great figure, thank you for including it. However, an explanation would be useful for those readers who are not familiar with this type of depicting.	1	2	ani12040482_perova	0
+However, I don't think the figures were referred to in the text (unless that was the errors highlighted below?)			L291 The odds ratio (OR) is 5.38 and not 4.38 in Table 1.	1	2	ani12040482_perova	0
+L148 Please explain why a periof of 7 days were asked to be reported			L293-295 “The final variable associated with October SRB status was the difference in the number of days dogs were left alone for between the February baseline and lockdown”.	1	2	ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			OR was 0.81 here. Please explain the relationship between the variables.	1	2	ani12040482_perova	0
+The authors examined how separation-related behaviors in pet dogs changed in relation to time spent away from home by UK owners in February 2020 (pre-pandemic), May-July 2020 (first lockdown during the pandemic), and October-November 2020 (local tier-system for restrictions; follow-up).			L298 Table 1 legend: Add what bold indicates L307 The odds ratio (OR) is 4.97 in Table 1 and not	1	2	ani12040482_perova	0
+This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			L307 The odds ratio (OR) is 4.97 in Table 1 and not 3.60 Thank you!	1	2	ani12040482_perova	0
+Larger issues: I would like to see the authors address a few different angles in the discussion 1.			L314 Table 2: bold is not used here although it was used in Table 1	1	2	ani12040482_perova	0
+"The authors have hardly studied all the publications on how dogs behave when daily routines were changed, so the term ""first study"" is exaggerated."			L333 table 3: same: bold is not used here	1	2	ani12040482_perova	0
+According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			L382 Please refer to Flannigan and Dodman (2001) here (owner’s work schedule affects separation problems). Flannigan, G., & Dodman, N. H. (2001). Risk factors and behaviors associated with separation anxiety in dogs. Journal of the American Veterinary Medical Association, 219(4), 460-466.	1	2	ani12040482_perova	0
+Results: These were generally well-detailed with clear layout considering scope of results.			L395-397 “This value is likely to be an underestimate” The underestimation has already occurred during the baseline.	1	2	ani12040482_perova	0
+Authors addressed all my requests and suggestions to my fullest satisfaction.			Figure 7b is not very credible, when U937 become macrogphages adhere to the plate and assume the morphology of a macrophage. the figure is not clear and not well visible.	1	2	antiox11040683_makarova	0
+Future work could extend our analysis and examine the antioxidant capacity of the BCAT1 CXXC motif in other FAB subtype models, for example HL60 (AML, M2) and NB4 (AML M3 aka APL) [45].”			The authors should also show the flow cytometers panels related to cd68 and cd14.	1	2	antiox11040683_makarova	0
+Because of their pro-monocytic phenotype, many regard the U937 cell line to be representative of an AML subtype M5, under the French-American-British (FAB) classification system [43–46].			Small comment, the supplementary Fig. S4B would need some compensation work....this would be better to correct before final approval for publication ( up to editor).	1	2	antiox11040683_makarova	0
+We agree with your comments and have addressed these to the best of our ability in the revised manuscript.			Also the gating strategy on Fig. 4d ( is going in right direction), what I am missing here is the exclusion of dead cells?).	1	2	antiox11040683_makarova	0
+The Authors showed the role of the BCAT1 CXXC motif in AML and to understand how this may influence cellular redox processes.			The data presented by the authors are robust and interesting but are based on a single AML cell line, which is not a bonafide AML cell line (derived from the pleural effusion of a 37-year-old, White, male patient with histiocytic lymphoma, according to ATCC).	1	2	antiox11040683_makarova	0
+"At the beginning of the last paragraph of Introduction: ""In this work, based on the design concept of improving the tensile stiffness of LET joints in Ref."			Transient analysis followed by reliability experiments/analysis/modeling of the proposed structure would be interesting and critical in aspect of joint application contrains. In my opoinion is should be added or explained in context of scheduled follow-up activities.	1	2	app12052642_perova	0
+Chen, G.; Ma, F.; Hao, G.; Zhu, W. Modeling large deflections of initially curved beams in compliant mechanisms using 277 chained Beam-Constraint-Model, ASME J. Mech.			The article being evaluated is an extension of the above cited reference, and the authors would be well advised as to summarize this article, in part, and to delineate what has changed as of 2019, i.e. put this in a paragraph and to explain the subsequent evolution since 2019.	1	2	app12052642_perova	0
+Author Response We thanks the reviewer for the encouragement and valuable comments.			Secondly what is missing in the article is in terms of application a clear delineation as to what benefits robotics accrue by the developments summarized in this article . What new capacities and abilities so created by this evolutionary jump Aside from that the article is solidly researched and is useful as a source of technology development	1	2	app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			Keywords are not sufficient after the abstract.	1	2	app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			Work relevant to the research topic is not incorporated, i.e., the work done by other researchers / authors in this field are not incorporated.	1	2	app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			References are not written in the standard journal format.	1	2	app12052642_perova	0
+10 + description) steady state inverstigation is acceptable.			Work relevant to the research topic is not incorporated, i.e., the work done by other researchers / authors in this field are not incorporated.	1	2	app12052642_perova	0
+The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			Please add more details regarding paper’s novelty, it is not very clear what are the novelties of this paper.	1	2	app12052642_perova	0
+The current paper proposes a novel lamina emergent torsional (LET) joint named double-laminated lamina emergent torsional joint (DL-LET) joint which utilizes a layer of flexible H18 aluminum foil to increase the tensile stiffness of LET joint.			Please add more details of how the theory from the first sections is applied in the results section and better detail the theoretical part, in this version the theoretical part is almost inexistent.	1	2	app12052642_perova	0
+[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			Please detail how the parameters were obtained.	1	2	app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			The state of the art it is very poor regarding representative papers, maybe the author could add the following publications: o Hybrid Data-Driven Fuzzy Active Disturbance Rejection Control for Tower Crane Systems, European Journal of Control, vol. 58, pp. 373-387-11, 2021. o Event-Triggered Adaptive Fuzzy Control for Stochastic Nonlinear Systems with Unmeasured States and Unknown Backlash-Like Hysteresis, IEEE Transactions on Fuzzy Systems, doi 10.1109/TFUZZ.2020.2973950, pp. 1–19, 2020.	1	2	app12052642_perova	0
+The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			Please add more details regarding the obtained results.	1	2	app12052642_perova	0
+Nice piece of research but one aspect needs to be addressed.			Add the both the advantages and the disadvantages of the proposed method. In the proposed manuscript only the advantages are presented.	1	2	app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			"We agree that transient analysis is important in designing a robot joint. However, the transient analysis requires relevant parameters of the robot arm be determined besides the parameters of the joint. The following sentence has been added at the end of Conclusions: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors"""	2	1	app12052642_perova	0
+Author Response We thanks the reviewer for the encouragement and valuable comments.			The following sentence has been added at the end of first paragraph of Conclusions to clarify disadvantage of DL-LET: “The DL-LET joint is vulnerable to delamination of the two layers.”	2	1	app12052642_perova	0
+Following the referee’s suggestion we have added the reference to Mekhov’s paper.			Other examples of many-atom-cavity continuous observation papers: W. Niedenzu, S. Schütz, H. Habibian, G. Morigi, and H. Ritsch, Phys. Rev.  88, 033830 (2013). I. B. Mekhov and H. Ritsch, Phys. Rev. Lett. 102, 020403 (2009). J. F. Corney and G. J. Milburn, Phys. Rev. A 58, 2399 (1998). I. D. Leroux, M. H. Schleier-Smith, and V. Vuletić, Phys. Rev. Lett. 104, 073602 (2010).	1	2	atoms4010002_makarova	0
+However, a change in the depth of a periodic potential does not change the Bloch frequency because this depends on the lattice’s spatial period, not the depth.			The role of sensing may in the end be less important and could be downplayed a bit in the manuscript.	1	2	atoms4010002_makarova	0
+However, a change in the depth of a periodic potential does not change the Bloch frequency because this depends on the lattice’s spatial period, not the depth.			What is the role of excitations to higher Bloch bands?	1	2	atoms4010002_makarova	0
+To be clear, other motional frequencies are altered: because the intracavity lattice depth is modulated by the backaction this will affect certain types of atomic motion, for example the oscillation frequency of an atom about the bottom of one of the potential minima \cite{Horak00}.			(1) It should be somehow mentioned more clearly (maybe in the abstract) that the idea is not completely new but work presents a new and more intutive picture of the system dynamics.	1	2	atoms4010002_makarova	0
+Hence, in principle I consider it suitable for publication.			(2) It is very helpful to connect cavity output spectra and forces. To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40. This should be mentioned.	1	2	atoms4010002_makarova	0
+Our work is very much still in progress and this paper will not, we hope, be our final word on this subject.			(3) The authors claim there should be no backaction of the field on the oscillation frequencies. This is somehow in contradiction with the eigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.	1	2	atoms4010002_makarova	0
+This change in potential depth due to backaction is fully included in our treatment.			Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.	1	2	atoms4010002_makarova	0
+This is because the presence of atoms in the cavity shifts the cavity resonance and hence the depth of the intracavity lattice: the Bloch energies are dependent on the potential depth.			(a) any measurement of the output field will generate backaction onto the condensate (Mekhov, J. Phys. B: Atomic, Molecular and Optical Physics, 45(10), 102001). This will generate extra noise, but might also help as it reduces atom number uncertainty in the cavity.	1	2	atoms4010002_makarova	0
+This will generate extra noise, but might also help as it reduces atom number uncertainty in the cavity.			(b) There will be heating limiting the available measurement time.	1	2	atoms4010002_makarova	0
+Rev. A 88, 033830 (2013). I. B. Mekhov and H. Ritsch, Phys. Rev. Lett. 102, 020403 (2009). J. F. Corney and G. J. Milburn, Phys. Rev. A 58, 2399 (1998). I. D. Leroux, M. H. Schleier-Smith, and V. Vuletić, Phys. Rev. Lett. 104, 073602 (2010).			There are other papers that link BECs, sensing and continuous quantum measurement process (in the absence of a cavity) M. Saba, T. A. Pasquini, C. Sanner, Y. Shin, W. Ketterle, and D. E. Pritchard, Science 307, 1945 (2005).	1	2	atoms4010002_perova	0
+To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40.			The link to the sensing of weak forces (in this case gravity) is interesting, but I am not convinced whether this is a practical approach. The estimated accuracy is $10^6$ that does not sound great.	1	2	atoms4010002_perova	0
+Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.			What is the role of excitations to higher Bloch bands?	1	2	atoms4010002_perova	0
+Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.			Have the effects of radiation pressure forces, etc., taken into account?	1	2	atoms4010002_perova	0
+Author Response The authors present a new interpretation to study Bloch oscillations in a cavity sustained optical lattice and the potential use of this system for very sensitive force measurement.The idea is closely related to previous work as Ref.			(1) It should be somehow mentioned more clearly (maybe in the abstract) that the idea is not completely new but work presents a new and more intutive picture of the system dynamics.	1	2	atoms4010002_perova	0
+This will generate extra noise, but might also help as it reduces atom number uncertainty in the cavity.			(2) It is very helpful to connect cavity output spectra and forces. To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40. This should be mentioned.	1	2	atoms4010002_perova	0
+(b) There will be heating limiting the available measurement time.			(3) The authors claim there should be no backaction of the field on the oscillation frequencies. Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil. This is somehow in contradiction with the eigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.	1	2	atoms4010002_perova	0
+This is somehow in contradiction with theeigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.			(b) There will be heating limiting the available measurement time. as the time scale can be faster than om_recoil, it could limit the measurement time to a fairly short period. Some rough estimate on the time scales should be added. In particular for the blue detuned case not only the average momentum but also the momentum spread will grow.	1	2	atoms4010002_perova	0
+* The fact that as the incident energy increases, the TDCS decreases and shifts towards smaller ejection angles for positron impact is said twice between lines 108 and 115.			The authors did not mention this method for discussions in the manuscript. In earlier publications [Physics Reports 315 (1999) 409 and related references], a fully relativistic distorted-wave Born approximation (rDWBA) has been developed to describe the relativistic ionization processes.	1	2	atoms4010010_makarova	0
+5 an 6 are mentioned but only 5 is discussed.			The RPWBA method in the manuscript should be compared with the previous rDWBA theory.	1	2	atoms4010010_makarova	0
+This research field, concerning the three-body dynamics, is known to be a well-studied area.			Again in earlier publications [Physics Reports 315 (1999) 409 and related references], there are existing experiments of triple differential cross sections for relativistic K-shell ionization of Ag, Au, and so on. The authors did not mention these experimental works in the manuscript.	1	2	atoms4010010_makarova	0
+"* Line 40: ""positrons"" -> ""positron"" * Line 43: ""positrons"" -> ""positron"" and ""ground state"" -> ""the ground state""."			(ii) There are repeated Bibliography like ref. 11 and ref. 31.	1	2	atoms4010010_makarova	0
+The work and discussion are very short, but still interesting.I recommend it for publication after the authors answer the questions and make the corrections described below:			In addition, I do believe that several figures could be combined in order to minimize journal space.	1	2	atoms4010010_makarova	0
+In spite of the lack of any experimental, or, as is claimed, any theoretical, verification of this or any physical interpretation as to why this should occur even at very high velocities, I believe that work deserves publication since it should incite additional theoretical attempts to verify these predictions.			"Page 9:* ""experimental one"" -> ""experimental ones"""	1	2	atoms4010010_makarova	0
+Thank you for taking the time and energy to help us improve the paper.			The authors did not mention this method for discussions in the manuscript. The RPWBA method in the manuscript should be compared with the previous rDWBA theory. At least the authors should mention the differences of the two methods and how far the RPWBA can be compared to rDWBA? In earlier publications [Physics Reports 315 (1999) 409 and related references], a fully relativistic distorted-wave Born approximation (rDWBA) has been developed to describe the relativistic ionization processes.	1	2	atoms4010010_perova	0
+"Maybe the authors could use ""is related to the probability"" or something like that."			Again in earlier publications [Physics Reports 315 (1999) 409 and related references], there are existing experiments of triple differential cross sections for relativistic K-shell ionization of Ag, Au, and so on. The authors did not mention these experimental works in the manuscript. It would be nice if the RPWBA method in the manuscript can be compared with the experiments.	1	2	atoms4010010_perova	0
+I find no major problems with the manuscript, either in content or presentation.			The authors have not carefully proofread this manuscript. Evidences of this includes (i) some errors in English typing, e.g. line 27 and line 119: it should be ‘plane wave …’ instead of ‘plan wave …’. Line between 33 and 34: it should be ‘The scattered positron and ejected electron …’ instead of ‘The scattered electron and ejected positron …’. (ii) There are repeated Bibliography like ref. 11 and ref. 31.	1	2	atoms4010010_perova	0
+"* Line 19: ""Coulomb system"" -> ""Coulomb systems"""			In addition, I do believe that several figures could be combined in order to minimize journal space.	1	2	atoms4010010_perova	0
+"------------------------------------------- Page 2: * Line 36: ""describes the probability"": actually it is the flux ratio, and it could be divergent in some cases."			"Page 9:* ""experimental one"" -> ""experimental ones"""	1	2	atoms4010010_perova	0
+Author response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			The first example is the result announced in Equation (4), concerning the phase shift due to the gravity gradient gamma. This effect was first calculated by Wolf and Tourrencin Physics Letters A 251 (1999) pp 241–246 and discussed with more details in A. Peters, K.Y. Chung and S. Chu, Metrologia 2001, 38, 25–61.	1	2	atoms4020014_perova	0
+I recommend accepting this manuscript, provided the following points are clarified:			Moreover, the authors call this effect a quantum correction probably because it is proportional to hbar but it would be more clear to explain that, the leading term of the gravitation phase being independent of hbar because of a cancellation (hbar appears in the numerator and denominator of the quantity giving the phase), the term linear in gamma is also proportional to the recoil velocity which itself is proportional to hbar.	1	2	atoms4020014_perova	0
+The results obtained seem reasonable, although I have no time to verify all the derivations.			A second example is the equation 9b for Wigner function: the associated quotation is ref. 17 by one of the authors and M.A. Kasevich in Phys Rev A (2006). A brief search on the web proves that a similar equation was used by Hongyi Fan Phys Rev A 656, 064102 (2002) who quotes two references dating from 1984 and a book published in 1994. I am not sufficiently aware of the use f Wigner function but I am sure that there are better and more useful references to quote in addition to ref 17!	1	2	atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			A second example is the equation 9b for Wigner function: the associated quotation is ref. 17 by one of the authors and M.A. Kasevich in Phys Rev A (2006). A brief search on the web proves that a similar equation was used by Hongyi Fan Phys Rev A 656, 064102 (2002) who quotes two references dating from 1984 and a book published in 1994. I am not sufficiently aware of the use f Wigner function but I am sure that there are better and more useful references to quote in addition to ref 17!	1	2	atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			In my opinion, phi_Q is the real quantum correction due to the spreading of the wavepacket during propagation. If I am correct, this should be explained.	1	2	atoms4020014_perova	0
+The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			The meaning of the three x_C, x_qC and x_QC (Equation (99)) is far from obvious but it is very surprising that the leading term in T^2 is different?	1	2	atoms4020014_perova	0
+The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			The paper contains only one figure with 28 panels presenting the results of the calculations. I think that this figure is very difficult to read! I do not understand clearly why results are plotted with y < y_min if this not physical: please explain!	1	2	atoms4020014_perova	0
+I recommend accepting this manuscript, provided the following points are clarified:			The paper contains only one figure with 28 panels presenting the results of the calculations. I think that this figure is very difficult to read! I do not understand clearly why results are plotted with y < y_min if this not physical: please explain!	1	2	atoms4020014_perova	0
+Authors’ response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			A figure explaining the geometry of the experiment would also help.	1	2	atoms4020014_perova	0
+1 Review Report 1 Originality/Novelty This paper treats an important question concerning the measurement of the gravitational constant G by atom interferometry.			The process is calculated as if the gravitational forces can be neglected during the diffraction laser pulses. This is probably an excellent approximation but please comment it!	1	2	atoms4020014_perova	0
+The results obtained seem reasonable, although I have no time to verify all the derivations.			The fountain is assumed to be exactly symmetric (see line after Equation (100)). It is well known that with symmetric fountains diffraction with retro-reflected laser beams produces stray interferometers... Minor Remarks	1	2	atoms4020014_perova	0
+Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			I noted an error after Equation (1): ... the gravitational potential is constant... I assume that the authors meant acceleration g, not the potential.	1	2	atoms4020014_perova	0
+I recommend accepting this manuscript, provided the following points are clarified:			The paper is quite difficult to read, because of very many bulky equations. I do not see how to reduce the number of equations but I think that some more comments would help the reader.	1	2	atoms4020014_perova	0
+Authors’ response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			The relative location of the test mass to the cold atom cloud should be clearly stated during the calculation, as the gravitational force is a vector. If the test mass is not located in the direction of the evolution trajectory of the atom cloud, the applicability of some formulas (e.g. Equation (3)) need to be verified. It will be helpful to the readers if a figure is provided to show the locations.	1	2	atoms4020014_perova	0
+These results are interesting to the people working in the field of atom interferometer, because the test mass are often used in the practical experiment.			The cloud is assumed to be characterized by a Wigner distribution function. What are the advantages to use Wigner distribution function instead of wave function?	1	2	atoms4020014_perova	0
+These results are interesting to the people working in the field of atom interferometer, because the test mass are often used in the practical experiment.			In the Introduction, quite a lot of papers are cited concerning the applications of atom interferometer in varies fields. Just for the note of the authors, there are some new applications of atom interferometer, e.g., in the accurate measurement of the quadratic Zeeman coefficient [J. Phys. B: At. Mol. Opt. Phys., 47(1): 015001, 2014].	1	2	atoms4020014_perova	0
+This EPS was carefully purified according to standard purification protocols for EPS.			The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.	1	2	biom12060834_makarova	0
+The studied EPS showed promising properties to use in the food industry.			Optimization of production constitutes an important part not mentioned in the title.	1	2	biom12060834_makarova	0
+Comment 15: l. 169-170: “thirty” instead of “thirteen”?			Acceptability for food applications was not discussed.	1	2	biom12060834_makarova	0
+Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.			Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.	1	2	biom12060834_makarova	0
+Response 2: Dear reviewer, thanks for your valuable comments for improving the quality of this paper.			EPS vs. EPSs in plural form should be consistent throughout.	1	2	biom12060834_makarova	0
+If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript.			Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.	1	2	biom12060834_makarova	0
+433, 434, 454, 458, and 507 : First section 3.6 (and its subsections) should be section 3.5.			l. 160 There is no coded value in Table 1.	1	2	biom12060834_makarova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			l. 165 Table 2 where Table 1 expected	1	2	biom12060834_makarova	0
+The following comments were not addressed correctly:			l. 169-170 thirty instead of thirteen ?	1	2	biom12060834_makarova	0
+Comment 22: Figure 1: What is the significance of the tick mark labels on the maps?			l. 177 i instead of 0 in second term?	1	2	biom12060834_makarova	0
+Comment 4: Acceptability for food applications was not discussed.			l. 228 GPC defined on l. 231	1	2	biom12060834_makarova	0
+Units are not specified on the axes labels or in the legend.			l. 230 PEG undefined	1	2	biom12060834_makarova	0
+Answer 25: Suggestion has been included in the revised version of the manuscript Comment 26: Figure 3: Units are not specified on the axes labels or in the legend.			l. 247 Volume of 0.2 mM ethanolic DPPH solution?	1	2	biom12060834_makarova	0
+The evidences for the partial structure given are not convincing.			255, 286, and 441 et al.	1	2	biom12060834_makarova	0
+Comment 2: The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.			Figure 1 What is the significance of the tick mark labels on the maps? Are these maps really necessary?	1	2	biom12060834_makarova	0
+Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			Figure 2 No x-axis label Units not specified	1	2	biom12060834_makarova	0
+Answer 1: To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript.			Figure 3	1	2	biom12060834_makarova	0
+Comment 22: Figure 1: What is the significance of the tick mark labels on the maps?			417 and 432 30.0 instead of 3.0 ?	1	2	biom12060834_makarova	0
+Comment 27: l. 414, Delete “(“before “C. pH”. ll. 416 and 432 , Add “C.” before “pH=” . ll. 417 and 432 , “30.0” instead of “3.0” ?			Units are not specified on the axes labels or in the legend. Figure 4 This figure is redundant as it gives the same information as Figure 3. Tick label values for glucose and yeast extract are different.	1	2	biom12060834_makarova	0
+Comment 34: The abbreviation for glucose is Glc.			433, 434, 454, 458, and 507 First section 3.6 (and its subsections) should be section 3.5.	1	2	biom12060834_makarova	0
+Fourier transform infrared spectroscopy (FTIR) and NMR spectroscopy data can only be regarded as preliminary.			l. 471, 600, 678, and 718 Reference format is different; corresponding references could be missing in the list.	1	2	biom12060834_makarova	0
+According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			l. 480 Why was linkage analysis not performed?	1	2	biom12060834_makarova	0
+This allows us to conclude they are different polysaccharides.			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	0
+(I was referring to the chemical method by methylation.)			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	0
+Comment 42: l. 683-684: Food-grade oils are not aliphatic and aromatic hydrocarbons.			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	0
+According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_makarova	0
+Comment 35: l. 500 : The chemical shift range is more than from 1 to 5.			l. 500 The chemical shift range is more than from 1 to 5. ll.	1	2	biom12060834_makarova	0
+EPC is isolated from a thermal spring of volcanic origin, which contains a set of various elements, including heavy metals.			l. 537 No section 3.6.2	1	2	biom12060834_makarova	0
+Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			l. 636 Glc and Gal	1	2	biom12060834_makarova	0
+We will answer in the following paragraphs because all comments were related to the same topic.			l. 683-684 Food-grade oils are not aliphatic and aromatic hydrocarbons.	1	2	biom12060834_makarova	0
+EPC is isolated from a thermal spring of volcanic origin, which contains a set of various elements, including heavy metals.			l. 725 possibility of future	1	2	biom12060834_makarova	0
+Comment 1: Unfortunately, its structure was not fully determined.			The English language was improved, but there are still errors.	3	2	biom12060834_makarova	0
+Comment 42: l. 683-684: Food-grade oils are not aliphatic and aromatic hydrocarbons.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+Answer 29: As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript Comment 30: l. 442: “C), whereas the” Answer 30: Suggestion has been included in the revised version of the manuscript Comment 31: l. 471, 600, 678, and 718: Reference format is different; corresponding references could be missing in the list.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+Bacterial EPSs are normally composed of repeating units.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+These results suggest they are the prevailing units of the EPS backbone making it a branched one.			Why was linkage analysis not performed? Bacterial EPSs are normally composed of repeating units. Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned. (I was referring to peak intensities that are not uniform. The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.) The authors added a COSY spectrum but did not analyze it. Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. (I was referring to the chemical method by methylation.). They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_makarova	0
+We will answer in the following paragraphs because all comments were related to the same topic.			To my deep regret the structural characteristic of EPS is the weak point of this publication.	1	2	biom12060834_makarova	0
+528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.	1	2	biom12060834_makarova	0
+Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			"Line 42: It is premature to include the term ""structure"" in keywords at this stage."	1	2	biom12060834_makarova	0
+See file Comments for author File: Comments.pdf			Line 526-527: The proposal should be reformulated. According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.	1	2	biom12060834_makarova	0
+Comment 22: Figure 1: What is the significance of the tick mark labels on the maps?			The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.	1	2	biom12060834_perova	0
+481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			Optimization of production constitutes an important part not mentioned in the title.	1	2	biom12060834_perova	0
+Tick label values for glucose and yeast extract are different.			Acceptability for food applications was not discussed.	1	2	biom12060834_perova	0
+Comment 27: l. 414, Delete “(“before “C. pH”. ll. 416 and 432 , Add “C.” before “pH=” . ll. 417 and 432 , “30.0” instead of “3.0” ?			English language should be revised thoroughly. There are inconsistencies in singular-plural concordance between subject and verb as well as noun and pronoun. Verb tenses should be checked. Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.	1	2	biom12060834_perova	0
+50-51: “improvement of rheological” Answer 8:			EPS vs. EPSs in plural form should be consistent throughout.	1	2	biom12060834_perova	0
+TOCSY and HMBC are mentioned in methods, but no results are presented.			Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.	1	2	biom12060834_perova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			l. 160 There is no coded value in Table 1.	1	2	biom12060834_perova	0
+How can the authors ascertain that mannose does not come from mannans in yeast extract?			l. 165 Table 2 where Table 1 expected	1	2	biom12060834_perova	0
+Units are not specified on the axes labels or in the legend.			l. 169-170 thirty instead of thirteen ?	1	2	biom12060834_perova	0
+The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			l. 177 i instead of 0 in second term?	1	2	biom12060834_perova	0
+The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			l. 228 GPC defined on l. 231	1	2	biom12060834_perova	0
+Figure 6: Bacterial EPSs are normally composed of repeating units.			l. 230 PEG undefined	1	2	biom12060834_perova	0
+528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			l. 247 Volume of 0.2 mM ethanolic DPPH solution?	1	2	biom12060834_perova	0
+Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit.			255, 286, and 441 et al.	1	2	biom12060834_perova	0
+The studied EPS showed promising properties to use in the food industry.			Figure 1 What is the significance of the tick mark labels on the maps? Are these maps really necessary?	1	2	biom12060834_perova	0
+Comment 4: Acceptability for food applications was not discussed.			Figure 2 No x-axis label Units not specified	1	2	biom12060834_perova	0
+Comment 7: Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.			Figure 3	1	2	biom12060834_perova	0
+TOCSY and HMBC are mentioned in methods, but no results are presented.			417 and 432 30.0 instead of 3.0 ?	1	2	biom12060834_perova	0
+The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			Units are not specified on the axes labels or in the legend. Figure 4 This figure is redundant as it gives the same information as Figure 3. Tick label values for glucose and yeast extract are different.	1	2	biom12060834_perova	0
+528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			433, 434, 454, 458, and 507 First section 3.6 (and its subsections) should be section 3.5.	1	2	biom12060834_perova	0
+The authors added a COSY spectrum but did not analyze it.			l. 471, 600, 678, and 718 Reference format is different; corresponding references could be missing in the list.	1	2	biom12060834_perova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			l. 480 Why was linkage analysis not performed?	1	2	biom12060834_perova	0
+Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	0
+Comment 21: l. 284: No section 2.6.3 Answer 21:			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	0
+The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.)			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	0
+→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			Figure 6 Bacterial EPSs are normally composed of repeating units. How can the authors ascertain that mannose does not come from mannans in yeast extract? A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll. 495-496 is wrong). The abbreviation for glucose is Glc.	1	2	biom12060834_perova	0
+Comment 14: l. 165: Table 2 where Table 1 expected Answer 14:			l. 500 The chemical shift range is more than from 1 to 5. ll.	1	2	biom12060834_perova	0
+Comment 16: l. 177: “βi” instead of “β0” in second term?			l. 537 No section 3.6.2	1	2	biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			l. 636 Glc and Gal	1	2	biom12060834_perova	0
+→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			l. 683-684 Food-grade oils are not aliphatic and aromatic hydrocarbons.	1	2	biom12060834_perova	0
+Comment 16: l. 177: “βi” instead of “β0” in second term?			l. 725 possibility of future	1	2	biom12060834_perova	0
+Comment 39: l. 581: “Table S1A” Answer 39: Suggestion has been included in the revised version of the manuscript Comment 40: l. 603: “Figure S1” Answer 40: Suggestion has been included in the revised version of the manuscript Comment 41: l. 636: “Glc and Gal” Answer 41:			The English language was improved, but there are still errors.	3	2	biom12060834_perova	0
+Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Author Response Please see the attachment for the responses.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+→ Comment 1: The English language was improved, but there are still errors.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Comment 34: The abbreviation for glucose is Glc.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Comment 15: l. 169-170: “thirty” instead of “thirteen”?			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Author Response Please see the attachment to see the responses.			Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit. The evidences for the partial structure given are not convincing. If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript. They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.The information contained in the other 2D spectra was not exploited to provide structural information.	3	2	biom12060834_perova	0
+Comment 4: Acceptability for food applications was not discussed.			To my deep regret the structural characteristic of EPS is the weak point of this publication. Fourier transform infrared spectroscopy (FTIR) and NMR spectroscopy data can only be regarded as preliminary. According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure. Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc. ), as well as two-dimensional NMR spectroscopy.	1	2	biom12060834_perova	0
+Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			EPC is isolated from a thermal spring of volcanic origin, which contains a set of various elements, including heavy metals. It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.	1	2	biom12060834_perova	0
+Comment 34: The abbreviation for glucose is Glc.			"Line 42: It is premature to include the term ""structure"" in keywords at this stage."	1	2	biom12060834_perova	0
+Basically, the work was done at a fairly high professional level, the material is presented clearly, well structured, the data obtained are discussed with the previously obtained literature data.			Line 526-527: The proposal should be reformulated. According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.	1	2	biom12060834_perova	0
+We would prefer to include those results in the upcoming publication.			It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	0
+The same cDNA amount for every condition was used to perform qRT-PCR assays.			It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	0
+https://www.sciencedirect.com/science/article/abs/pii/S0960894X08011086?via%3Dihub			It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	0
+"The authors presented the paper ""Human Serum Proteins and Susceptibility of Acinetobacter baumannii to Cefiderocol: role of iron transport""."			It is known that HSA can bind RNA and DNA and has RNA hydrolyzing activity. In this way your experiment in par 2.2. may lead to not all RNA after incubation with such amount of albumin. That is why some of your effects may have other explanations.	1	2	biomedicines10030600_makarova	0
+If we understand correctly, the reviewer’s argument, i.e., a reduction in RNA when the cells were cultured in the presence of HSA may be due to the HSA’s RNA-hydrolyzing activity, requires that HSA contacts the RNA that will be hydrolyzed.			Cefiderocol is a cidal antibiotic. It would be of great interest if for the key results the authors would also provide MBC and / or time-resolved kill-curves. I believe this kind of data would be of great relevance for assessing the clinical relevance of the observed effects.	1	2	biomedicines10030600_makarova	0
+Patient symptoms likely arise from mutations disturbing the role that the encoded NMDA receptor subunit, GluN2B, plays at neuronal connections in the developing nervous system.			Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative. It follows from the methodology that AMPA currents were recorded on a regular basis.	1	2	brainsci12060789_makarova	0
+1) Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative.			Please add a chart about tau changes in Figure 3.	1	2	brainsci12060789_makarova	0
+In this study, we have investigated the cell-autonomous effects of putative gain- (GoF) and loss-of-function (LoF) missense GRIN2B mutations on excitatory synapses onto CA1 pyramidal neurons in organotypic hippocampal slices.			Please verify if the time scale is correct in Fig. 3А.	1	2	brainsci12060789_makarova	0
+Our results demonstrate that functional incorporation of GoF and LoF GluN2B mutants into synaptic receptors and the effects on EPSC decay times are highly dependent on the presence of triheteromeric GluN1/2A/2B NMDA receptors, thereby influencing the functional classification of NMDA receptor variants as GoF or LoF mutations.			A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion	1	2	brainsci12060789_makarova	0
+Please verify if the time scale is correct in Fig. 3А.			A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.	1	2	brainsci12060789_makarova	0
+Author Response Thank you to the reviewer for their comments and helpful suggestions.			Even though fig.2aiii shows a comparable relative peak amplitude, which indicates both GOF and LOF rescue NMDA-EPSC in Grin2b-/- neurons, it looks to me that the absolute amplitude in fig.2aii shows a quite big difference in un-transferred groups which are supposed to be comparable, not as consistent as shown in fig.1ci. How to explain this discrepancy?	1	2	brainsci12060789_makarova	0
+The authors conclude that the presence of triheteromeric NMDA receptors, and the types of the mutated GluN2 subunit are all important factors that can influence the function of the NMDA variants.			The supplementary figures are inaccessible.	1	2	brainsci12060789_makarova	0
+There are also a number of sentences that are so complicated for easy read.			The abstract is too long, which does not actually abstract the whole content very well. There are also a number of sentences that are so complicated for easy read. This part should be substantially polished into a shortened and precise style.	1	2	brainsci12060789_makarova	0
+Please find below our point-by-point response.			Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative. It follows from the methodology that AMPA currents were recorded on a regular basis.	1	2	brainsci12060789_perova	0
+The abstract is too long, which does not actually abstract the whole content very well.			Please add a chart about tau changes in Figure 3. Please verify if the time scale is correct in Fig. 3А.	1	2	brainsci12060789_perova	0
+Please find below our point-by-point response.			A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion	1	2	brainsci12060789_perova	0
+Author Response Thank you to the reviewer for their comments and helpful suggestions.			A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.	1	2	brainsci12060789_perova	0
+1) Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative.			Even though fig.2aiii shows a comparable relative peak amplitude, which indicates both GOF and LOF rescue NMDA-EPSC in Grin2b-/- neurons, it looks to me that the absolute amplitude in fig.2aii shows a quite big difference in un-transferred groups which are supposed to be comparable, not as consistent as shown in fig.1ci. How to explain this discrepancy?	1	2	brainsci12060789_perova	0
+Please find below our point-by-point response.			The supplementary figures are inaccessible.	1	2	brainsci12060789_perova	0
+There are some concerns need to be addressed.			The abstract is too long, which does not actually abstract the whole content very well. There are also a number of sentences that are so complicated for easy read. This part should be substantially polished into a shortened and precise style.	1	2	brainsci12060789_perova	0
+This manuscript describes a cohort randomized crossover design comparing cutaneous heat pain ratings and brain activity during heat stimuli after conditions of quiet rest and cycling in patients with a physician-confirmed diagnosis of Fibromyalgia (FM) and age and sex matched controls.			However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls.	1	2	brainsci6010008_makarova	0
+The time post-exercise before scanning was provided, but not the time post-quiet rest.			The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_makarova	0
+This study has compared acute effects of exercise and inactivity in fibromyalgia (FM) and pain-free controls on changes in pain and cerebral activity in response to heat.			Also, the discussion section should be revised to acknowledge that many patients with FM would have been excluded from participation in this study due to major depression and medication consumption so there may be concerns about the generalizability of the sample, BUT the authors appropriately compared the Fibromyalgia Impact Questionnaire scores of their sample to normative data and found no significant difference.	1	2	brainsci6010008_makarova	0
+“ Each temperature was applied twice so I assume averages were used, but the authors should clarify.			In addition, the authors might want to consider and mention that their method of controlling “physiological noise” might have influenced the results because cardiovascular reactions to exercise were, at one time, proposed to be a mechanism of EIH.	1	2	brainsci6010008_makarova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			P. 1, L. 40 – I don’t see that the reference actually includes studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	1	2	brainsci6010008_makarova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			P. 2, L. 18 – A reference for the determination that a dosage of antidepressants was “high” should be provided.	1	2	brainsci6010008_makarova	0
+Considering discussion the results in relation to previous studies on exercise and neuroimaging.			P. 3, L. 15 – The time post-exercise before scanning was provided, but not the time post-quiet rest.	1	2	brainsci6010008_makarova	0
+The results are clearly reported and adequately discussed.			P. 6, L. 5 – “Elevations” should be revised to “higher” so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.	1	2	brainsci6010008_makarova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			P. 6, L. 12 - I am uncertain why the authors only report the effect sizes of group differences for the first run.s	1	2	brainsci6010008_makarova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_makarova	0
+Nine individuals in each group were indicated in the neuroimaging analysis, this should be indicated in the abstract.			Nine individuals in each group were included in neuroimaging analyses, this should be indicated in the abstract.	1	2	brainsci6010008_makarova	0
+Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			"In table 3 the subheading ""Peak X, Y, X"" needs correction."	1	2	brainsci6010008_makarova	0
+Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			Consider discussing the results in relation to previous studies on exercise and neuroimaging	1	2	brainsci6010008_makarova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			The findings are novel and may be compared to previous studies of exercise and neuroimaging in fibromyalgia.	1	2	brainsci6010008_makarova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls. The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_perova	0
+Nine individuals in each group were indicated in the neuroimaging analysis, this should be indicated in the abstract.			However, the authors need to compare their findings to other research evidence showing impaired EIH and/or descending inhibition in patients with FM - especially considering that their results did not support EIH in the controls. The authors merely mention that the exercise stimulus may have been insufficient for the controls to show EIH without critically discussing what such a conclusion would mean (eg, FM patients have better EIH mechanisms than the controls).	1	2	brainsci6010008_perova	0
+The methodology is sound and the manuscript is well-written.			Also, the discussion section should be revised to acknowledge that many patients with FM would have been excluded from participation in this study due to major depression and medication consumption so there may be concerns about the generalizability of the sample, BUT the authors appropriately compared the Fibromyalgia Impact Questionnaire scores of their sample to normative data and found no significant difference.	1	2	brainsci6010008_perova	0
+We feel that the manuscript has been significantly improved as a result of the suggested revisions.			In addition, the authors might want to consider and mention that their method of controlling “physiological noise” might have influenced the results because cardiovascular reactions to exercise were, at one time, proposed to be a mechanism of EIH.	1	2	brainsci6010008_perova	0
+In table 3, the subheading “Peak X, Y, X” needs correction.			P. 1, L. 40 – I don’t see that the reference actually includes studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	1	2	brainsci6010008_perova	0
+The methodology is sound and the manuscript is well-written.			P. 2, L. 18 – A reference for the determination that a dosage of antidepressants was “high” should be provided.	1	2	brainsci6010008_perova	0
+The time post-exercise before scanning was provided, but not the time post-quiet rest.			P. 3, L. 15 – The time post-exercise before scanning was provided, but not the time post-quiet rest.	1	2	brainsci6010008_perova	0
+The discussion interprets the results as supporting evidence of exercise-induced hypoalgesia (EIH) in the patients with FM.			P. 6, L. 5 – “Elevations” should be revised to “higher” so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.	1	2	brainsci6010008_perova	0
+A reference for the determination that a dosage of antidepressants was “high” should be provided.			P. 6, L. 12 - I am uncertain why the authors only report the effect sizes of group differences for the first run.s	1	2	brainsci6010008_perova	0
+This manuscript describes a cohort randomized crossover design comparing cutaneous heat pain ratings and brain activity during heat stimuli after conditions of quiet rest and cycling in patients with a physician-confirmed diagnosis of Fibromyalgia (FM) and age and sex matched controls.			However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_perova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			However, in the latter case the authors need to address the issue of multiple correlations. Changes in pain sensitivity after exercise versus rest were significantly correlated with changes in activity in DLPFC (exercise vs. rest).	1	2	brainsci6010008_perova	0
+We feel that the manuscript has been significantly improved as a result of the suggested revisions.			Nine individuals in each group were included in neuroimaging analyses, this should be indicated in the abstract.	1	2	brainsci6010008_perova	0
+This study has compared acute effects of exercise and inactivity in fibromyalgia (FM) and pain-free controls on changes in pain and cerebral activity in response to heat.			"In table 3 the subheading ""Peak X, Y, X"" needs correction."	1	2	brainsci6010008_perova	0
+Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			Consider discussing the results in relation to previous studies on exercise and neuroimaging	1	2	brainsci6010008_perova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			The results are clearly reported and adequately discussed. The findings are novel and may be compared to previous studies of exercise and neuroimaging in fibromyalgia.	1	2	brainsci6010008_perova	0
+Are there better references to substantiate this point?			"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."	1	2	brainsci6010009_makarova	0
+On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			In the exposed and control groups were there other related diseases such as Autism? If so, how many and what would happen to the analysis if you removed them?	1	2	brainsci6010009_makarova	0
+The writing is clear, the analysis is sound, and the presentation is excellent with a few exceptions.			On page 3 line 22, they describe children with the diagnosis before the exposure. How many were there that fit in this category?	1	2	brainsci6010009_makarova	0
+How many were there that fit in this category?			On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point. Are there better references to substantiate this point?	1	2	brainsci6010009_makarova	0
+Can the authors address this point: is this database publicly available?			Can the authors address this point: is this database publicly available?	1	2	brainsci6010009_makarova	0
+Are there better references to substantiate this point?			The results are controversial. However, the findings may be of great interest for the readers of the journal. The paper is of high quality. I recommend the manuscript published. It can be published as it is.	1	2	brainsci6010009_makarova	0
+On page 3 line 22, they describe children with the diagnosis before the exposure.			"1. In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."	1	2	brainsci6010009_perova	0
+The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			2. In the exposed and control groups were there other related diseases such as Autism? If so, how many and what would happen to the analysis if you removed them?	1	2	brainsci6010009_perova	0
+These results are relevant and important as thimerosal is still included in global vaccines although they have largely been removed from childhood vaccine in the United States.			3. On page 3 line 22, they describe children with the diagnosis before the exposure. How many were there that fit in this category?	1	2	brainsci6010009_perova	0
+The writing is clear, the analysis is sound, and the presentation is excellent with a few exceptions.			On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point. Are there better references to substantiate this point?	1	2	brainsci6010009_perova	0
+If so, how many and what would happen to the analysis if you removed them?			5. Can the authors address this point: is this database publicly available?	1	2	brainsci6010009_perova	0
+"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."			This is an important study that deserves to be published. The results are controversial. However, the findings may be of great interest for the readers of the journal. The paper is of high quality. I recommend the manuscript published. It can be published as it is.	1	2	brainsci6010009_perova	0
+Round 1: Author Response to Reviewer 2 In my view, this manuscript will make a nice contribution to the literature.			This is a well-written paper with a large sample of girls that replicates and extends important research on relational aggression. The focus on the functions (reactive and proactive) of relational aggression has significant implications for intervention with adolescent girls. I commend the authors for this timely and crucial study on factors that influence the development of aggression and antisocial behavior in girls, which is typically an understudied topic.	1	2	bs5040518_makarova	0
+Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.			First, it is recommended that the authors provide more information about the selection of the cluster solution. That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth. It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.	1	2	bs5040518_makarova	0
+That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth.			Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.	1	2	bs5040518_makarova	0
+Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model.			First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue.	1	2	bs5040518_makarova	0
+These findings are in contrast to earlier work, which assumed that proactive and reactive aggression represented distinct typologies.			Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_makarova	0
+Below are a few comments that could hopefully help improve a solid piece of work.			Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.	1	2	bs5040518_makarova	0
+Round 1: and Author Response The goal of this study was to distinguish subgroups of relationally aggressive females (i.e., reactively aggressive-only subgroup, reactively-and-proactively aggressive-combined subgroup, non-aggressive subgroup), on the basis of nine factors reflecting individual characteristics, peer-related variables and R3 parental control.			Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.	1	2	bs5040518_makarova	0
+Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.			A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).	1	2	bs5040518_makarova	0
+Acknowledging this problem in the limitations does not solve it.			Some findings are difficult to reconcile with the current literature; in addition, they are internally inconsistent. For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?	1	2	bs5040518_makarova	0
+Acknowledging this problem in the limitations does not solve it.			There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.	1	2	bs5040518_makarova	0
+Overall, this is a nice paper that looks to make a contribution.			There are enough variables that I hard a hard time following the analyses. A tighter focus in the intro and matching of analyses to hypotheses may help here.	1	2	bs5040518_makarova	0
+This work, as with other recent work, makes clear that the most severely aggressive youth tend to engage in high R2 levels of both proactive and reactive aggression and are otherwise also generally more distressed and dysregulated, as compared with youth who are more moderately aggressive and tend to engage in reactive aggression only, and youth who have low levels of aggression.			This is a well-written paper with a large sample of girls that replicates and extends important research on relational aggression. The focus on the functions (reactive and proactive) of relational aggression has significant implications for intervention with adolescent girls. I commend the authors for this timely and crucial study on factors that influence the development of aggression and antisocial behavior in girls, which is typically an understudied topic.	1	2	bs5040518_perova	0
+Below are a few comments that could hopefully help improve a solid piece of work.			First, it is recommended that the authors provide more information about the selection of the cluster solution. That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth. It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.	1	2	bs5040518_perova	0
+Despite the use of a relatively large sample and of sound measures, this study falls short of making an important contribution to the current literature for several reasons.			Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.	1	2	bs5040518_perova	0
+There are two areas to address to strengthen the paper a bit more.			Although the rationale for selecting each of the 9 factors that might be differentially related to reactive or proactive relational aggression is convincing, the overall picture is incomplete on at least two accounts. First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue. Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_perova	0
+There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.			Although the rationale for selecting each of the 9 factors that might be differentially related to reactive or proactive relational aggression is convincing, the overall picture is incomplete on at least two accounts. First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression. If the authors are really interested in the processes that could feed into reactive or proactive relational aggression in females, they should have been more attentive to this issue. Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model (i.e., peer and parent factors potentiate the link between individual factors and subtypes of relational aggression).	1	2	bs5040518_perova	0
+The study is strengthened by the person-centered approach to data analysis, by the large sample, by focusing on girls who have been much less studied than boys, and by the clear rationale for the study.			The use of a cross-sectional design is a major limitation, because it cannot help determine the directionality of the links between the nine factors and subtypes of relational aggression, left alone causality. This bears directly on the issue of predictors vs. correlates vs. consequences of subtypes of relational aggression. Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.	1	2	bs5040518_perova	0
+This work, as with other recent work, makes clear that the most severely aggressive youth tend to engage in high R2 levels of both proactive and reactive aggression and are otherwise also generally more distressed and dysregulated, as compared with youth who are more moderately aggressive and tend to engage in reactive aggression only, and youth who have low levels of aggression.			All the measures are self-reported. This artificially inflates the link between the study variables. Acknowledging this problem in the limitations does not solve it. Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.	1	2	bs5040518_perova	0
+For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			Creating groups based on a cluster analysis may have created unnecessary problems. In particular, and contrary to what the authors seem to believe, it is not possible to know whether differences between the two aggressive groups reflect differences in levels of relational aggression or in type of relational aggression (reactive only vs. combined), given that the combined group is obviously much more aggressive than the reactive-only group. A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).	1	2	bs5040518_perova	0
+Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.			Some findings are difficult to reconcile with the current literature; in addition, they are internally inconsistent. For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?	1	2	bs5040518_perova	0
+That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth.			There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.	1	2	bs5040518_perova	0
+Round 1: and Author Response This manuscript reports on person-centered analysis of adolescent girls finding that they are distinguished by types of aggression, callous-unemotional traits, and by interpersonal relationships with peers and parents.			There are enough variables that I hard a hard time following the analyses. A tighter focus in the intro and matching of analyses to hypotheses may help here.	1	2	bs5040518_perova	0
+It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.			As it is, this reviewer had difficulty following the variables as predictors/outcomes and the hypotheses that are driving the paper 2.	1	2	bs5040518_perova	0
+That seems to be the model implied by the analyses, but it is never explicitly specified.			With better theoretical framing and a more limited set of analyses, the value of the study results should be more clear.	1	2	bs6010004_makarova	0
+"**Thank you catching the typographical error (80% attracted to the same sex) - this should have read ""other sex""."			Also, a more thorough description of the measurement of the construct and the validity of the measurement could be described in that section.	1	2	bs6010004_makarova	0
+Fifth, one set of results seems quite contradictory: sense of entitlement to sexual self-pleasure decreased, on average, from Time 1 to Time 2, but sense of entitlement to sexual self-pleasure was positively correlated with age.			On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” **	1	2	bs6010004_makarova	0
+Participants completed two waves of data collection approximately 1-year apart.			The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.	1	2	bs6010004_makarova	0
+On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.			The authors rely too much on headings to transition between ideas. The paper would flow more smoothly if some transition sentences were included.	1	2	bs6010004_makarova	0
+What were participants told about the purpose of the study?			More specific hypotheses could be proposed.	1	2	bs6010004_makarova	0
+List-wise deletion can lead to serious bias in results.			The section where research questions are described is worded in an awkward way.	1	2	bs6010004_makarova	0
+There is an increase in Type I error with the multiple t-tests conducted thatshould be addressed.			The reporting of group differences in the participants section might be better placed in the results section after the measures are described.	1	2	bs6010004_makarova	0
+Third, although longitudinal data can be a quite powerful tool for understanding change and development, the current analyses squander some of that power.			The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained. Perhaps factor analysis could be used to determine appropriate subscales?	1	2	bs6010004_makarova	0
+From the relatively high percentage of non-heterosexual participants, there seems to be some selection bias.			What were participants told about the purpose of the study?	1	2	bs6010004_makarova	0
+"Overall, 91% were white/Caucasian, 3% were Asian, 1 was Aboriginal/Pacific Islander, and the remaining participants indicated an ""Other"" sociocultural background."			Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.	1	2	bs6010004_makarova	0
+The first paragraph of the Participants section now reads: The participants were 295 adolescent and young adult men (n = 112) and women (n = 183) aged between 17 to 25 years (M = 19.5 years, SD = 1.9).			Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).	1	2	bs6010004_makarova	0
+"Second, potential participants were ""approached at a university campus"" during orientation week."			Analyses should be re-done using a more sophisticated method for handling missing data, such as multiple imputation or full-information maximum likelihood (Shafer & Graham, 2002, Psychological Methods).	1	2	bs6010004_makarova	0
+Fifth, one set of results seems quite contradictory: sense of entitlement to sexual self-pleasure decreased, on average, from Time 1 to Time 2, but sense of entitlement to sexual self-pleasure was positively correlated with age.			At the very least, the Discussion should consider how generalizable these results are.	1	2	bs6010004_makarova	0
+The paper would flow more smoothly if some transition sentences were included.			More generally, I think the putative timescale of effects of behavior on sexual subjectivity needs more theoretical elaboration. Are behaviors in early adolescence (before age 16), for example, expected to continue to influence the trajectory of sexual subjectivity regardless of later sexual behavior?	1	2	bs6010004_makarova	0
+There was no evidence that any association differed between young men and young women.			The shift to a latent factor approach would have the added benefit of allowing the authors to use FIML to account for missing data at T2, and they could thus use their entire sample.	1	2	bs6010004_makarova	0
+"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and"			This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.	1	2	bs6010004_makarova	0
+**We also expanded the first paragraph of the Limitations to say: Although this study provided insight into how aspects of sexual subjectivity differed over one year in young men and women and uncovered associations of age and sexual behavior with sexual subjectivity, there were two limitations worthy of note.			I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion.	1	2	bs6010004_makarova	0
+Moreover, the reverse paths -- from sexual subjectivity to future behavior -- are also not tested in this paper.			my main concern with the study is that there were too many analyses presented to interpret in a meaningful way. The authors should revise the paper providing more theoretical background and accompanying analyses. With better theoretical framing and a more limited set of analyses, the value of the study results should be more clear.	1	2	bs6010004_perova	0
+First, the attrition between Time 1 and Time 2 was substantial (40% of participants), and this is handled using list-wise deletion.			I think it would be preferable to describe the measurement of sexual subjectivity shortly after it is defined on p. 3. Also, a more thorough description of the measurement of the construct and the validity of the measurement could be described in that section.	1	2	bs6010004_perova	0
+A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” **	1	2	bs6010004_perova	0
+Author Response Please specify how many students were approached to participate in the study and therefore specify the response rate.			The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.	1	2	bs6010004_perova	0
+There was no evidence that any association differed between young men and young women.			The authors rely too much on headings to transition between ideas. The paper would flow more smoothly if some transition sentences were included.	1	2	bs6010004_perova	0
+The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained.			More specific hypotheses could be proposed. The section where research questions are described is worded in an awkward way.	1	2	bs6010004_perova	0
+Therefore, the results of the current study may have some limited generalizability.			The reporting of group differences in the participants section might be better placed in the results section after the measures are described.	1	2	bs6010004_perova	0
+Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).			The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained. Perhaps factor analysis could be used to determine appropriate subscales?	1	2	bs6010004_perova	0
+The normative development of sexual health is an important and understudied topic, and the Introduction to this paper provides a good overview of the theoretical work in this area.			What were participants told about the purpose of the study?	1	2	bs6010004_perova	0
+There might be some interesting data collected in the present study.			There is an increase in Type I error with the multiple t-tests conducted thatshould be addressed. Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.	1	2	bs6010004_perova	0
+Some more specific suggestions are provided below.			Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).	1	2	bs6010004_perova	0
+How do the authors make sense of these result, as participants are one year older at Time 2?			First, the attrition between Time 1 and Time 2 was substantial (40% of participants), and this is handled using list-wise deletion. That is, any one who did not complete both assessments was not included in the analyses. List-wise deletion can lead to serious bias in results. Analyses should be re-done using a more sophisticated method for handling missing data, such as multiple imputation or full-information maximum likelihood (Shafer & Graham, 2002, Psychological Methods).	1	2	bs6010004_perova	0
+**We did not keep an exact count of the number of students approached about the study, but we estimate that we approached about 375 students.			"Second, potential participants were ""approached at a university campus"" during orientation week. What percentage of people who were approached agreed to participate? How broadly representative of the university population (or the university-age population) is the sample? From the relatively high percentage of non-heterosexual participants, there seems to be some selection bias. At the very least, the Discussion should consider how generalizable these results are."	1	2	bs6010004_perova	0
+Statistics show that the majority of Australian year 10-12 students (approximately 15-17 years) have engaged in some form of sexual behavior [12].			Third, although longitudinal data can be a quite powerful tool for understanding change and development, the current analyses squander some of that power. The regression results presented in Table 2 test whether sexual behaviors that participants already experienced by Time 1 predicted facets of sexual subjectivity at Time 2, controlling for sexual subjectivity at Time 1. Why would behavior that has already happened contribute to a re-ordering of individuals over the course of the next year? It seems that a more interesting and direct test of the longitudinal effects of sexual behavior on sexual subjectivity would test whether new sexual behaviors (that is, sexual behaviors experienced between Time 1 and Time 2) predict change in sexual subjectivity from Time 1 to Time 2. Moreover, the reverse paths -- from sexual subjectivity to future behavior -- are also not tested in this paper. More generally, I think the putative timescale of effects of behavior on sexual subjectivity needs more theoretical elaboration. Are behaviors in early adolescence (before age 16), for example, expected to continue to influence the trajectory of sexual subjectivity regardless of later sexual behavior? That seems to be the model implied by the analyses, but it is never explicitly specified.	1	2	bs6010004_perova	0
+Most lived with their parents (61%) and reported being only attracted to the other sex (80%).			Fourth, each analysis is conducted for each subscale separately, but (with the exception of sexual body-esteem) the scales are consistently (if moderately) intercorrelated. I think it would be informative to test whether associations with age or sexual experience group are operating through a general underlying factor of sexual subjectivity versus are unique to specific facets. The shift to a latent factor approach would have the added benefit of allowing the authors to use FIML to account for missing data at T2, and they could thus use their entire sample.	1	2	bs6010004_perova	0
+"**Thank you catching the typographical error (80% attracted to the same sex) - this should have read ""other sex""."			Fifth, one set of results seems quite contradictory: sense of entitlement to sexual self-pleasure decreased, on average, from Time 1 to Time 2, but sense of entitlement to sexual self-pleasure was positively correlated with age. How do the authors make sense of these result, as participants are one year older at Time 2? This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.	1	2	bs6010004_perova	0
+I have four recommendations for improving the paper.			"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion."	1	2	bs6010004_perova	0
+Author Response Response to Reviewer 1 Comments I would like to thank the authors for taking all my comments into full consideration.			The definition of biophilia is described as an “inherent love” toward nature. While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.	1	2	buildings12040417_makarova	0
+Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32.			P1, L39-41: Please provide references to these frameworks. Reading further to page 3, I believe these are the 24 biophilic design attributes [ref. 25,39], and the 14 patterns of biophilic design [ref.40]. Further references around the biophilic concept could also be provided, e.g.: Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. Ulrich, 1993. Biophilia, biophobia, and natural landscapes. Ko et al., 2020. A window view quality assessment framework. LEUKOS. The latter reference reviewed many international standards that advocate nature and biophilic design for view and building spaces, with examples given to the Singapore context. This somewhat overlaps with my next comment.	1	2	buildings12040417_makarova	0
+"3, Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."			P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment. WELL v2 has several features for Nature and Mind, and Biophilia – Parts I and II, with quantitative assessment methods provided. Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief. If the authors agree with this, perhaps this could be revised here to reflect this. Other standards likely incorporate biophilic elements in building architecture, and could be worth highlighting. The general issue raised by the authors do not necessarily imply a lack of guidelines for biophilic design, since there are several readily available, but may point toward prioritisation or emphasis of criteria to meet certain varying expectations, which was alluded to on lines 52-53.	1	2	buildings12040417_makarova	0
+Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief.			P2, L63: Although I wouldn’t completely rule this out, POE surveys may not always provide feedback to the architect, since they are implemented post design-stage and the building would be operated by facility management or the owner. In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues. Lessons learned from 20 years of CBE’s occupant surveys. Building & Cities. Kent et al., 2021. A data-driven analysis of occupant workspace dissatisfaction. Building and Environment. Cheung et al. 2021. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. Recently POE studies, also using office data, advocate this as benefit to their implementation, albeit not necessarily being the only reason: Graham et al., 2020.	1	2	buildings12040417_makarova	0
+This becomes an issue later, since some aspects referring to biophilic design become unclear.			P3, L92: I think refers to “has helped” given the five decades predating this.	1	2	buildings12040417_makarova	0
+Response 7: The method of selection of the biophilic design attributes/patterns for workplace is: Step one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks.			P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation. Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32. Therefore, it would be more appropriate to use psychological scales (e.g., PANAS or psychological restoration), instead of design orientated question or survey. If the authors agree with this, this aspect could be revised.	1	2	buildings12040417_makarova	0
+Their feedback would be valuable for designers to note that application of the biophilic design attributes in the office design can enhance the experiences and evaluations of workers.			Something I felt would useful would at the beginning would be a clear definition for what “biophilic attributes” refers to. Figure 1 provides some insights into this, but these listed attributes span across different domains and the communal features are not that apparent. This becomes an issue later, since some aspects referring to biophilic design become unclear.	1	2	buildings12040417_makarova	0
+The figures and pictures have been adjusted in the revised manuscript.			P4, L173-174: The authors state that seven of the patterns from the 14 patterns of biophilic design were discarded. If this was the case, then please better articulate its overarching utility in this study, considering that half of the patterns were not relevant to the research scope.	1	2	buildings12040417_makarova	0
+These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results).			P4, L180-183: In traditional POE studies and general building science research, daylight, thermal comfort, and air-quality, would be considered as indoor environment parameters (as examples, please see refs. in comment #4), while office layout and building form would be considered a physical and architectural parameters. Reading further to page 5, lines 189-192, the authors begin to suggest to this, but referred to them and others indoor environmental parameters as factors for the workplace. I would suggest better rationalising the connections between the nine design parameters to biophilia to make these more overt.	1	2	buildings12040417_makarova	0
+Biophilic design is certainty an area worthy of interest, and the authors’ endeavours along this research line were appreciated and were something, I felt, contributes the knowledge in this domain.			Please consider simplifying the figure. Figure 1: The image presenting all the linkages is very interesting and is worth emphasising, but contains an overwhelming degree of information, and the text and line sizes are too small for readership. For example, some text boxes many not need further explanation (e.g. presence of water); also the lines connecting column A to the same patterns in column 4 could be colour coordinated.	1	2	buildings12040417_makarova	0
+Step two, we neglect the patterns which are not representative in office environment (please find the detailed explanations in the respond for Comment #8) and specify the selected design patterns to nine biophilic design attributes.			Table 2: Please consider providing further explanations for this table. It was not clear what the authors wanted to show.	1	2	buildings12040417_makarova	0
+Response 9: We believe that there is no conflict between the different classifications.			P8, L245: Please specify why these two offices were of interest (e.g., were they comparable or had specify architectural features worthy of study). If possible, please provide more characteristics (e.g., size, floor area, furniture layout (e.g., open-plan or enclosed), etc.) for each office. Later (P10, L299), it says 201 questionnaires were collected, with 161 occupants taking part in the Singaporean office. This led me to believe that this office was much larger than the building studied in China. An image showing the indoor conditions and outdoor façade for each might be beneficial. Many of the explanations found in section 3.1 could be moved into this part of the manuscript, since they many describe and show the existing office conditions and to do necessarily form part of the main results.	1	2	buildings12040417_makarova	0
+While there is likely some share attributes (e.g.			Section 3.3. Although I appreciated the thoroughness to which the descriptive statistical was explained, I wasn’t convinced the mean was the best indicator for the data, considering that evaluation scores were collected on a 5-point scale and not a continuous linear one. In-lieu of the mean, please consider using the median and inter-quartile range as the central tendency and dispersion indicators. Figures 1 and 4 can be removed, as the assumption of normality no longer applies (also on P15, L399-400), or replaced with boxplots.	1	2	buildings12040417_makarova	0
+Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement).			Table 8: Please consider applying benchmarks for what constitute reasonable levels for internal consistency, when using the Cronbach’s Alpha (e.g., α>0.7): Please see, for example: Taber, 2018. The use of Cronbach’s Alpha when developing and reporting research instruments in science education. Research in Science Education. Tavakol et al. Making sense of Cronbach’s Alpha. International Journal of Medical Education.	1	2	buildings12040417_makarova	0
+POE scales do not likely target biophilic elements in architectural designs directly, since a direct question (e.g.			Figure 3. The plot is well presented. A few minor notes for improvement: 1) Please consider adding short or abbreviated labels referring to the actual question, instead of codes (e.g., GH3-Q10). This would make it easier for the reader; 2) Round the percentages to the nearest whole number.	1	2	buildings12040417_makarova	0
+This seems to be a core aspect of their work but did really emerge from the final section of their work in the same way it was emphasized in the abstract.			P17, L434: Please correct the unfortunate citation error on this line.	1	2	buildings12040417_makarova	0
+c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration.			P18, section 4.2: Similar to comment #13, the data may be more suited to a Spearman’s correlation test, instead of the Pearson’s correlation coefficient. Due to the reasonable size of the dataset collected, it may not change the interpretation, but would help improve the analytical rigour.	1	2	buildings12040417_makarova	0
+2, Photo in table 6 should check the copyright of these photo.			The sentence reads: Homogenous subsets with significant discrepancies (differences?) across subsets, leading to no significant differences across subsets. The above is not easy to grasp. If the information is accurate, please consider amending this to make this clearer. P18, L448-450: Please check whether the sentence is accurate, and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3.	1	2	buildings12040417_makarova	0
+1) these two cases are limited in representing all the workplace biophilic designs.			While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavours, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work, but did really emerge from the final section of their work in the same way it was emphasised in the abstract.	1	2	buildings12040417_makarova	0
+While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.			For their response to comment #6, while I generally agreed with the authors, I felt that it didn’t quite address what I tried to originally convey. POE scales do not likely target biophilic elements in architectural designs directly, since a direct question (e.g., how satisfied are you with the biophilic features) may not accurately depict every beneficial nuance they offer (e.g., psychological recovery). Mayer’s connectedness to nature scale) to help measure them. My generally feeling is that scales, from other domains (i.e., outside of POE studies) have been adopted for this reason, and a short sentence explaining the rationale supporting the lack of scales for biophilic design could be provided to briefly mentioned this. Instead studies may elect to use, for example, other relatable scales, which may not have originally been designed for POE surveys (e.g.	3	2	buildings12040417_makarova	0
+Naturalistic shapes and forms the shapes of plants on building facades and columns, animal facsimiles woven into fabrics and coverings.			If this was the case, perhaps this could be succinctly mentioned. In consideration to their response #8, I understood and agreed with why half the seven of the 14 papers were discarded, but it did not provide much insight into why the Patterns of Biophilic Design were still used. A more useful example, which may have elucidated this more, was whether are there too few frameworks available, and/or none others could have been used.	3	2	buildings12040417_makarova	0
+Author Response Response to Reviewer 1 Comments I would like to thank the authors for taking all my comments into full consideration.			"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_makarova	0
+These are not significant remarks, but nonetheless, might provide further improvements and clarity: #1: For their response to comment #6, while I generally agreed with the authors, I felt that it didn’t quite address what I tried to originally convey.			Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_makarova	0
+Kent, M., Parkinson, T., Kim, J., Schiavon, S., 2021.			The result of biophilic design frameworks for specific workplace typology is of certain value. The location of the workplace selected in this article is limited, the related biophilic factors are scientifically screened, and the researchers have made a detailed classification study. However, the huge research scope has certain obstacles to the relevant results, and the general and targeted conclusions will be worse.	1	2	buildings12040417_makarova	0
+The figures and pictures have been adjusted in the revised manuscript.			It is best to supplement and describe the necessity of research. At the same time, it will be more complete if the results are reflected in the summary.	1	2	buildings12040417_makarova	0
+There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace.			The object of the questionnaire should be composed of people with different ages and genders, which is best explained in the article.	1	2	buildings12040417_makarova	0
+Thermal & Air flow variability Subtle changes in air temperature, relative humidity, air ow across the skin, and surface temperatures that mimic natural environments.			Adding some explanations, examples and data comparison to the conclusion will be more intuitive and convincing, just as discussed earlier in the article. Perhaps this makes this article more complete and credible. At the same time, the conclusion only summarizes the article, and lasks discussions and explanations for the future research direction.	1	2	buildings12040417_makarova	0
+We think it is not hard for the readers to understand, and the original percentages (round to one decimal place) is more accurate.			The theoretical part of the presentation is extensive, but the application part is relatively poorly described.	1	2	buildings12040417_makarova	0
+Animals Re-presentation of nonhuman animal life 6.			It is better to supplement the temperature, humidity and other parameters of the selected office in the part of the experiment, so as to facilitate readers' reference rather than just giving the location.	1	2	buildings12040417_makarova	0
+Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020.			The author can think about the impact of such a pro biological design model on the psychology of different experimental personnel. I think psychological factors will also affect human physiological comfort.	1	2	buildings12040417_makarova	0
+#2: P1, L39-41: Please provide references to these frameworks.			Figure 1 should be improved regarding the arrow.	3	2	buildings12040417_makarova	0
+Instead, studies may elect to use, for example, other relatable scales, which may not have originally been designed for POE surveys (e.g.,			Photo in table 6 should check the copyright of these photo.	3	2	buildings12040417_makarova	0
+Yin, J., Arfaei, N., MacNaughton, P., Catalano, P.J., Allen, J.G., Spengler, J.D., 2019.			"Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."	3	2	buildings12040417_makarova	0
+Point 2: The illustrations in the article are small and a bit vague, some pictures can shrink a little, not to the top to the border, these can be optimized.			The definition of biophilia is described as an “inherent love” toward nature. While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.	1	2	buildings12040417_perova	0
+#3: P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment.			P1, L39-41: Please provide references to these frameworks. Reading further to page 3, I believe these are the 24 biophilic design attributes [ref. 25,39], and the 14 patterns of biophilic design [ref.40]. Further references around the biophilic concept could also be provided, e.g.: Bjørn et al., 2009. The latter reference reviewed many international standards that advocate nature and biophilic design for view and building spaces, with examples given to the Singapore context. This somewhat overlaps with my next comment.	1	2	buildings12040417_perova	0
+Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018.			P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment. WELL v2 has several features for Nature and Mind, and Biophilia – Parts I and II, with quantitative assessment methods provided. Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief. Other standards likely incorporate biophilic elements in building architecture and could be worth highlighting. The general issue raised by the authors do not necessarily imply a lack of guidelines for biophilic design, since there are several readily available, but may point toward prioritization or emphasis of criteria to meet certain varying expectations, which was alluded to on lines 52-53. If the authors agree with this, perhaps this could be revised here to reflect this.	1	2	buildings12040417_perova	0
+#11: Table 2: Please consider providing further explanations for this table.			P2, L63: Although I wouldn’t completely rule this out, POE surveys may not always provide feedback to the architect, since they are implemented post design-stage and the building would be operated by facility management or the owner. In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues. Recently POE studies, also using office data, advocate this as benefit to their implementation, albeit not necessarily being the only reason: žGraham et al., 2020. Building & Cities. Kent et al., 2021. Building and Environment. Cheung et al. 2021.	1	2	buildings12040417_perova	0
+Response 7: 1) The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing.			P3, L92: I think refers to “has helped” given the five decades predating this.	1	2	buildings12040417_perova	0
+Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being?			P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation. Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32. Therefore, it would be more appropriate to use psychological scales (e.g., PANAS or psychological restoration), instead of design orientated question or survey. If the authors agree with this, this aspect could be revised.	1	2	buildings12040417_perova	0
+When the occupants feel that they have a strong sense of relationship with nature, it is observed that the biophilic environment would have positive impacts on their health.			Something I felt would useful would at the beginning would be a clear definition for what “biophilic attributes” refers to. Figure 1 provides some insights into this, but these listed attributes span across different domains and the communal features are not that apparent. This becomes an issue later, since some aspects referring to biophilic design become unclear.	1	2	buildings12040417_perova	0
+Thermal & Air flow variability Subtle changes in air temperature, relative humidity, air ow across the skin, and surface temperatures that mimic natural environments.			P4, L173-174: The authors state that seven of the patterns from the 14 patterns of biophilic design were discarded. If this was the case, then please better articulate its overarching utility in this study, considering that half of the patterns were not relevant to the research scope.	1	2	buildings12040417_perova	0
+The theoretical part of the presentation is extensive, but the application part is relatively poorly described			P4, L180-183: In traditional POE studies and general building science research, daylight, thermal comfort, and air-quality, would be considered as indoor environment parameters (as examples, please see refs. in comment #4), while office layout and building form would be considered a physical and architectural parameters. Reading further to page 5, lines 189-192, the authors begin to suggest to this, but referred to them and others indoor environmental parameters as factors for the workplace. I would suggest better rationalizing the connections between the nine design parameters to biophilia to make these more overt.	1	2	buildings12040417_perova	0
+Limitation and Future Studies is added in the revised version: Revision in Page 23 Line 542-551: “6.			Figure 1: The image presenting all the linkages is very interesting and is worth emphasizing, but contains an overwhelming degree of information, and the text and line sizes are too small for readership. Please consider simplifying the figure. For example, some text boxes many do not need further explanation (e.g., presence of water); also, the lines connecting column A to the same patterns in column 4 could be color coordinated.	1	2	buildings12040417_perova	0
+Due to the reasonable size of the dataset collected, it may not change the interpretation, but would help improve the analytical rigour.			Table 2: Please consider providing further explanations for this table. It was not clear what the authors wanted to show.	1	2	buildings12040417_perova	0
+Dimension Office A Office B Location Singapore Shenzhen, South China Climate Zone Tropical Monsoon climate Sub-tropical climate Coordinate 1°16′North, 103°5′East 22°55′North, 114.1°East Floor 8 10 Office Ventilation Type Central air conditioned Natural ventilation Temperature in the office 25 to 26° C 26 to 28°C No.			P8, L245: Please specify why these two offices were of interest (e.g., were they comparable or had specify architectural features worthy of study). If possible, please provide more characteristics (e.g., size, floor area, furniture layout (e.g., open-plan or enclosed), etc.) for each office. Later (P10, L299), it says 201 questionnaires were collected, with 161 occupants taking part in the Singaporean office. This led me to believe that this office was much larger than the building studied in China. An image showing the indoor conditions and outdoor façade for each might be beneficial. Many of the explanations found in section 3.1 could be moved into this part of the manuscript, since they many describe and show the existing office conditions and to do necessarily form part of the main results.	1	2	buildings12040417_perova	0
+International Journal of Environmental Research and Public Health.			Section 3.3. Although I appreciated the thoroughness to which the descriptive statistical was explained, I wasn’t convinced the mean was the best indicator for the data, considering that evaluation scores were collected on a 5-point scale and not a continuous linear one. In-lieu of the mean, please consider using the median and inter-quartile range as the central tendency and dispersion indicators. Figures 1 and 4 can be removed, as the assumption of normality no longer applies (also on P15, L399-400), or replaced with boxplots.	1	2	buildings12040417_perova	0
+Revision in Page 13 Line 381-383: “The Cronbach’s α coefficient value of the main scale is 0.72, while those of the sub-scales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).” Citations are added in the Reference list: 57.			Table 8: Please consider applying benchmarks for what constitute reasonable levels for internal consistency, when using the Cronbach’s Alpha (e.g., α>0.7): Please see, for example: Taber, 2018. The use of Cronbach’s Alpha when developing and reporting research instruments in science education. Research in Science Education. Tavakol et al. Making sense of Cronbach’s Alpha. International Journal of Medical Education.	1	2	buildings12040417_perova	0
+A Data-Driven Analysis of Occupant Workspace Dissatisfaction.			Figure 3. The plot is well presented. A few minor notes for improvement: 1) Please consider adding short or abbreviated labels referring to the actual question, instead of codes (e.g., GH3-Q10). This would make it easier for the reader; 2) Round the percentages to the nearest whole number.	1	2	buildings12040417_perova	0
+Response 6: The different genders and different ages are included in the study.			P17, L434: Please correct the unfortunate citation error on this line.	1	2	buildings12040417_perova	0
+The above contents have been corrected in the revised version.			P18, section 4.2: Similar to comment #13, the data may be more suited to a Spearman’s correlation test, instead of the Pearson’s correlation coefficient. Due to the reasonable size of the dataset collected, it may not change the interpretation, but would help improve the analytical rigor.	1	2	buildings12040417_perova	0
+I think psychological factors will also affect human physiological comfort.			P18, L448-450: Please check whether the sentence is accurate and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3. The sentence reads: Homogenous subsets with significant discrepancies (differences?) across subsets, leading to no significant differences across subsets. The above is not easy to grasp. If the information is accurate, please consider amending this to make this clearer.	1	2	buildings12040417_perova	0
+Physiological and cognitive performance of exposure to biophilic indoor environment.			While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavors, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work but did really emerge from the final section of their work in the same way it was emphasized in the abstract.	1	2	buildings12040417_perova	0
+A Review of Psychological Literature on the Health and Wellbeing Benefits of Biophilic Design.			"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	0
+Biophilia, biophobia, and natural landscapes.			"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	0
+Response 7: The method of selection of the biophilic design attributes/patterns for workplace is: Step one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks.			"The introduction can be optimized appropriately.The introduction of the term ""Biophilia"" can be more detailed and easy to understand .By contrast, the introduction of “Biophilic Design”is very substantial.It may be better to find some literature to combine workplace design and “Biophilic Design”,and then to explore the relationship between them."	1	2	buildings12040417_perova	0
+The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE).			Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	0
+Although this I don’t believe this would be a large undertaking, the analysis would be better served by non-parametric tests due to the level of measurement used (i.e.			Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	0
+Apart from the design evaluation, POE is also one of the mainstream research methods that can effectively diagnosing operation problems.			Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	0
+Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design.			Some of the results in the illustrations are obvious and can be reduced to less elaboration. Overall work of the article is sufficient, the results is has the certain significance, but its limitation is obvious to all.	1	2	buildings12040417_perova	0
+Please see the revised contents in the updated manuscript below: Revision in Page 5 Line 191-197: “For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended because they are proofed that benefits health.			Figure 1 should be improved regarding the arrow.	3	2	buildings12040417_perova	0
+Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement) is reduced, because the illustrations are obvious in the figure.			Photo in table 6 should check the copyright of these photo.	3	2	buildings12040417_perova	0
+#6: P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation.			"Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."	3	2	buildings12040417_perova	0
+Response We are grateful to the reviewer for their favorable review and constructive input 1-			How is subtotal resection defined in chordoma (% of volume remaining?)	1	2	cancers14040966_makarova	0
+Thus it would be helpful for the authors to clarify this.			What means Pathology 0 in table 4?	1	2	cancers14040966_makarova	0
+Response We are grateful to the reviewer for their favorable review and constructive input 1-			Thus it would be helpful for the authors to clarify this. The AMIGO suite has great capabilities, but when it comes to intraoperative imaging, it is CT and MRI scans that are made, and then using Brain Lab and possibly other software, that information is processed and redisplayed including 3D reconstructions.	1	2	cancers14040966_makarova	0
+The authors are to be commended for pursing this approach in a small series of patients with difficult diagnoses.			This does not detract from the value of the technique, but it is important to clarify that what really is happening is a variety of preoperative imaging is imported and co-registered into the Brain Lab, and then intraoperative MR and occasionally CT data is again obtained and processed and analyzed.	1	2	cancers14040966_makarova	0
+There is not much to add and publication can be recommended.			It is therefore difficult to state that the results were clearly improved or to even compare to other series, without larger numbers and direct comparison or classification of individual tumor locations and characteristics in differing series.	1	2	cancers14040966_makarova	0
+All that is needed is easy intra-operative access to CT and MR imaging and the ability to import and reconstruct the data real time.			How is subtotal resection defined in chordoma (% of volume remaining?)	1	2	cancers14040966_perova	0
+What means Volume reduction 0 and 0% in Table 4?			What means Pathology 0 in table 4?	1	2	cancers14040966_perova	0
+With these caveats clarified in a modified version, the paper is valuable and could be published.			when one reads it initially it would appear that there are many different modalities available for intraoperative imaging. However, the PET scan does not appear to have actually been used for intraoperative repeat imaging to assess resection, and fluoroscopy and 3D reconstructions using Brain Lab software are standard fare in many operating rooms. Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions. The authors appear to really only use CT and MRI intraoperatively, with the other modalities imported preoperatively into the Brain Lab navigational system. 3D reconstructions of newly acquired data is a capability of the software, and is available in many places. Thus it would be helpful for the authors to clarify this. The AMIGO suite has great capabilities, but when it comes to intraoperative imaging, it is CT and MRI scans that are made, and then using Brain Lab and possibly other software, that information is processed and redisplayed including 3D reconstructions. Making this clear is important for others who either use the technique or who want to do so. All that is needed is easy intra-operative access to CT and MR imaging and the ability to import and reconstruct the data real time. That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or). This does not detract from the value of the technique, but it is important to clarify that what really is happening is a variety of preoperative imaging is imported and co-registered into the Brain Lab, and then intraoperative MR and occasionally CT data is again obtained and processed and analyzed.	1	2	cancers14040966_perova	0
+The AMIGO suite has great capabilities, but when it comes to intraoperative imaging, it is CT and MRI scans that are made, and then using Brain Lab and possibly other software, that information is processed and redisplayed including 3D reconstructions.			The results, while admirable, cannot easily be compared to other series, as the numbers are small and each case has its own complex series of characteristics that make it unique. It is therefore difficult to state that the results were clearly improved or to even compare to other series, without larger numbers and direct comparison or classification of individual tumor locations and characteristics in differing series.	1	2	cancers14040966_perova	0
+Minor Issues: [4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively.			Based on the cell line RNA and protein data the expression level of MACC1 is highly variable rather than being present or not. Is there a similar variability present among the tissue samples? Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?	1	2	cancers14071773_makarova	0
+[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion.			The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models. However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours. In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.	1	2	cancers14071773_makarova	0
+"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e."			It would be nice to know the selumetinib sensitivity of the two used cell lines. The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50mg/kg) are quite high. Please discuss how the applied concentrations are related to the clinicaly used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.	1	2	cancers14071773_makarova	0
+In this study, the authors have shown that the same is also true for gastric/esophageal cancer.			The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.	3	2	cancers14071773_makarova	0
+"should be after ""Expression was correlated with survival and morphological characteristics."""			"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."	3	2	cancers14071773_makarova	0
+[3] Throughout the paper, 10^x wrongly displays as 10x (e.g.			[1] Figure 4 is missing. In the pdf I reviewed, the caption is there, but the figure is not.	1	2	cancers14071773_makarova	0
+In the in vitro analysis they investigated MACC1 expression in five cell lines and generated a cell line with MACC1 overexpression and a MACC1 knock-down cell line.			[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249). Please fix this inconsistency.	1	2	cancers14071773_makarova	0
+The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.			"[3] Throughout the paper, 10^x wrongly displays as 10x (e.g. ""105"" instead of ""10^5"" in lines 152 and 170, ""106"" instead of ""10^6"" in line 217, ""X2"" instead of ""X^2"" in line 241, ""108"" instead of ""10^8"" in lines 353, 354, 365, 366, 367, 368, 370, 371)."	1	2	cancers14071773_makarova	0
+Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 2 Major Issues:			[4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively. If the difference is statistically significant, please briefly explain/speculate why.	1	2	cancers14071773_makarova	0
+[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).	1	2	cancers14071773_makarova	0
+"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."			[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.	1	2	cancers14071773_makarova	0
+The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50 mg/kg) are quite high.			[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).	1	2	cancers14071773_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e. specify the 95% confidence interval."	1	2	cancers14071773_makarova	0
+Answer: We thank the reviewer for this comment.			[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values). Same for lines 425/426.	1	2	cancers14071773_makarova	0
+Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 2 Major Issues:			"[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion. If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g. ""MACC1 is an independent negative prognostic marker in AGE/S patients."" should be after ""Expression was correlated with survival and morphological characteristics."" and before ""To analyze the role of MACC1 in vitro the cell lines FLO-1 and OE33 were lentivirally manipulated."" and so on)."	1	2	cancers14071773_makarova	0
+[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.			The authors have satisfactorily addressed my comments.	3	2	cancers14071773_makarova	0
+Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 1: 1.			Based on the cell line RNA and protein data the expression level of MACC1 is highly variable rather than being present or not. Is there a similar variability present among the tissue samples? Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?	1	2	cancers14071773_perova	0
+The results are interesting and may have clinical relevance.			The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models. However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours. In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.	1	2	cancers14071773_perova	0
+Please discuss how the applied concentrations are related to the clinically used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.			It would be nice to know the selumetinib sensitivity of the two used cell lines. The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50mg/kg) are quite high. Please discuss how the applied concentrations are related to the clinicaly used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.	1	2	cancers14071773_perova	0
+In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.			The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.	3	2	cancers14071773_perova	0
+It would be nice to know the selumetinib sensitivity of the two used cell lines.			"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."	3	2	cancers14071773_perova	0
+Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 1: 1.			[1] Figure 4 is missing. In the pdf I reviewed, the caption is there, but the figure is not.	1	2	cancers14071773_perova	0
+In the in vitro analysis they investigated MACC1 expression in five cell lines and generated a cell line with MACC1 overexpression and a MACC1 knock-down cell line.			[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249). Please fix this inconsistency.	1	2	cancers14071773_perova	0
+Please discuss how the applied concentrations are related to the clinically used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.			"[3] Throughout the paper, 10^x wrongly displays as 10x (e.g. ""105"" instead of ""10^5"" in lines 152 and 170, ""106"" instead of ""10^6"" in line 217, ""X2"" instead of ""X^2"" in line 241, ""108"" instead of ""10^8"" in lines 353, 354, 365, 366, 367, 368, 370, 371)."	1	2	cancers14071773_perova	0
+Previous studies in colorectal cancer have shown that MACC1 expression (which is associated with poor prognosis) induces cell migration in vitro and metastasis formation in vivo, and these effects can be inhibited by selumetinib.			[4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively. If the difference is statistically significant, please briefly explain/speculate why.	1	2	cancers14071773_perova	0
+In the in vitro analysis they investigated MACC1 expression in five cell lines and generated a cell line with MACC1 overexpression and a MACC1 knock-down cell line.			[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).	1	2	cancers14071773_perova	0
+Minor Issues: [4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively.			[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.	1	2	cancers14071773_perova	0
+I recommend the following edits before publication.			[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).	1	2	cancers14071773_perova	0
+Author Response Dear Professor Mok, We appreciate that you give us the possibility to revise our manuscript and we thank the reviewers for their valuable comments.			"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e. specify the 95% confidence interval."	1	2	cancers14071773_perova	0
+If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values). Same for lines 425/426.	1	2	cancers14071773_perova	0
+If the difference is statistically significant, please briefly explain/speculate why.			"[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion. If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g. ""MACC1 is an independent negative prognostic marker in AGE/S patients."" should be after ""Expression was correlated with survival and morphological characteristics."" and before ""To analyze the role of MACC1 in vitro the cell lines FLO-1 and OE33 were lentivirally manipulated."" and so on)."	1	2	cancers14071773_perova	0
+In this study, the authors have shown that the same is also true for gastric/esophageal cancer.			The authors have satisfactorily addressed my comments.	3	2	cancers14071773_perova	0
+This might be achieved by using the combination of so-called Tumor Treating fields (TTFields) with targeted drug, I have the following suggestions about this manuscript:			What is the rationale of using 150 kHz TTFields? TTFields at 100 kHz has also shown significant differences as cytotoxicity at p<0.01 and p<0.001 in HepG2 and Huh-7D12 cells which have around 60% and 50% of cell survival. Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).	1	2	cancers14122959_makarova	0
+Response 7: We thank the reviewer for this question.			What is the ‘n’ number of the samples and experiments in each group? Also mention the ‘n’ number in all the experiments involved for invitro and invivo.	1	2	cancers14122959_makarova	0
+In the animal study we have now added IHC examination of beclin-1 and of GRP78, a marker for ER stress, as described in results sub-section 3.4.			LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group. How can the authors correlate autophagy with apoptosis? It is not so trustworthy that the only expression of LC3B indicate the treated condition have increased autophagy in the tumor tissue. Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.	1	2	cancers14122959_makarova	0
+Response 6: We thank the reviewer for this comment.			Several data have shown very high error bars in each group (especially Fig Can the author provide the tumor images which were harvested from mice?	1	2	cancers14122959_makarova	0
+*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			In the IHC experiment for PARP, the authors have incubated with primary antibody only for 30 min? Is that timing enough to get protein expression? What source of secondary antibody was used? In addition to IHC, I suggest performing western blot using PARP antibody where the full length and cleaved bands are observed in the same blot.	1	2	cancers14122959_makarova	0
+Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			The figure number is mislabeled in Fig 4D-LC3 Immunofluorescence.	1	2	cancers14122959_makarova	0
+What are the p53 status and the apoptosis signaling pathway function in N1S1 cells?			Why there is no error bar in the control group of all bar graphs? Please include error bars and re-calculate the statistical analysis for all the data wherever missing.	1	2	cancers14122959_makarova	0
+The manuscript is an interesting work related to a potential new therapy of hepatocellular carcinoma (HCC) by using TTFields in combination with a TKI (Sorafenib).			Figure legends of Fig2D is missing. Fig C is repeated in the legend. Please label the figures appropriately. It is so frustrating to understand.	1	2	cancers14122959_makarova	0
+Point 8: Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated?			There is no data in Fig 4A. Whole data is missing but have explained in the result section and in figure legends.??	1	2	cancers14122959_makarova	0
+I thank the authors for their extensive work and comprehensive review of the manuscript.			Line 337 seems over statement since the data are not shown in the manuscript.	1	2	cancers14122959_makarova	0
+Overall, the authors believe that this research demonstrates potential for concomitant TTFields and sorafenib application in the treatment of HCC.			Typos: Line 111, ‘invitro’ spelling is not correct. Double ‘and’ in Line 342. Need language and grammar check.	1	2	cancers14122959_makarova	0
+*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			This is an interesting scientific study, as it concerns the issue of hepatocellular carcinoma, it also has a clinical aspect. The materials and methods section are elaborated in the details. Therefore, I expect further additional examination in the future. References contain mostly publications printed in the last 10 years. The authors indicated that TTFields had potential to be a new treatment option of hepatocellular carcinoma. The present study was examined in terms of autophagy and apoptosis in the antitumor effects of TTFields and sorafenib. However, the exact mechanism of the combination therapy induced cell death is not yet known as the activation of autophagy in the combination therapy was not increased.	1	2	cancers14122959_makarova	0
+Were the mice perfused before collecting the tumors?			In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells). A very important missing is not showing the actual number of rats included in the final analyses (the ones who successfully received therapy for more than 18 hours/day). This must be added.	1	2	cancers14122959_makarova	0
+ Point 7: Why there is no error bar in the control group of all bar graphs?			What are the p53 status and the apoptosis signaling pathway function in N1S1 cells? Maybe the cytotoxicity data will reveal a different better frequency.	1	2	cancers14122959_makarova	0
+Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			Another puzzling experiment is the schedule for the in vivo work. Since the authors missed to add Figure 4A for timeline, based on Methods section the rats were treated for 5 days with TTFields and or sorafenib and a day later the rays were sacrificed. While a short “acute” follow-up is welcome the most important experiment should allow the follow-up for much more days to indeed observe the effect of TTFields added to sorafenib. I could not find an explanation for not including a long term follow-up. In my opinion, this is a key therapeutically experiment which must be done and included in the study.	1	2	cancers14122959_makarova	0
+Response 4: We thank the reviewer for this question.			There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing. Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is total different?	1	2	cancers14122959_makarova	0
+*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	1	2	cancers14122959_makarova	0
+*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.	1	2	cancers14122959_makarova	0
+The revised manuscript is trying to be responsive to previous review comments.			The authors claim that cytotoxicity was measured “by cell counting using iCyt EC800 (Sony Biotechnology) 123 flow cytometer, and expressed as a percentage relative to the control.” Does this imply that they counted the live cells and plotted the final data as percentage of untreated controls (as figure 1 suggests). However, is this a real cytotoxicity or a cell growth inhibition? Did they measure the adherent cells after trypsinization. Were the cells from supernatant counted (where are probably the majority of dead cells)? There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.	1	2	cancers14122959_makarova	0
+Kaplan-Meyer survival curves are mandatory when toxic treatments are combined and compared.			Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated? Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic? Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group. Did the authors check changes in blood vessels density. Were the mice perfused before collecting the tumors?	1	2	cancers14122959_makarova	0
+Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			Was Mycoplasma testing done routinely? Were the cells checked also for authenticity?	1	2	cancers14122959_makarova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			Is there any explanation why the combination of TTFields and sorafenib did not induce a significant level of autophagy as compared to untreated animals which invalid the initial hypothesis that “concomitant application of sorafenib and TTFields may increase stress levels enough to tilt autophagy towards the cell death pathway”.	1	2	cancers14122959_makarova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			The statement “TTFields concomitant with sorafenib induced a significant increase in apoptosis’ in the abstract section is overstated. When compared with sorafenib alone there is practically no difference. Moreover, TTFields failed to increases apoptosis when added to sorafenib and compared to sorafenib alone in one out of two human cells line investigated	1	2	cancers14122959_makarova	0
+Response to Reviewer 4 Comments In general, the manuscript is well written and provides novel and interesting data illustrating the possibility to enhance the efficacy of sorafenib in the therapy of hepatocellular carcinoma (HCC).			Finally, adding to all the above questions, I found a very weak Discussion section which must be extended. Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”. This is not a correct statement. While for in vitro data, the authors have data, for in vivo they used only one frequency of 150 Hz. At least one different dose should have been studied for comparison since this is a completely different tumor environment than the in vitro one.	1	2	cancers14122959_makarova	0
+However, the limited experimental design and paucity of strong data ask for more experiments to proof the feasibility of this combination for treating HCC.			1) The authors demonstrate the efficacy of TTFields in vivo even when used as monotherapy. As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib. However, no differences in expression of LC3 marker were observed between these groups (treated with TTFields or with sorafenib)(as shown in Figure 4D). Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F. What is the mechanism illustrating higher efficiency of sorafenib against HCC?	1	2	cancers14122959_makarova	0
+ANOVA = analysis of variance; HCC = hepatocellular carcinoma; RMS = root mean square; SEM = standard error of the mean; TTFields = Tumor Treating Fields.			2) Despite the expression of cleaved PARP was very low in the tumors treated with TTFields or sorafenib alone ( as shown in IHC-images in Figure 4F), the authors declare about ~ 20% of positive cells, as show in the graphs below IHC-staining. Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.	1	2	cancers14122959_makarova	0
+Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			3) It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenin induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.	1	2	cancers14122959_makarova	0
+Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			4) Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib ( when compared to the cells treated with TTFiealds and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 ( for both HCC cell lines). This might be helpful and make the in vitro data more relavant with the data shown in vivo.	1	2	cancers14122959_makarova	0
+Majority of rodent study in cancer research lose the control group early but the treated groups are followed up for much longer to delineate indeed the efficacy vs. toxicity.			1) Figure 4A is missing.	1	2	cancers14122959_makarova	0
+Author Response Please see attachment Author Response File: Author Response.docx			2) the different HCC cell lines were used for in vitro and in vivo experiments, therfore making difficult to compare these data.	1	2	cancers14122959_makarova	0
+However, I feel that the experiments around this concept were not designed well enough to support their new findings.			What is the rationale of using 150 kHz TTFields? TTFields at 100 kHz has also shown significant differences as cytotoxicity at p<0.01 and p<0.001 in HepG2 and Huh-7D12 cells which have around 60% and 50% of cell survival. Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).	1	2	cancers14122959_perova	0
+How can the authors correlate autophagy with apoptosis?			What is the ‘n’ number of the samples and experiments in each group? Also mention the ‘n’ number in all the experiments involved for invitro and invivo.	1	2	cancers14122959_perova	0
+Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group. How can the authors correlate autophagy with apoptosis? It is not so trustworthy that the only expression of LC3B indicate the treated condition have increased autophagy in the tumor tissue. Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.	1	2	cancers14122959_perova	0
+Author Response Please see attachment Author Response File: Author Response.docx			What was the total weight of livers in all groups? Please update the images of whole liver showing tumors on it, or at least pictures of tumors which were removed after sacrifice, if applicable.	1	2	cancers14122959_perova	0
+The present study was examined in terms of autophagy and apoptosis in the antitumor effects of TTField and sorafenib.			In the IHC experiment for PARP, the authors have incubated with primary antibody only for 30 min? Is that timing enough to get protein expression? What source of secondary antibody was used? In addition to IHC, I suggest performing western blot using PARP antibody where the full length and cleaved bands are observed in the same blot.	1	2	cancers14122959_perova	0
+What is even more confusing is that in figure 4 for time points 6 and 24 hours, where, despite keeping the same GAPDH bands, the cleaved PARP data are changed.			The figure number is mislabeled in Fig 4D-LC3 Immunofluorescence.	1	2	cancers14122959_perova	0
+Therefore, I expect further additional examination in the future.			Why there is no error bar in the control group of all bar graphs? Please include error bars and re-calculate the statistical analysis for all the data wherever missing.	1	2	cancers14122959_perova	0
+While I do not know if the authors have the technology to perform TTFields in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells.			Figure legends of Fig2D is missing. Fig C is repeated in the legend. Please label the figures appropriately. It is so frustrating to understand.	1	2	cancers14122959_perova	0
+ Point 7: Why there is no error bar in the control group of all bar graphs?			There is no data in Fig 4A. Whole data is missing but have explained in the result section and in figure legends.??	1	2	cancers14122959_perova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			Line 337 seems over statement since the data are not shown in the manuscript.	1	2	cancers14122959_perova	0
+Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”.			Typos: Line 111, ‘invitro’ spelling is not correct. Double ‘and’ in Line 342. Need language and grammar check.	1	2	cancers14122959_perova	0
+Author Response Please see attachment Author Response File: Author Response.docx			In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells). A very important missing is not showing the actual number of rats included in the final analyses (the ones who successfully received therapy for more than 18 hours/day). This must be added.	1	2	cancers14122959_perova	0
+In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells. While I do not know if the authors have the technology to perform TTFileds in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells. This must be done and included for a better understanding of TTFieldds activity from in vitro to in vivo data. What are the p53 status and the apoptosis signaling pathway function in N1S1 cells? Maybe the cytotoxicity data will reveal a different better frequency.	1	2	cancers14122959_perova	0
+Also mention the ‘n’ number in all the experiments involved for invitro and invivo.			Another puzzling experiment is the schedule for the in vivo work. Since the authors missed to add Figure 4A for timeline, based on Methods section the rats were treated for 5 days with TTFields and or sorafenib and a day later the rays were sacrificed. While a short “acute” follow-up is welcome the most important experiment should allow the follow-up for much more days to indeed observe the effect of TTFields added to sorafenib. I could not find an explanation for not including a long term follow-up. In my opinion, this is a key therapeutically experiment which must be done and included in the study.	1	2	cancers14122959_perova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing. Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is total different?	1	2	cancers14122959_perova	0
+While a short “acute” follow-up is welcome the most important experiment should allow the follow-up for much more days to indeed observe the effect of TTFields added to sorafenib.			Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	1	2	cancers14122959_perova	0
+The rigor science recommends at least 2 independent experiments with at least 7 animals randomized per group.			Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.	1	2	cancers14122959_perova	0
+Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			The authors claim that cytotoxicity was measured “by cell counting using iCyt EC800 (Sony Biotechnology) 123 flow cytometer, and expressed as a percentage relative to the control.” Does this imply that they counted the live cells and plotted the final data as percentage of untreated controls (as figure 1 suggests). However, is this a real cytotoxicity or a cell growth inhibition? Did they measure the adherent cells after trypsinization. Were the cells from supernatant counted (where are probably the majority of dead cells)? There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.	1	2	cancers14122959_perova	0
+Response to Reviewer 1 Comments In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated? Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic? Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group. Did the authors check changes in blood vessels density. Were the mice perfused before collecting the tumors?	1	2	cancers14122959_perova	0
+Did the authors check changes in blood vessels density.			Was Mycoplasma testing done routinely? Were the cells checked also for authenticity?	1	2	cancers14122959_perova	0
+Response to Reviewer 2 Comments This study identified the efficacy of TTFields treatment for hepatocellular carcinoma and the combined effect of sorafenib in vitro and in vivo.			Is there any explanation why the combination of TTFields and sorafenib did not induce a significant level of autophagy as compared to untreated animals which invalid the initial hypothesis that “concomitant application of sorafenib and TTFields may increase stress levels enough to tilt autophagy towards the cell death pathway”.	1	2	cancers14122959_perova	0
+Response 3: We sincerely appreciate this well-taken comment.			The statement “TTFields concomitant with sorafenib induced a significant increase in apoptosis’ in the abstract section is overstated. When compared with sorafenib alone there is practically no difference. Moreover, TTFields failed to increases apoptosis when added to sorafenib and compared to sorafenib alone in one out of two human cells line investigated	1	2	cancers14122959_perova	0
+*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			Finally, adding to all the above questions, I found a very weak Discussion section which must be extended. Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”. This is not a correct statement. While for in vitro data, the authors have data, for in vivo they used only one frequency of 150 Hz. At least one different dose should have been studied for comparison since this is a completely different tumor environment than the in vitro one.	1	2	cancers14122959_perova	0
+Moreover, many bands belonging to all investigated proteins are truncated, fractured and I identified a lot of troubleshooting in bands due the presence of bubbling when running the blots.			1) The authors demonstrate the efficacy of TTFields in vivo even when used as monotherapy. As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib. However, no differences in expression of LC3 marker were observed between these groups (treated with TTFields or with sorafenib)(as shown in Figure 4D). Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F. What is the mechanism illustrating higher efficiency of sorafenib against HCC?	1	2	cancers14122959_perova	0
+However, the limited experimental design and paucity of strong data ask for more experiments to proof the feasibility of this combination for treating HCC.			2) Despite the expression of cleaved PARP was very low in the tumors treated with TTFields or sorafenib alone ( as shown in IHC-images in Figure 4F), the authors declare about ~ 20% of positive cells, as show in the graphs below IHC-staining. Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.	1	2	cancers14122959_perova	0
+Another weird observation is the new figures 3D, E, and F. Despite showing the same fold changes graphs in letter to reviewer and updated manuscript, the bands are different and all over the place in the mentioned files.			3) It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenin induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.	1	2	cancers14122959_perova	0
+Response: We thank the reviewer for the positive review.			4) Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib ( when compared to the cells treated with TTFiealds and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 ( for both HCC cell lines). This might be helpful and make the in vitro data more relavant with the data shown in vivo.	1	2	cancers14122959_perova	0
+Response 1: We thank the reviewer for this question.			1) Figure 4A is missing.	1	2	cancers14122959_perova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			2) the different HCC cell lines were used for in vitro and in vivo experiments, therfore making difficult to compare these data.	1	2	cancers14122959_perova	0
+Leaking splicing was also noted in two instances resulting in detection of the canonical transcript in 26.3% and 21.3% of transcripts, respectively.			Line 156 – the authors refer to “ad-hoc rules” that they have developed for consideration of the different coding transcripts associated with the same spliceogenic variant in variant interpretation and classification. Although a reference is provided so that the reader can look up what these ad-hoc rules are, it would also be helpful to briefly describe these in the current manuscript.	1	2	cancers14122960_makarova	0
+The transcript analysis results demonstrated splice complexity with several variants resulting in two or more transcript isoforms.			Lines 250-251 and 263-265– In these lines the authors discuss transcripts in which the encoded proteins lack some beta strands. The lack of these protein structures are used as evidence to support pathogenicity. It is not clear from the text how lack of these beta strands is predicted to impact protein function. Is there evidence from another source that these beta strands are critical to protein function and that their loss is deleterious (rather than resulting in normal or slightly reduced protein activity)?	1	2	cancers14122960_makarova	0
+Author Response - Thank you very much for the positive comments and the revision of our manuscript.			Lines 259-274 – This paragraph refers to evidence used in the variant interpretation of three aberrant transcripts that kept the open reading-frame (Δ(E2p3), Δ(E5) and ▼(E8p3)). However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names. As such, in order to correlate the discussion in this paragraph with the information in table 2, the reader also needs to cross reference Table 1 or 3. Incorporation of the transcript isoform names in table 2 would assist the reader in correlating this discussion of transcript isoforms with the corresponding evidence used to classify each of the variants.	1	2	cancers14122960_makarova	0
+Lines 250-251 and 263-265– In these lines the authors discuss transcripts in which the encoded proteins lack some beta strands.			Lines 259-274 – This paragraph refers to evidence used in the variant interpretation of three aberrant transcripts that kept the open reading-frame (Δ(E2p3), Δ(E5) and ▼(E8p3)). However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names. As such, in order to correlate the discussion in this paragraph with the information in table 2, the reader also needs to cross reference Table 1 or 3. Incorporation of the transcript isoform names in table 2 would assist the reader in correlating this discussion of transcript isoforms with the corresponding evidence used to classify each of the variants.	1	2	cancers14122960_makarova	0
+Although mini-gene assays sometimes is not reflective for the splicing of whole gene in a particuar locus but it is now is an well accepted preliminary screening assay.			Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively. The discussion of these variants in this section (and in other sections) does not address the significance of canonical transcripts in these cases. Leaky splice variants have been reported in various genes and sometimes can be associated with milder phenotypes (or no phenotypes), presumably because the canonical isoforms contribute to a “phenotypic rescue”. Is it known whether there is a threshold of RAD51C deficiency that is tolerated before associated cancer risks become increased?	1	2	cancers14122960_makarova	0
+However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names.			Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively. The discussion of these variants in this section (and in other sections) does not address the significance of canonical transcripts in these cases. Leaky splice variants have been reported in various genes and sometimes can be associated with milder phenotypes (or no phenotypes), presumably because the canonical isoforms contribute to a “phenotypic rescue”. Is it known whether there is a threshold of RAD51C deficiency that is tolerated before associated cancer risks become increased?	1	2	cancers14122960_makarova	0
+Although mini-gene assays sometimes is not reflective for the splicing of whole gene in a particuar locus but it is now is an well accepted preliminary screening assay.			in the Introduction in the sentence 61-69 the authors do not mention anything about BRCA1 (they mention MLH1 though); I would expect that as BRCA1 splice variants are deeply studied.	1	2	cancers14122960_makarova	0
+While in silico prediction tools can provide insights to possible impacts of nucleotide variants on splicing, these prediction tools do not replace the empirical evidence that the minigene assay can provide.			In the Materials and Methods, the section 2.6 is not actually a section that described some methodology. It appears to be more appropriate as results and maybe some information could be in the introduction.	1	2	cancers14122960_makarova	0
+Actually, not many publications report variants affecting splicing of RAD51C.			The acronyms need to be explained. Not all readers are familiar with PTC (premature termination codon) and NMD (nonsense mediated decay); as some variants are found PTC-NMD, this needs to be explained better. I would also stress a little more that FL is almost undetectable.	1	2	cancers14122960_makarova	0
+The authors used bioinformatic analysis to sort out variants that potentially affect splicing and then analyzed them using mini-gene assays.			The acronyms need to be explained. Not all readers are familiar with PTC (premature termination codon) and NMD (nonsense mediated decay); as some variants are found PTC-NMD, this needs to be explained better. I would also stress a little more that FL is almost undetectable.	1	2	cancers14122960_makarova	0
+This was a very interesting manuscript that was well written.			Splicing variants are called solely based on the size difference. It is good to look into Sanger sequence of splice variants after gel elution of the band and sequencing.	1	2	cancers14122960_makarova	0
+Response 5: We agree with the reviewers' suggestion that the manuscript needs mild English proofing.			This manuscript adds evidence that NR2F1 is a dormancy marker. In addition, it shows that NR2F1 is primarily expressed in CAFs, particularly in inflammatory CAFs.	1	2	cancers14122962_makarova	0
+Log-rank test was used for the analysis, and significant p values are shown in bold.			Given that dormant disseminated tumor cells (DTCs) highly express NR2F1 (Fluegel et al. Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs. It would be nice, if the authors would discuss this important point in more detail.	1	2	cancers14122962_makarova	0
+Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?			The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels. It would be interesting to see, how NR2F1 levels would change with endocrine treatment and/or endocrine resistance.	1	2	cancers14122962_makarova	0
+Survival analysis between high and low NR2F1 group in triple-negative breast cancer.			Breast cancer is a heterogenous disease. Different subtypes behave differently in many aspects. It would be great to see some subtype-specific data.	1	2	cancers14122962_makarova	0
+The OS of NR2F1 high triple- negative and ER-positive/HER2-negative subtypes were associated with better survival in GSE96058, and DSS of NR2F1 high HER2-positive subtypes was associated with worse survival in METABRIC.			The authors state “We demonstrated that the expression of NR2F1, RARB, and TGFB1 genes are higher in previously established dormant cells (D2OR murine breast cancer cells [44] compared to the proliferative cells (D2A1 cells) in both 2D and 3D cultures (Figure 1A, all p < 0.02).” This is not true for NR2F1 in 2D. Please correct.	1	2	cancers14122962_makarova	0
+We found that NR2F1 is most predominantly expressed in CAFs in the TME of primary breast cancer.			The authors present interesting findings that a tumor dormancy marker, NR2F1, is predominantly expressed in the inflammatory CAFs, and high expression of NR2F1 is associated with suppressed immune response and increased density of stromal cells. However, this reviewer has a few concerns that need to be addressed before accepting this article for publication.	1	2	cancers14122962_makarova	0
+In tumors with high Nottingham grade, NR2F1 expression was found to be lower.			This reviewer noticed that all the analysis was performed on the public data sets of bulk RNA-seq or single cell sequencing of primary breast tumors. Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?	1	2	cancers14122962_makarova	0
+Survival analysis between high and low NR2F1 group in triple-negative breast cancer.			Moreover, the authors need to provide some, at least minimum, evidence or clues that CAF-expressed NR2F1 is responsible for tumor dormancy regulation.	1	2	cancers14122962_makarova	0
+After multi-group comparison, baseline NR2F1 expression and expression in each cell type were compared in two groups by the one-way ANOVA test.			Line 170, “Figure 1E” should be “Figure 1D”.	1	2	cancers14122962_makarova	0
+However, this reviewer has a few concerns that need to be addressed before accepting this article for publication.			Given that dormant disseminated tumor cells (DTCs) highly express NR2F1 (Fluegel et al. Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs. It would be nice, if the authors would discuss this important point in more detail.	1	2	cancers14122962_perova	0
+Results: Breast cancer (BC) with high NR2F1 expression, enriched TGFβ signaling, multiple metastases, and stem cell- related pathways.			The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels. It would be interesting to see, how NR2F1 levels would change with endocrine treatment and/or endocrine resistance.	1	2	cancers14122962_perova	0
+Log-rank test was used for the analysis, and significant p values are shown in bold.			Breast cancer is a heterogenous disease. Different subtypes behave differently in many aspects. It would be great to see some subtype-specific data.	1	2	cancers14122962_perova	0
+Kaplan-Meier curves of OS, DSS, and DFS based on the high and low NR2F1 expression in ER-positive/HER2-negative breast cancer of three large cohorts.			The authors state “We demonstrated that the expression of NR2F1, RARB, and TGFB1 genes are higher in previously established dormant cells (D2OR murine breast cancer cells [44] compared to the proliferative cells (D2A1 cells) in both 2D and 3D cultures (Figure 1A, all p < 0.02).” This is not true for NR2F1 in 2D. Please correct.	1	2	cancers14122962_perova	0
+To this end, we believe that NR2F1 expression in the bulk tumor does not reflect the expression in the cancer cells, thus its value as a biomarker is in doubt.			This reviewer noticed that all the analysis was performed on the public data sets of bulk RNA-seq or single cell sequencing of primary breast tumors. Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?	1	2	cancers14122962_perova	0
+(A) Boxplots showing various scores based on high and low NR2F1 expression in HER2 positive breast cancer of TCGA; Intratumoral heterogeneity, homologous recombination deficiency (HRD), silent/non- silent mutation rate, SNV/Indel neoantigen, Interferon gamma response, fraction altered, tumor-infiltrating lymphocytes (TIL) fraction, and stromal fraction.			Moreover, the authors need to provide some, at least minimum, evidence or clues that CAF-expressed NR2F1 is responsible for tumor dormancy regulation.	1	2	cancers14122962_perova	0
+It would be great to see some subtype-specific data.			Line 170, “Figure 1E” should be “Figure 1D”.	1	2	cancers14122962_perova	0
+We deeply appreciate the comment raised by the reviewer.			Some FT-IR peaks shift with adsorption of Cr, and some not. Please add the discussion which peak shift is related to Cr, the authors can combine these with the discussion of the mechanism.	1	2	catal12030290_makarova	0
+In such form is rather difficult to make adequate comparison.			In Figure 3, what is the origin of the increase concentration of Al and S?	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			In the experiment, H2SO4 is added in the first step, what is the purpose? Will this bring pollution problem?	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			How about the Cr concentration in the polluted ground water? Is the products suitable for this real condition?	1	2	catal12030290_makarova	0
+Often, spectra with different atomic abundances of elements can be obtained even from the same sample.			The issue of regeneration has not been sufficiently worked out, but the authors themselves write about this, and it is not clear why NaOH was used for this.	1	2	catal12030290_makarova	0
+However, we regret that we were not able to investigate the BET analysis due to pandemic situation, which could definitely give us additional information.			It was also not clear to me how much hydrochloric acid was eventually added during the preparation of the adsorbent.	1	2	catal12030290_makarova	0
+Therefore, the authors are recommended to add the missing information (i.e.			The authors did not indicate how the “separation factor ‘RL’ is calculated and did not give a link to the equation for its calculation, and if the readers are not quite in the subject, then what kind of factor they do not understand.	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			One could also estimate the specific surface area and porosity. So, the paper can be published after revision.	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			The authors should write the complete terms of all abbreviations (including the instruments) before the first use in the abstract and main manuscript i.e. FT-IR and SEM in abstract section et al.	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			The authors should clearly explain the innovation and importance of their work on the introduction of the manuscript. They should justify the value of the work and compare their work with previously similar published papers.	1	2	catal12030290_makarova	0
+Please add the discussion which peak shift is related to Cr, the authors can combine these with the discussion of the mechanism.			Fig. 1 - for a more effective visual comparison, authors recommended to provide SEM images of the same scale. In such form is rather difficult to make adequate comparison.	1	2	catal12030290_makarova	0
+Synthesis of bioadsorbent and evaluation of its structure using SEM and FT-IR spectroscopy.			First of all, authors have to attach a EDX mapping images before/after sorption of chromium ions. And the authors are strongly recommended to add XPS spectra to this section of revised manuscript. The XPS method is much more sensitive and more accurately determines changes in the chemical composition of samples.	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			Why authors did not use XRD technique for sample characterization?	1	2	catal12030290_makarova	0
+They should justify the value of the work and compare their work with previously similar published papers.			The adsorption capacities of Al-GNSC adsorbents at different contact times have been provided. Which kinetics are right? Please add missing information about appropriate kinetic model in revised manuscript.	1	2	catal12030290_makarova	0
+There are some points which must be edited or clarified by providing additional information or comments:			It is rather difficult to make an adequate comparison of certain properties (catalysts or sorbents) with the already available results, since the concentration of the pollutant and the mass of the loaded sorbent vary in each experiment. Therefore, the authors are recommended to add the missing information (i.e. conditions for testing sorbents of Cr(VI) ions) to Table 3.	1	2	catal12030290_makarova	0
+A point-by-point response to the reviewer-3 comments is appended below for your convenience.			In order to confirm proposed mechanism of Cr(VI) adsorption (illustrated on the fig 6) Authors should provide data on adsorption capacity of pristine groundnut shell activated carbon (not modified with Al).	1	2	catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			The conclusion section should be elaborated and improved. The author should bring specific conclusions in accordance with obtained results.	1	2	catal12030290_makarova	0
+All the modifications are shown in yellow color in the revised manuscript.			Moderate English changes required	1	2	catal12030290_makarova	0
+A control challenged by bacterial supernatant without liposomes was added for each assay to verify the expected cytotoxic activity 2.			The authors used the term liposome nanotraps. The authors have used conventional liposome preparations. It is not clear why they are calling the preparations as the nanotraps.	1	2	cells11010166_perova	0
+However, given the poor characterization of SLS, its unavailability from commercial providers and its extremely tedious purification, the mechanistic details of the SLS neutralization will require an extensive a project of its own and are beyond the scope of the current study.			The authors have used bacterial culture supernatants to examine the cytotoxic effect on the THP-1, Jurkat, and Raji cell lines. Quantitation fo the the culture medium is required (by measuring the total protein content, or by providing some quantitative indicator). Volume of the culture supernatant is mentioned in the results. It is not quantitative, and will vary from batch to bath of the bacterial culture.	1	2	cells11010166_perova	0
+Carlsen J, Cömert C, Bross P, Palmfeldt J. Optimized High-Contrast Brightfield Microscopy Application for Noninvasive Proliferation Assays of Human Cell Cultures.			LDH-release assay of cytotoxicity or MTT assay of cell viability should have been used for measuring the cell death.	1	2	cells11010166_perova	0
+The authors used the term liposome nanotraps.			It is well-known that SLO binds to the cholesterol-containing lipid bilayer. Therefore, it is obvious that the cholesterol-containing liposomes would neutralise SLO present in the bacterial culture. Therefore, no new information is provided with these experiments.	1	2	cells11010166_perova	0
+Therefore, it is obvious that the cholesterol-containing liposomes would neutralize SLO present in the bacterial culture.			In my opinion, differential inhibition of the SLS activity in GGS supernatant by the liposome preparations is interesting, and may provide new insights, if explored in more detail.	1	2	cells11010166_perova	0
+From left to right: Stained protein ladder, GAS ATCC 19165, GAS 31009, GAS 50362, GGS ATCC 12394, GGS 5109.09, GGS 5804.			Although the sensitivity of immune cells to GAS or GGS supernatants is shown in the first section of the results, were any controls used in the neutralization assays with liposomes?	1	2	cells11010166_perova	0
+What is the reason for using different amounts of supernatant with cytolysins depending on its source or the type of cell line it is tested against in figures 2 and 3?			In the material and methods section it is stated that the cells survival assays start with the addition of a fixed volume of supernatant but it is not clear to me the criteria for reflecting different amounts of supernatant in Figures 2 and 3. It should be explained more clearly. What is the reason for using different amounts of supernatant with cytolysins depending on its source or the type of cell line it is tested against in figures 2 and 3? This should be explained. Furthermore, it should be justified why this difference does not influence the comparison of the results with the different cell lines (THP1, Jurkat, Raji).	1	2	cells11010166_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			Some errors in the legend of Figure 4, such as EM/PHYT/BHT, EM/PHYT is not presented in Figure 4.	1	2	cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			In Figure 4, the results is represented in three times or signal one or other times. Therefore, you must rewritten the legend of Figure 4.	1	2	cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			We recommend “galenic” can be changed to herbaceous, because galenic is not usually to see for non-professional fields or general cosmetics reader.	1	2	cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			Less a parenthesis at line 132.	1	2	cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			At line 160, “Figureure” should be “Figure.”	1	2	cosmetics2030248_makarova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			At line 161 mentioned Tonset DCS, but does not explain what this word mean.	1	2	cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			In Table II, Tonset DCS is a units of temperature, it should be used comma instead of decimal point.	1	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			At line 167, “donateis” is typically used to represent donate money, we proposed change to be “provide”, in additional, this sentence is not fluent at grammar.	1	2	cosmetics2030248_makarova	0
+Second Round of Evaluation Round 2: and Author Response			The sentence in line 168 is not fluent at grammar.	1	2	cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			At line 189, it dors (does) not have sample named EBL, it needs to be check again.	1	2	cosmetics2030248_makarova	0
+Second Round of Evaluation Round 2: and Author Response			Integration of all the above problems, this post Journal does not provide future applications of phytic acid in cosmetics and its title is mentioned in herbal lotion but the experiment did not use any herb emulsion formulation. For the antioxidant capacity of phytic acid only do the experiment of DPPH that has not more in-depth study. Lot of grammatical errors is found in sentences. So we suggested to big modifications, then returned for review.	1	2	cosmetics2030248_makarova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			From the experimental results that phytic acid will decrease the thermal stability of the emulsion, but at line 220 mention that phytic acid has the effect of lipid peroxidation and therefore can be added as an anti-oxidant in the emulsion. So we recommend authors to describe these two points much clearer.	3	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			The conclusions reported are very short and poor. More discussion of the presented results is needed.	1	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			The authors missed to report other authors’ studies which agree or disagree with their findings.	1	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			What is the output of this study? What are the implications in the use of PA in galenic/cosmetic formulations?	1	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			What are the suggestions of the authors about the use of PA in Cosmetics based on their results? And what the parameters and the tricks that the formulators should be aware of while using PA in their products?	1	2	cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Some errors in the legend of Figure 4, such as EM/PHYT/BHT, EM/PHYT is not presented in Figure 4.	1	2	cosmetics2030248_perova	0
+The article is well written, and the overall study was well planned and professionally conducted.			In Figure 4, the results is represented in three times or signal one or other times. Therefore, you must rewritten the legend of Figure 4.	1	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			We recommend “galenic” can be changed to herbaceous, because galenic is not usually to see for non-professional fields or general cosmetics reader.	1	2	cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			Less a parenthesis at line 132.	1	2	cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			At line 160, “Figureure” should be “Figure.”	1	2	cosmetics2030248_perova	0
+The article is well written, and the overall study was well planned and professionally conducted.			At line 161 mentioned Tonset DCS, but does not explain what this word mean.	1	2	cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			In Table II, Tonset DCS is a units of temperature, it should be used comma instead of decimal point.	1	2	cosmetics2030248_perova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			At line 167, “donateis” is typically used to represent donate money, we proposed change to be “provide”, in additional, this sentence is not fluent at grammar.	1	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			The sentence in line 168 is not fluent at grammar.	1	2	cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			At line 189, it dors (does) not have sample named EBL, it needs to be check again.	1	2	cosmetics2030248_perova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Integration of all the above problems, this post Journal does not provide future applications of phytic acid in cosmetics and its title is mentioned in herbal lotion but the experiment did not use any herb emulsion formulation. For the antioxidant capacity of phytic acid only do the experiment of DPPH that has not more in-depth study. Lot of grammatical errors is found in sentences. So we suggested to big modifications, then returned for review.	1	2	cosmetics2030248_perova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			From the experimental results that phytic acid will decrease the thermal stability of the emulsion, but at line 220 mention that phytic acid has the effect of lipid peroxidation and therefore can be added as an anti-oxidant in the emulsion. So we recommend authors to describe these two points much clearer.	3	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			The conclusions reported are very short and poor. More discussion of the presented results is needed.	1	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			The authors missed to report other authors’ studies which agree or disagree with their findings.	1	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			What is the output of this study? What are the implications in the use of PA in galenic/cosmetic formulations?	1	2	cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			What are the suggestions of the authors about the use of PA in Cosmetics based on their results? And what the parameters and the tricks that the formulators should be aware of while using PA in their products?	1	2	cosmetics2030248_perova	0
+While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			I don’t really doubt the empirical findings of the paper (which I see as support for the hypothesis in Japan and possibly in Korea), but I dislike the approach of spending a lot of space on various, often pointless, tests. There should be one detailed analysis based on adequate tests, and a discussion about the results. After all, government expenditure is a political decision.	1	2	economies3040150_perova	0
+References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			An import question is why are the series for real GDP per capita are taken from PWT 7.0? These series are constructed to be used in cross-country studies, and they are not suitable for the analyses in the paper. See the paper by Johnson et al. (2009). This is a major flaw of the paper and the choice should be defended by showing that using the ‘standard’ real GDP series’ would give the same results. Or the GDP series, downloaded from World Development Indicators (local currency, constant prices), should be used.	1	2	economies3040150_perova	0
+The suggestions are listed below in no particular order of importance: 1.			The dependency ratio has not been included mechanically in most papers, as it is this one. It is supposed to pick up changes in demand for government expenditure, such as schooling and pensions. The general idea is that these have increased government expenditure, both because per-capita schooling and pensions have expanded and because the dependency ratio has increased. In most of the countries studied, as is well-known, the dependency ratio declined during the period studied, while expenditure per person probably increased. Japan is most likely an exception; it should have entered the demographic transition much earlier than the other countries. There is thus no a priory no reason to expect the dependency ratio to play an important role in the Wagner hypothesis in general. It can thus be removed from the analysis, or moved to a footnote. If the author(s) wish to pursue the approach of testing other potentially relevant variables, see Shelton 2007.	1	2	economies3040150_perova	0
+More generally, we venture the highly subjective view that despite the immense mathematical sophistication of cointegration tests and their application in perhaps thousands of studies, it is not obvious how much additional substantive insight has been gained in the wide variety of contexts in which these tests have been used.”			Figure 1 tends to hide co-movements between the variables by mixing of scales. I suggest the graphs are re-done using two y-axes and preferably logs.	1	2	economies3040150_perova	0
+Round 2: My recommendation is to accept the paper for publication.			In Table 5 we can get information about cointegration from the R2 and the t-values. Only Japan shows any promise, in all other cases R2 are very low. I would say they are suspiciously low for time series data. Are thy correct?	1	2	economies3040150_perova	0
+While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			Gregory–Hansen is used when one fails to find cointegration with standard tests. There is no point in testing countries such as Japan and Korea, when a superior test shows that they are cointegrated.	1	2	economies3040150_perova	0
+Unit roots and the demand for cigarettes in Turkey: pitfalls and possibilities.			The use of the MTAR model is dubious because of the small sample, and it should not be used when cointegration has been established. It is unlikely that there is enough information in the data to capture asymmetric effects and the standard procedure is to use MTAR when Engle-Granger fails to show cointegration.	1	2	economies3040150_perova	0
+The suggestions are listed below in no particular order of importance: 1.			What is the alternative hypothesis of the Pedroni test, that there is cointegration in any country or or in all countries? This should be made clear.	1	2	economies3040150_perova	0
+Round 2: Author Response to Reviewer 2 Thank you.			The authors provide a very provocative conclusion (see below), but it is not supported by the analyses in the paper. The single-country analysis would provide the same results if done carefully, and the panel data analyses are dubious both because of the small cross-section (six) and the heterogeneity of the sample. I don’t mind the use panel analyses they can be presented for illustrative purposes, noting the heterogeneity. However, the time series analyses should be done correctly. Moreover, it is quite obvious that in simple bivariate cases, graphs can be very informative. However, the author(s) do not manage to provide any useful information about the dependency ratio in the ‘old fashioned’ analyses; they do only in Table 5 when (unknowingly) testing for cointegrating using OLS.	1	2	economies3040150_perova	0
+Round 2: My recommendation is to accept the paper for publication.			Despite Biehl’s (1998) insightful essay, almost all empirical research on the topic has interpreted the Wagnerian proposition As Ram (1998, Pp. 149-150) has explained, considerations of “nonstationarity” of the variables or “spuriousness” of the correlations or regression do not have much bearing on such tests.	1	2	economies3040150_perova	0
+While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			The x-axes in Figure 1 should have years.	3	2	economies3040150_perova	0
+Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			List of references is scant and limited. It is necessary to add more recent works on the subject with alternative ways of testing for Wagner’s Law. Suggestions include “Testing the Validity of Wagner’s Law in Bolivia: A Cointegration and Causality Analysis with Disaggregated Data” by A. Bojanic (2013); “Wagner’s Law and Italian Disaggregated Public Spending: Some Empirical Evidences” by C. Magazzino (2010); and “Panel Data, Cointegration, Causality and Wagner’s Law: Empirical Evidence from Chinese Provinces” by Narayan, P.K., et al. (2008).	1	2	economies3040150_perova	0
+References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			The paper only lists one way of testing for Wagner’s hypothesis (i.e., analyzing the relation between the share on GDP of government spending and real GDP per capita) when in fact there are many ways of looking at the hypothesis. The authors should include at least a couple of recent ways of testing this law, in addition to the traditional way of testing for it.	1	2	economies3040150_perova	0
+Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			The variables used do not have the same units, which may clog the analysis of the paper. Specifically, real GDP per capita is measured in 2005 dollars while the share of government consumption on GDP is measured in current prices. Either both should be in real terms or both in current prices, but not as is currently the case.	1	2	economies3040150_perova	0
+Unit roots and the demand for cigarettes in Turkey: pitfalls and possibilities.			The descriptive stats in Table 2 reflect stats for the whole period? If so, the table should explicitly state this fact.	1	2	economies3040150_perova	0
+At any rate, it is not obvious that such sophisticated and complex tests of cointegration provide any useful additional insight regarding the empirical status of the hypothesis.			Why are there 3 graphs for the Philippines in Figure 1? By the way, if the variables were measured in either current prices or utilizing the same base year, these graphs would likely be quite different (same goes for all statistical analysis, it is likely to vary when the two variables are measured the same way) 6.	1	2	economies3040150_perova	0
+Round 1: The paper analyzes the validity of Wagner’s law in six East Asian countries during the period 1960- 2008.			What is the point of the dependency rate? The authors should expand on the need to add this variable and what exactly is it trying to capture. In the same vein, what do the authors mean by “however, it seems that the fall in young-dependency ratio was greater than the rise in the old-dependency rate” on p. 4?	1	2	economies3040150_perova	0
+This makes the unconditional correlations uninteresting.			In Table 4, delete the “generated by SAS” note.	1	2	economies3040150_perova	0
+General comments The authors apply Latent Growth Curve Model (LGCM) approach to characterize the growth path of China’s agricultural labor productivity between 2000 and 2010 using a balanced panel data.			Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to. Are the annual agricultural output data are measured in their nominal value (at current price) or in real value (at constant price)? If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions? Labor productivity needs to be measured using real value of agricultural output. Otherwise, it may reflect the changes in prices instead of productivity. This issue needs to be properly addressed.	1	2	economies4030013_makarova	0
+Otherwise, it may reflect the changes in prices instead of productivity.			Measurement issue: The authors rely on the Latent Growth Curve Modeling technique to study the trend growth of China’s agricultural labor productivity. It is not clear, however, why the authors rely on a dataset with only 10-year period to discuss labor productivity trend growth issue instead of using a dataset with longer time period if data is available. In addition, the authors may want to justify their approach by briefly discussing the advantage or shortcomings of their method comparing to other approaches in identifying the growth trend of China’s agricultural labor productivity.	1	2	economies4030013_makarova	0
+Besides, the authors must define better the data how the agricultural labor productivity was calculated.			Please be more specific. Appendix Figure A2: The figure presents the chain-linked index of output and input. It is not clear whether the output is measured in real value (at constant price) or nominal value (at current price).	1	2	economies4030013_makarova	0
+We would like to thank the Reviewer to recommend us to develop this part.			One question that it is necessary to correct is the number of figures in the text.	1	2	economies4030013_makarova	0
+As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios.			Besides, the authors must define better the data how the agricultural labor productivity was calculated. The agricultural output is in current or constant prices? This could have strong implications in one case or another, especially in the first case. The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama. So, if the agricultural output is in current prices, the growing prices will affect deeply the results. It must be clearer.	1	2	economies4030013_makarova	0
+The authors may want to have some brief discussion on more recent literature regarding China’s productivity estimates and policy related issues.			On the other hand, in the same line of text (pg. 4 line 3), the authors define the denominator of the agricultural labor productivity as the employed labors in the first sector. What does this first sector mean? Is it a synonymous of agricultural sector? If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access. In this case, it would be the primary sector labor productivity. The authors should also clarify if this sector included the forest products and labor.	1	2	economies4030013_makarova	0
+- We would like to thank the reviewer’s scrutiny.			The introduction is welldone, but it could be improved in some points. I have missed a review of the literature. The measurement of the agricultural labor productivity is an important theme in the literature, not only in the agricultural economics, but also in the economics literature. For example, there are two paper published in two important journals in economics recently that the authors omitted. These papers could be reinforcement about the relevance of this measurement of agricultural productivity: Gollin D, Lagakos D, Waugh ME (2014) Agricultural productivity differences across countries. American Economic Review 104(5):165–170; Gollin D, Lagakos D, Waugh ME (2014) The agricultural productivity gap. Quarterly Journal Economics 129(2): 939–993.	1	2	economies4030013_makarova	0
+If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access.			Other thing to improve the introduction would be writing the main objective of the paper. I think, this clear objective could improve the introduction about the target that the authors follow in this paper. In this version, this is relatively diffuse.	1	2	economies4030013_makarova	0
+On the other hand, in the same line of text (pg.			The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence. The authors wrote about these concepts, for example pg. 6 lines 22-23 and pg. 9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.	1	2	economies4030013_makarova	0
+It is not clear, however, why the authors rely on a dataset with only 10-year period to discuss labor productivity trend growth issue instead of using a dataset with longer time period if data is available.			Maybe, the footnote 5 could go to the table 2’s note.	1	2	economies4030013_makarova	0
+On the other hand, in the same line of text (pg.			One of the most interesting issues of the paper is the political change in 2004. Perhaps, more detailed reasons of the reform would enrich the causes to promote it. Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.	1	2	economies4030013_makarova	0
+In the map of agricultural labor growth in Chinese region, the legend must include which is considered a high growth, medium and low growth of this variable.			As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios. Maybe, some simple correlations between the growths of agricultural labor productivity vs land productivity and land-labor ratio could enrich this or future analysis.	1	2	economies4030013_makarova	0
+Thanks to his/her remarks, the paper has improved under many important aspects.			Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to. Are the annual agricultural output data are measured in their nominal value (at current price) or in real value (at constant price)? If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions? Labor productivity needs to be measured using real value of agricultural output. Otherwise, it may reflect the changes in prices instead of productivity. This issue needs to be properly addressed.	1	2	economies4030013_perova	0
+The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			Measurement issue: The authors rely on the Latent Growth Curve Modeling technique to study the trend growth of China’s agricultural labor productivity. It is not clear, however, why the authors rely on a dataset with only 10-year period to discuss labor productivity trend growth issue instead of using a dataset with longer time period if data is available. In addition, the authors may want to justify their approach by briefly discussing the advantage or shortcomings of their method comparing to other approaches in identifying the growth trend of China’s agricultural labor productivity.	1	2	economies4030013_perova	0
+The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence.			Please be more specific. Appendix Figure A2: The figure presents the chain-linked index of output and input. It is not clear whether the output is measured in real value (at constant price) or nominal value (at current price).	1	2	economies4030013_perova	0
+Issues to improve the paper: The introduction is welldone, but it could be improved in some points.			One question that it is necessary to correct is the number of figures in the text.	1	2	economies4030013_perova	0
+On the other hand, in the same line of text (pg.			Besides, the authors must define better the data how the agricultural labor productivity was calculated. The agricultural output is in current or constant prices? This could have strong implications in one case or another, especially in the first case. The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama. So, if the agricultural output is in current prices, the growing prices will affect deeply the results. It must be clearer.	1	2	economies4030013_perova	0
+Figure 3: It is not clear what the axis labels represent in figure 3.			On the other hand, in the same line of text (pg. 4 line 3), the authors define the denominator of the agricultural labor productivity as the employed labors in the first sector. What does this first sector mean? Is it a synonymous of agricultural sector? If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access. In this case, it would be the primary sector labor productivity. The authors should also clarify if this sector included the forest products and labor.	1	2	economies4030013_perova	0
+It is correct to be published and the authors have made a strong effort to consider my comments and suggestions.			The introduction is welldone, but it could be improved in some points. I have missed a review of the literature. The measurement of the agricultural labor productivity is an important theme in the literature, not only in the agricultural economics, but also in the economics literature. For example, there are two paper published in two important journals in economics recently that the authors omitted. These papers could be reinforcement about the relevance of this measurement of agricultural productivity: Gollin D, Lagakos D, Waugh ME (2014) Agricultural productivity differences across countries. American Economic Review 104(5):165–170; Gollin D, Lagakos D, Waugh ME (2014) The agricultural productivity gap. Quarterly Journal Economics 129(2): 939–993.	1	2	economies4030013_perova	0
+Labor productivity needs to be measured using real value of agricultural output.			Other thing to improve the introduction would be writing the main objective of the paper. I think, this clear objective could improve the introduction about the target that the authors follow in this paper. In this version, this is relatively diffuse.	1	2	economies4030013_perova	0
+If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access.			The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence. The authors wrote about these concepts, for example pg. 6 lines 22-23 and pg. 9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.	1	2	economies4030013_perova	0
+In the map of agricultural labor growth in Chinese region, the legend must include which is considered a high growth, medium and low growth of this variable.			Maybe, the footnote 5 could go to the table 2’s note.	1	2	economies4030013_perova	0
+The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence.			One of the most interesting issues of the paper is the political change in 2004. Perhaps, more detailed reasons of the reform would enrich the causes to promote it. Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.	1	2	economies4030013_perova	0
+- We would like to thank the reviewer’s scrutiny.			As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios. Maybe, some simple correlations between the growths of agricultural labor productivity vs land productivity and land-labor ratio could enrich this or future analysis.	1	2	economies4030013_perova	0
+Author Response Please find the attached file.			Please add information on the number of additions for different transform types and sizes in original and proposed variants.	1	2	electronics11050760_makarova	0
+158075-158084, doi: 10.1109/ACCESS.2019.2950388.			It is better that they replace them with other keywords to increase the reach of the manuscript. Keywords : We suggest that the authors should replace keywords such as “video coding” and “transform” because these keywords are already found in the article title.	1	2	electronics11050760_makarova	0
+The names of the researchers must follow the style of the journal format.			Introduction: The authors should add the main contribution s briefly at the end of the introduction.	1	2	electronics11050760_makarova	0
+The double quotation should be omitted from the research titles in the list of references.			Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	1	2	electronics11050760_makarova	0
+<Reviewer 1> “The manuscript is well written, well organized, and clear.			There is a lot of recent research out there that can be used for comparison. As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.	1	2	electronics11050760_makarova	0
+Also, they should clarify the limitations of the proposed method.			Conclusions: Future directions should be added to the conclusion section.	1	2	electronics11050760_makarova	0
+There is a lot of recent research out there that can be used for comparison.			The names of the researchers must follow the style of the journal format.	1	2	electronics11050760_makarova	0
+There is a lot of recent research out there that can be used for comparison.			Some links do not work in the reference list like [22] … etc.	1	2	electronics11050760_makarova	0
+It can be easily computed by Equation (14) that the number of additions of the proposed method is smaller for all block sizes than those in VTM-8.2.”			This article requires extensive proofreading. Authors should check the entire article to remove all extensive mistakes (grammatical and typos) and to improve English writing quality.	1	2	electronics11050760_makarova	0
+• Figures and Tables: All figures and tables are shown before they are used in the text.			The authors should add the main contributions briefly at the end of the introduction. Introduction Section: This comment still requires a response.	3	2	electronics11050760_makarova	0
+Zhang, Z.; Zhao, X.; Li, X.; Li, L.; Luo, Y.; Liu, S.; Li, Z.			The authors should add a section where they discuss comparing their results with those of existing research. Also, they should clarify the limitations of the proposed method. Discussion Section: The authors did not respond accurately to this comment.	3	2	electronics11050760_makarova	0
+Zhao, X.; Chen, J.; Karczewicz, M.; Said, A.; Seregin, V. Joint Separable and Non-Separable Transforms for Next-Generation Video Coding, In IEEE Transactions on Image Processing, 2018, vol.			Figures and Tables: All figures and tables are shown before they are used in the text.	3	2	electronics11050760_makarova	0
+Inverse Transform Using Linearity Hyeonju Song*, and Yung-Lyul Lee* *Digital Media System Laboratory, Department of Computer Engineering, Sejong University, Seoul, Korea.			Additions also affect complexity. Please add information on the number of additions for different transform types and sizes in original and proposed variants.	1	2	electronics11050760_perova	0
+The authors should add the main contributions briefly at the end of the introduction.			Keywords: We suggest that the authors should replace keywords such as “video coding” and “transform” because these keywords are already found in the article title. It is better that they replace them with other keywords to increase the reach of the manuscript.	1	2	electronics11050760_perova	0
+IEEE Transactions on Multimedia, 2022, vol. 24, pp. 400-414, doi: 10.1109/TMM.2021.3052348.			Introduction Section:	1	2	electronics11050760_perova	0
+Koo, M.; Salehifar, M.; Lim, J.; Kim, S.-H. Low Frequency Non-Separable Transform (LFNST), 2019 Picture Coding Symposium (PCS), 2019, pp. 1-5, doi: 10.1109/PCS48520.2019.8954507.			Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	1	2	electronics11050760_perova	0
+• Introduction Section: This comment still requires a response.			Discussion Section: There is a lot of recent research out there that can be used for comparison. As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.	1	2	electronics11050760_perova	0
+Comment) Conclusions: Future directions should be added to the Conclusion Section.			Conclusions: Future directions should be added to the conclusion section.	1	2	electronics11050760_perova	0
+In answer to the specific comments: Comment) Keywords : We suggest that the authors should replace keywords such as “video coding” and “transform” because these keywords are already found in the article title.			References List: The list of references is recent, and all references are related to the research topic but it is not sufficient for this study. The names of the researchers must follow the style of the journal format. The double quotation should be omitted from the research titles in the list of references. Some search names in the reference list begin an uppercase letter for each word (such as [4], [5] ... etc.) and others use only an uppercase letter in the first word (such as [2], [9] … etc. ), authors should standardize style. All journal names should be italic. Some references do not contain enough information such as [16], [18] … etc. Some links do not work in the reference list like [22] … etc. The list of references requires an extensive check.	1	2	electronics11050760_perova	0
+As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.			The authors should add the main contributions briefly at the end of the introduction. Introduction Section: This comment still requires a response.	3	2	electronics11050760_perova	0
+The authors should add a section where they discuss comparing their results with those of existing research.			The authors should add a section where they discuss comparing their results with those of existing research. Also, they should clarify the limitations of the proposed method. Discussion Section: The authors did not respond accurately to this comment.	3	2	electronics11050760_perova	0
+In Proceedings of the 14th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Geneva, Swiss, 19-27 March 2019; 19.			Figures and Tables: All figures and tables are shown before they are used in the text.	3	2	electronics11050760_perova	0
+The appliances are prioritized based on their need and demand to achieve optimum energy utilization.			In the proposed system solar PV, wind generator and battery are the main generating system. As the proposed system is in isolated mode, how continuity of power supply be maintained?	1	2	en15031006_perova	0
+Simulation results are presented to show the effectiveness of the proposed method.			The quality of the figures needs to be improved.	1	2	en15031006_perova	0
+The appliances are prioritized based on their need and demand to achieve optimum energy utilization.			Equations 4, 9, and 12 include some corrections and need to be modified.	1	2	en15031006_perova	0
+Point 2: In the proposed system solar PV, wind generator and battery are the main generating system.			It could be better to include base load ratings in the results section (Fig: 10)	1	2	en15031006_perova	0
+The appliances are prioritized based on their need and demand to achieve optimum energy utilization.			Sizing of renewable energy sources and battery systems were not properly explored.	1	2	en15031006_perova	0
+Point 4: A comprehensive deeper literature review is necessary to address the research issue, Also authors need to provide a literature survey in an organized way.			Please discuss how consumers’ comfortability is marinated in the proposed method.	1	2	en15031006_perova	0
+Point 6: Sizing of renewable energy sources and battery systems were not properly explored.			Authors should avoid using the Subjective pronoun “ we” in academic works.	1	2	en15031006_perova	0
+Point 5: All the figures need to be improved.			In such a study an experimental case must be presented in a specified region.	1	2	en15031006_perova	0
+Simulation results are presented to show the effectiveness of the proposed method.			The authors should carefully distinguish the new contributions of their work from the new existing studies.for example: DOI: 10.1016/j.est.2020.101221	1	2	en15031006_perova	0
+Our objective is to schedule and shift the controllable appliances by estimating hourly power generation and considering consumer demand as a variable function.			A comprehensive deeper literature review is necessary to address the research issue. Also, authors need to provide a literature survey in an organized way.-	1	2	en15031006_perova	0
+In this paper, a demand-side management system for a renewable-based isolated residential community is proposed.			All the figures need to be improved.	1	2	en15031006_perova	0
+1703121)’ for publication in the Fermentation.			English language should be improved.	1	2	fermentation8050212_makarova	0
+Table 1: The number of decimal places of some constituents is excessive.			"Sometimes the document is different to follow due to English issues. Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology. ""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH. Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al. (2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”."	1	2	fermentation8050212_makarova	0
+Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al.			Explain what FRBE is. It appears for the first time on Page 1, Line 17; Response: Thanks for your comment.	1	2	fermentation8050212_makarova	0
+(2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”.			The hot water extraction conditions were tested before?	1	2	fermentation8050212_makarova	0
+"""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH."			What do the authors mean by 24hpf (Page 4, Line 148)?	1	2	fermentation8050212_makarova	0
+I ask that the authors specifically address each of my comments in their responses.			Explain what AO solution is. It appears on Page 4, Line 162; Response: Thanks for your comment.	1	2	fermentation8050212_makarova	0
+Page 3, Line 124: What do you mean “oscillated for 10 minutes”?			Explain the role of sodium humate.	1	2	fermentation8050212_makarova	0
+The “oscillated for 10 minutes” means “shaked for 10 minutes”.			Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.	1	2	fermentation8050212_makarova	0
+The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied.			Table 1: The number of decimal places of some constituents is excessive. Example: for polysaccharide content, I suggest writing 552+3 for RBE.	1	2	fermentation8050212_makarova	0
+Thus, in my opinion, the current study is on a topic of relevance and general interest to the journal's readers.			Why did the water-soluble phenolic acids contents increase after the fermentation?	1	2	fermentation8050212_makarova	0
+Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.			Page 9, Line 280: Is Fig. 3b showing different groups of LPO rate, or is Fig.3a?	1	2	fermentation8050212_makarova	0
+Thus, in my opinion, the current study is on a topic of relevance and general interest to the journal's readers.			Page 10, Lines 306 and 307: check the 73.85 and 77.00 values. They seem interchanged with each other.	1	2	fermentation8050212_makarova	0
+The application of the Duncan test should be justified because it is less sensitive in this kind of study.			Rewrite the first sentence in Conclusions.	1	2	fermentation8050212_makarova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			Page 8, Line 261: Change “FRB” to “FRBE”.	1	2	fermentation8050212_makarova	0
+It consists of performing a solid-state rice bran fermentation using a bacteria mix.			In the reference list, write the names of the microorganisms in italic. Some journal titles are registered in abbreviated form, and others have a full name. Standardize, taking into account the rules of the journal.	1	2	fermentation8050212_makarova	0
+"Page 3, Line 124: Change ""ethyl acetate reagents"" to "" ethyl acetate reagent""."			The main comment addressing the article improvement is the methodology applied for statistical analysis. The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied. Later one-factor analysis of variance should carry out accordingly. The application of the Duncan test should be justified because it is less sensitive in this kind of study.	1	2	fermentation8050212_makarova	0
+"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			Figures 2a, 3a and 4a include photos whose resolution should be improved.	1	2	fermentation8050212_makarova	0
+Chinese Journal of Animal Science 2014, 50(05),79-82.			please include a more specific section about the statistical methods.	1	2	fermentation8050212_perova	0
+3b showing different groups of LPO rate, or is Fig.3a?			"Sometimes the document is different to follow due to English issues. Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology. ""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH. Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al. (2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”."	1	2	fermentation8050212_perova	0
+Author Response Honored reviewers, Thank you for your contribution to reviewing work.			Explain what FRBE is. It appears for the first time on Page 1, Line 17; Response: Thanks for your comment.	1	2	fermentation8050212_perova	0
+The article is a fair enough approach, please include a more specific section about the statistical methods.			The hot water extraction conditions were tested before?	1	2	fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			What do the authors mean by 24hpf (Page 4, Line 148)?	1	2	fermentation8050212_perova	0
+(2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”.			Explain what AO solution is. It appears on Page 4, Line 162; Response: Thanks for your comment.	1	2	fermentation8050212_perova	0
+The application of the Duncan test should be justified because it is less sensitive in this kind of study.			Explain the role of sodium humate.	1	2	fermentation8050212_perova	0
+Page 3, Line 124: What do you mean “oscillated for 10 minutes”?			Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.	1	2	fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			Table 1: The number of decimal places of some constituents is excessive. Example: for polysaccharide content, I suggest writing 552+3 for RBE.	1	2	fermentation8050212_perova	0
+Page 3, Line 124: What do you mean “oscillated for 10 minutes”?			Why did the water-soluble phenolic acids contents increase after the fermentation?	1	2	fermentation8050212_perova	0
+Author Response Honored reviewers, Thank you for your contribution to reviewing work.			Page 9, Line 280: Is Fig. 3b showing different groups of LPO rate, or is Fig.3a?	1	2	fermentation8050212_perova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			Page 10, Lines 306 and 307: check the 73.85 and 77.00 values. They seem interchanged with each other.	1	2	fermentation8050212_perova	0
+"""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH."			Rewrite the first sentence in Conclusions.	1	2	fermentation8050212_perova	0
+Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.			Page 3, Line 124: What do you mean “oscillated for 10 minutes”? Is mixing for 10 minutes?	1	2	fermentation8050212_perova	0
+Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			Page 8, Line 261: Change “FRB” to “FRBE”.	1	2	fermentation8050212_perova	0
+"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			In the reference list, write the names of the microorganisms in italic. Some journal titles are registered in abbreviated form, and others have a full name. Standardize, taking into account the rules of the journal.	1	2	fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			The main comment addressing the article improvement is the methodology applied for statistical analysis. The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied. Later one-factor analysis of variance should carry out accordingly. The application of the Duncan test should be justified because it is less sensitive in this kind of study.	1	2	fermentation8050212_perova	0
+Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			Figures 2a, 3a and 4a include photos whose resolution should be improved.	1	2	fermentation8050212_perova	0
+Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			What equipment was used for the texture profile analysis?	1	2	gels8040212_makarova	0
+Response 1: It is our great pleasure to get your professional comments.			Describe the emulsion blank gel.	1	2	gels8040212_makarova	0
+The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			"What does the term ""wet gel"" mean?"	1	2	gels8040212_makarova	0
+Response 1: It is our great pleasure to get your positive comments.			Line 60: define WPI at first use.	1	2	gels8040212_makarova	0
+Response 1: It is our great pleasure to get your positive comments.			Line 84-88: This is a summary of what was carried out and not a discussion, please remove.	1	2	gels8040212_makarova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			Figure 2: Increase the font of the legend.	1	2	gels8040212_makarova	0
+One main request would be to make it clearer what was the formulations and conditions tested.			Line 166: English check.	1	2	gels8040212_makarova	0
+B1, B2, and B3 looks biphasic, was this the case?			Figure 3: rename using latin alphabet	1	2	gels8040212_makarova	0
+Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels.			Figure 4: Increase legend as it is not visible. B1, B2, and B3 looks biphasic, was this the case?	1	2	gels8040212_makarova	0
+This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate.	1	2	gels8040212_makarova	0
+Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			Line 474: What is meant by optimize condition?	1	2	gels8040212_makarova	0
+This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing? This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.	1	2	gels8040212_makarova	0
+B1, B2, and B3 looks biphasic, was this the case?			Line 496: remove respectively	1	2	gels8040212_makarova	0
+The manuscript is presented in a well-structured manner.			Line 499. Characteristics or characterization?	1	2	gels8040212_makarova	0
+Figure 2 has been modified accordingly to make the legend clearer.			Line 500: Define PDI at first use Line 507: Clearance height or gap`?	1	2	gels8040212_makarova	0
+Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			Line 507: Clearance height or gap`?	1	2	gels8040212_makarova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			Line 512: Was 1% found to be within the viscoelastic region?	1	2	gels8040212_makarova	0
+Figure 2 has been modified accordingly to make the legend clearer.			Line 516: rephase	1	2	gels8040212_makarova	0
+This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			Line 530: English check	1	2	gels8040212_makarova	0
+The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			What equipment was used for the texture profile analysis?	1	2	gels8040212_perova	0
+Thanks very much for your responsible and efficient work in the process of our manuscript.			Describe the emulsion blank gel.	1	2	gels8040212_perova	0
+The references cited are relevant and current, mostly published within the last three years.			"What does the term ""wet gel"" mean?"	1	2	gels8040212_perova	0
+This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			One main request would be to make it clearer what was the formulations and conditions tested.	1	2	gels8040212_perova	0
+The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			Line 60: define WPI at first use.	1	2	gels8040212_perova	0
+Author Response Point 1: This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			Line 84-88: This is a summary of what was carried out and not a discussion, please remove.	1	2	gels8040212_perova	0
+Thanks very much for your responsible and efficient work in the process of our manuscript.			Figure 1: Include in Fig.1c what was the the % of WPI used.	1	2	gels8040212_perova	0
+Response 1: It is our great pleasure to get your positive comments.			Figure 2: Increase the font of the legend.	1	2	gels8040212_perova	0
+Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			Line 166: English check.	1	2	gels8040212_perova	0
+The article is very well written and only requires few minor changes.			Figure 3: rename using latin alphabet	1	2	gels8040212_perova	0
+The references cited are relevant and current, mostly published within the last three years.			Figure 4: Increase legend as it is not visible. B1, B2, and B3 looks biphasic, was this the case?	1	2	gels8040212_perova	0
+Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate.	1	2	gels8040212_perova	0
+This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			Line 474: What is meant by optimize condition?	1	2	gels8040212_perova	0
+The manuscript is presented in a well-structured manner.The references cited are relevant and current, mostly published within the last three years.The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing? This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.	1	2	gels8040212_perova	0
+B1, B2, and B3 looks biphasic, was this the case?			Line 496: remove respectively	1	2	gels8040212_perova	0
+One main request would be to make it clearer what was the formulations and conditions tested.			Line 499. Characteristics or characterization?	1	2	gels8040212_perova	0
+The article is very well written and only requires few minor changes.			Line 500: Define PDI at first use	1	2	gels8040212_perova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			Line 507: Clearance height or gap`?	1	2	gels8040212_perova	0
+Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels.			Line 512: Was 1% found to be within the viscoelastic region?	1	2	gels8040212_perova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			Line 516: rephase	1	2	gels8040212_perova	0
+Line 512: Was 1% found to be within the viscoelastic region?			Line 530: English check	1	2	gels8040212_perova	0
+Severe drought restricts crop growth and sig- 38 nificantly reduces yield worldwide (Burke, Lobell et al. 2009), (Hu and Xiong 2014).			The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424. Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier. To have a meaningful comparison, the authors need to show the expression level of these two genes in the drought assay (Lines 400-417). As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing. In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal. This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.	1	2	genes13030404_makarova	0
+This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.			It is interesting to see that only MDA content increases in response to drought. The possible mechanism/reason should be included in the discussion.	1	2	genes13030404_makarova	0
+The manuscript is well written and the presented evidence supports the conclusions being made.			The discussion needs to be further expanded. Esp. RNA-Seq results need to be further discussed.	1	2	genes13030404_makarova	0
+RNA-Seq results need to be further discussed.			There should be a summary (concluding statement at the end). This manuscript ends very abruptly.	1	2	genes13030404_makarova	0
+Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier.			"Pg 1 line 45: the wording ""and so on"" is awkward and imprecise. Please rephrase and indicate what other processes are being referred to."	1	2	genes13030404_makarova	0
+Quantitative real-time PCR showed that the expression lev- 26 els of key genes in the lignin biosynthesis pathway were much lower in the MIM319 lines than in 27 the wild type.			Pg 2 line 67: don't assume that everyone reading the manuscript are aware of miRNA roles in plant stress physiology.	1	2	genes13030404_makarova	0
+"""sequester the normal expression"" is nonsensical."			Pg 3 Plasmid and sweet potato genetic transformation section: Include relevant protocols instead of just referring to another manuscript. Citing the original source is required but so are details needed to reproduce and evaluate the soundness of the manuscript.	1	2	genes13030404_makarova	0
+"""sequester the normal expression"" is nonsensical."			"Pg 6 line 239: I don't understand the point being made here. ""sequester the normal expression"" is nonsensical. Please clarify and rephrase."	1	2	genes13030404_makarova	0
+In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal.			The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424. Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier. To have a meaningful comparison, the authors need to show the expression level of these two genes in the drought assay (Lines 400-417). As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing. In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal. This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.	1	2	genes13030404_perova	0
+Blocking IbmiR319a 21 in transgenic sweet potato (MIM319) resulted in a slim and tender phenotype and greater sensitivity 22 to drought stress.			It is interesting to see that only MDA content increases in response to drought. The possible mechanism/reason should be included in the discussion.	1	2	genes13030404_perova	0
+We also found that the lignin content was reduced, which 25 led to increased brittleness in MIM319.			The discussion needs to be further expanded. Esp. RNA-Seq results need to be further discussed.	1	2	genes13030404_perova	0
+45 Citation: Lastname, F.; Lastname, F.; Lastname, F. Title.			There should be a summary (concluding statement at the end). This manuscript ends very abruptly.	1	2	genes13030404_perova	0
+16 Abstract: MicroRNA319 (miR319) plays a key role in plant growth, development, and multiple re- 17 sistance by repressing the expression of targeted TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) 18 genes.			"Pg 1 line 45: the wording ""and so on"" is awkward and imprecise. Please rephrase and indicate what other processes are being referred to."	1	2	genes13030404_perova	0
+Generally, the 40 responses of plants to abiotic stress are similar, especially in the first phase—a rapid, os- 41 motic phase that inhibits shoot growth (Zhou, Li et al. 2014).			Pg 2 line 67: don't assume that everyone reading the manuscript are aware of miRNA roles in plant stress physiology.	1	2	genes13030404_perova	0
+It is 39 therefore essential that crops with drought tolerance traits are produced.			Pg 3 Plasmid and sweet potato genetic transformation section: Include relevant protocols instead of just referring to another manuscript. Citing the original source is required but so are details needed to reproduce and evaluate the soundness of the manuscript. The same applies to other parts of the materials and methods:	1	2	genes13030404_perova	0
+In our experience, we observed completely different responses when we tried to compare PEG, air drying, and water withdrawal.			"Pg 6 line 239: I don't understand the point being made here. ""sequester the normal expression"" is nonsensical. Please clarify and rephrase."	1	2	genes13030404_perova	0
+To have a meaningful comparison, the authors need to show the expression level of these two genes in the drought assay (Lines 400-417).			Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	genes13030404_perova	0
+At this scale, the reader can’t see much detail.			Figures. Figure 1: Are colors in Figure 1 A related to the hydrostratigraphy or depicting topography? If the latter applies, make sure to provide a scale.	1	2	geosciences12010037_makarova	0
+one showing unfiltered + simplified drainage network, and one showing faults).done -How the authors arrive at the simplified drainage network is not very clear			Figure 2: The left spine (or border) does not appear on the pdf reviewed. This figure could be improved by adding shaded satellite image to help the reader compare the mapped drainage systems (which were automatically mapped) and fault traces/lineaments with the real features observed.	1	2	geosciences12010037_makarova	0
+In this process, minor mismatches of the automated simplified lines were removed and small drainage segments were connected along major lineaments for further simplification and reduction of the dataset.			Figure 3: The text (lines 258-259) mentions magnetic lineaments as prominent features, which can be caused by faulting. Such features are hard to visualize. It would certainly help if they were highlighted directly on the figure.	1	2	geosciences12010037_makarova	0
+This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control.			Figure 4: This figure could go in the appendix. It is good to let the reader know that not all seismic surveys provided the same quality, but this is more a technical point.	1	2	geosciences12010037_makarova	0
+Results appear to be sound and supported by the illustrations.			Figure 7: What is special about the locations highlighted by the markers (crosses on Figure 7B and square markers on Figure 7C). At this scale, the reader can’t see much detail.	1	2	geosciences12010037_makarova	0
+Magnetic data and 2D seismic reflection profiles were used to image structural features, and fluid/gas leakage pathways.			Lines 271-272: Why is the signal quality so variable, and how were the categories (excellent, average and poor) defined?	1	2	geosciences12010037_makarova	0
+Figure 2 -Poor quality figure (low resolution) in the version available to the reviewer.			Lines 323-324: The authors should explain why low magnitude local seismic signals can be believed to represent permeable pathways promoting water or gas migration.	1	2	geosciences12010037_makarova	0
+Reviewer 1: The authors present an interesting case study integrating geophysical, geological and hydrochemical data to map potential pathways for water and gases across complex sedimentary formations in the Northern Territories, in Australia.			Line 360: Here and throughout the text, the word ‘recent’ is used to describe some tectonic activity. It would be good define what recent means in the context of this study. An indication is provided in the Discussion (Line 414), but it would go to state this value upfront in the text.	1	2	geosciences12010037_makarova	0
+The map differentiates between “BaseCambrian seismic faults”, Post-Wilton “Strike-slip” and “Reverse” faults, and “FAULTS”.			Discussion: One general comment, could the use of other tracers (radon, of stable isotopes of water) yield also insight on the origin of the fluids sampled in this study?	1	2	geosciences12010037_makarova	0
+What is included in the “Drainage and structural lineament analysis” plot?			Aeromagnetic survey data acquired over the last 50+ years were employed for mapping structural and lithological features. Although the aeromagnetic data is said to be employed for structural interpretation, details somewhat sketchy. Reference is made to “…prominent […] magnetic lineaments which can be caused by faulting”, and “…several circular low magnetization anomalies, approximately two kilometers in diameter, aligned with northwest trending lineaments”. It would be nice to include a map with these features and lineaments interpreted from the aeromagnetic datasets. This would also tie the observations based on this dataset closer to the seismic interpretation (see below).	1	2	geosciences12010037_makarova	0
+A misinterpretation can led to the targeting of circulation pathways that are in fact only artefacts.			Seismic reflection 2D lines (1989-2015). 8500 km. Interpretation of shallow horizons and faults using all publicly available geological and geophysical data of the Beetaloo region. Fault polygons defined for five horizons. Depth conversion using check-shot velocities from 26 wells. See comments to Figure 4. The depth maps provided in Figure 5 should be supplemented by isochore maps for the rock volumes between pairs of mapped reflectors. See also comments to Figure 7.	1	2	geosciences12010037_makarova	0
+Magnetic data and 2D seismic reflection profiles were used to image structural features, and fluid/gas leakage pathways.			Helium measurements from groundwater samples. With respect to the helium measurements, no detailed documentation of where these samples were collected is provided beyond referring to Geological and Bioregional Assessment Program (2021a) Fact sheet 12, and Geological and Bioregional Assessment Program (2021b) Regional tracer results from the Cambrian Limestone Aquifer. These are both summary documents. Please provide a reference to the primary documentation.	1	2	geosciences12010037_makarova	0
+According to the new caption shows the unfiltered surface drainage network.			Fault and surface drainage network mapping (tectonic geomorphology). The drainage network was extracted automatically. It is not clear if the “simplified” drainage network shown in Figure 2 was also extracted automatically. The data is summarized in Figure 8 (see also comments to Figure 8 below). It is not clear if the surface drainage network line dataset is based on the “unfiltered” or “simplified” data shown in Figure 2. Since n=130, one can assume the latter. It is however not made clear how this “simplified” network was generated, but there appear to be a substantial number of instances where the “simplification” provides apparent mismatches with the original data (see examples included in comments to Figure 2 below). The accuracy of the resulting plot can therefore be questioned. If the “simplified” network was generated automatically, I encourage the authors to do a manual QC of the results.	1	2	geosciences12010037_makarova	0
+what differentiates “FAULT” from a “BaseCambrianSeismic fault”?).			Line 303: “We reviewed the seismic data in detail at the intersections with north-northwest trending lineaments observed on the magnetic dataset (Figure 4).” No such lineaments are shown in Figure 4.	1	2	geosciences12010037_makarova	0
+If the primary documentation is not accessible, the authors should state this clearly.			Line 582. Link for supplementary materials does not work.	1	2	geosciences12010037_makarova	0
+Thank you, the details comments on those figures have been followed and new figures are provided Helium measurements from groundwater samples.			- Poor visual discretization of main surface fault traces and Post Wilton geophysical faults (maybe use different colour for the two?)	1	2	geosciences12010037_makarova	0
+However, there are still a couple of minor issues.			Figure 1B - Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin. Although this is described in the caption, it is not easy to grasp for someone not familiar with the stratigraphy in the area without spending time some time with paper and pencil.	1	2	geosciences12010037_makarova	0
+Please use contrasting colours to differentiate them.			Figure 2 - Poor quality figure (low resolution) in the version available to the reviewer.	1	2	geosciences12010037_makarova	0
+- Line352: “However, the elevated helium concentrations show no systematic pattern of increase with flow distance (as expected for in-situ production along the flow path) but a rather patchy regional pattern (Figure 8).” This should refer to Figure 9.done -Line 582.			- Very, very overloaded figure, screen-dump from a mapping programme?	1	2	geosciences12010037_makarova	0
+done Figure 1B -Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin.			- How the authors arrive at the simplified drainage network is not very clear	1	2	geosciences12010037_makarova	0
+Please find attached the revised manuscript with track changes as well as the reply to the detailed comments, directly in purple.			Figure 4 - Poor quality figure (low resolution) in the version available to the reviewer.	1	2	geosciences12010037_makarova	0
+Although the authors point out that more data is needed to identify the source of the gas, the observations made in this study confirm the presence of fault-controlled fluid flow pathways connecting the stratigraphic levels where the unconventional plays are located with shallower aquifers the Beetaloo basin.			-Many well-name labels are hard difficult to read/illegible. Consider replacing well names on the map with numbers at the well position (white circles, black typeface) and add a table in the legend listing number and corresponding name of the well.	1	2	geosciences12010037_makarova	0
+done Figure 4 -Poor quality figure (low resolution) in the version available to the reviewer.			-Thin grey seismic lines are not explained in the legend or caption. Why are these not classified in terms of signal quality?	1	2	geosciences12010037_makarova	0
+Alternatively the position of Figure 6 A, B and C should be indicated in Figure 4 as well as Figure 3. done Figure 5.			Consider adding isochore maps for intervals between interpreted reflectors. These often highlight tectonic accommodation space creation better than maps.	1	2	geosciences12010037_makarova	0
+The replies to detailed comments are in purples, directly below each comment Frery et al.			d) Explain/improve labels (what do you mean by “ntspr_2M_gw”? “Coast_10million”? “frameworkboundaries”? “State_Borders_10million”. Amend labels or explain in caption.	1	2	geosciences12010037_makarova	0
+One of the key issues in the Beetaloo Sub-basin, the risk of groundwater contamination from fracking-based production of the unconventional plays.			e) Several items in the legend appear to have the same or very similar signatures.	1	2	geosciences12010037_makarova	0
+modified The link between interpreted faults and fluid or gas escape features is a key feature of this figure.			Clean up map: Not possible to differentiate post-Wilton fault types properly in the figure.	1	2	geosciences12010037_makarova	0
+Consider removing labels not referred to in the text.			Could not find “Coast_10million”, “frameworkboundaries” or “State_Borders_10million” on the map. If these items are present, please use a more contrasting signatures to make it more visible. If they are not present on the map, remove these items from the legend.	1	2	geosciences12010037_makarova	0
+• Lines 271-272: Why is the signal quality so variable, and how were the categories (excellent, average and poor) defined?			Consider replacing the purple outline of the eastern and western sub-basin with light grey shading.	1	2	geosciences12010037_makarova	0
+Consider using higher contrast colours or thicker lines to highlight different faults on the map.			Consider using the most prominent colour (red) for the possible fluid escape features as this is the key element of the map. Bright green might also bee an option.	1	2	geosciences12010037_makarova	0
+It is not clear if the surface drainage network line dataset is based on the “unfiltered” or “simplified” data shown in Figure 2.			Not possible to differentiate “ntspr_2M_gw” (whatever that is) and “PossibleFluidEscapeStructures” (same colour).	1	2	geosciences12010037_makarova	0
+It is not clear if the surface drainage network line dataset is based on the “unfiltered” or “simplified” data shown in Figure 2.			The link between interpreted faults and fluid or gas escape features is a key feature of this figure. The map differentiates between “BaseCambrian seismic faults”, Post-Wilton “Strike-slip” and “Reverse” faults, and “FAULTS”. The categorization is not adequately explained (e.g. what differentiates “FAULT” from a “BaseCambrianSeismic fault”?).	1	2	geosciences12010037_makarova	0
+Including these is maybe not critical, but as mentioned in the initial review, thickness maps sometimes highlight structural lineaments quite well.			Consider adding a simple conceptual sketch highlighting the stratigraphic position of the different faults (and possible fluid escape structures– this would also help visualizing which stratigraphic intervals the potential fluid escapes originate from and which stratigraphic intervals are potentially connected along potential fault-related fluid flow pathways.	1	2	geosciences12010037_makarova	0
+This fact stresses the need for clarity when presenting data and results.			Please provide a more informative caption to what these plots show. What is included in the “Drainage and structural lineament analysis” plot?	1	2	geosciences12010037_makarova	0
+Not possible to differentiate “ntspr_2M_gw” (whatever that is) and “PossibleFluidEscapeStructures” (same colour).			Figure 9. Poor resolution in the copy provided for the review. Consider using higher contrast colours or thicker lines to highlight different faults on the map.	1	2	geosciences12010037_makarova	0
+This would also tie the observations based on this dataset closer to the seismic interpretation (see below).			With respect to the helium measurements, the references to the primary documentation requested by the reviewer in comments to v.1 of the manuscript are not provided, nor do the authors provide any comment as to why they deem this unnecessary. Section 4.3 Helium concentrations and isotopic composition measured in groundwater, is framed around results from previous studies. Providing references to these original studies (including access to the actual data that allow objective verification) rather than referencing to summary papers would be more in line with scientific publishing practice. If the primary documentation is not accessible, the authors should state this clearly.	3	2	geosciences12010037_makarova	0
+Consider replacing well names on the map with numbers at the well position (white circles, black typeface) and add a table in the legend listing number and corresponding name of the well.			Figure 4. Seismic reflection data quality is ranked and colour-coded as “Excellent”, “Average” and “Poor”. Where does the fourth category, “Fair”, which also is a qualitative category, fit in, and why not include this category in the legend?	3	2	geosciences12010037_makarova	0
+Development of the petroleum resources in this region is controversial and faces opposition from Traditional Owners as well as climate scientists, and environmentalists.			Figure 5. Isochron maps have been included in the supplementary material, the initial review suggested isoCHORE maps. Including these is maybe not critical, but as mentioned in the initial review, thickness maps sometimes highlight structural lineaments quite well. I don't know if this is the case here, but if they do, including isochore maps would enhance the readers' understanding of basin geometry and -infill.	3	2	geosciences12010037_makarova	0
+We followed those advises and updated the document consequently.			If I understand the map correctly, fluid escape features on the map are shown as red dots. Why does the legend show these as thick red lines?	3	2	geosciences12010037_makarova	0
+The approach used by the authors is innovative and allows to elaborate several hypotheses regarding the occurrence and movement of both paleo and more recent fluids.			Figure 8. Please keep the number of measurements included in each plot. The plot “Surface drainage network from this study” is identical to the original figure and should have n=130 measurements. According to the new caption shows the unfiltered surface drainage network. To my understanding this should correspond to Figure 2 a, which shows substantially more than 130 lineaments. How did you pick the ones you measured? Please clarify.	3	2	geosciences12010037_makarova	0
+in Sode et al CMAJ 2013 doi: 10.1503/cmaj.121636.			In the Abstract you state that “two patients during their course of Rivaroxaban presented severe hematoma”, but in Table 1 it appears that the two hematomas occurred in the enoxaparin group? Please clarify.	1	2	healthcare10030476_makarova	0
+It is still not clear how thrombophilia was defined.			The introduction is not well written. The structure is unclear, and the authors repeat themselves many times. For example, the section begins with introducing inherited thrombophilias and VTE risk in these conditions, then proceeds to surgery, then back again to thrombophilia where it repeats itself and even states other numbers for prevalences and VTE risks than previously stated. Furthermore, suitable references are lacking in some places. For the present study to be acceptable for publication, the introduction should be thoroughly revised.	1	2	healthcare10030476_makarova	0
+C) Page 2, lines 87-88: “In this regard it is widely demonstrated that almost half of surgical patients who developed VTE have a mild thrombophilic syndrome [18].” This is rather an astonishing statement, and as your source, you cite a paper from the ��Australian Family Physician”.			The Discussion section could benefit from a language revision, especially in the latter part, to make the flow better. Also, a brief discussion on the limitations of your study should be included.	1	2	healthcare10030476_makarova	0
+Authors, however, need to address the following concerns: Have authors followed on the bleeding complications like epistaxis, menorrhagia, gum bleedings, etc.?			"It is still not clear how thrombophilia was defined. You write (page 3, lines 129-130): ""Only women with thrombophilia defined as a factor correlated with thrombotic phenomena linked to congenital or acquired factors. [21,40-43]"" I suggest you alter the sentence, e.g. to something like: ""Thrombophilia was defined as either the diagnosis of an inherited thrombophilia, a personal history of VTE or a familial history of VTE."" If that is how you defined thrombophilia?"	3	2	healthcare10030476_makarova	0
+To the Editor in Chief of Healthcare We re-submitted our article “The impact of direct oral anticoagulant prophylaxis for thromboembolism in thrombofilic patients underwent abdominoplastic surgery”, Manuscript ID: healthcare-1600106, Section: Environmental Factors and Global Health, Special issue: Skin Disorders in Hematological Disease.			Also, another round of language editing may be suitable.	3	2	healthcare10030476_makarova	0
+Also, “20210 gene” should be “20210 polymorphisms, or “G20210A polymorphism” to be completely correct.			"I have one major comment. It is still not clear how thrombophilia was defined. You write (page 3, lines 129-130): ""Only women with thrombophilia defined as a factor correlated with thrombotic phenomena linked to congenital or acquired factors. [21,40-43]"" I suggest you alter the sentence, e.g. to something like: ""Thrombophilia was defined as either the diagnosis of an inherited thrombophilia, a personal history of VTE or a familial history of VTE."" If that is how you defined thrombophilia?"	3	2	healthcare10030476_perova	0
+Point 5: The Discussion section could benefit from a language revision, especially in the latter part, to Answer to the Reviewer point 5: The observation of the reviewer has been accepted and the manuscript was modified accordingly.			Also, another round of language editing may be suitable.	3	2	healthcare10030476_perova	0
+C) Page 3, lines 133-134: “Patients were then divided into two groups according to the drugs employed in antithrombotic chemoprophylaxis:” How was it decided which patients received which form of prophylaxis?			In the Abstract you state that “two patients during their course of Rivaroxaban presented severe hematoma”, but in Table 1 it appears that the two hematomas occurred in the enoxaparin group? Please clarify.	1	2	healthcare10030476_perova	0
+2-3% of what for the 20210 polymorphism and 3-8% of what for FV Leiden?			The introduction is not well written. The structure is unclear, and the authors repeat themselves many times. For example, the section begins with introducing inherited thrombophilias and VTE risk in these conditions, then proceeds to surgery, then back again to thrombophilia where it repeats itself and even states other numbers for prevalences and VTE risks than previously stated. Furthermore, suitable references are lacking in some places. For the present study to be acceptable for publication, the introduction should be thoroughly revised.	1	2	healthcare10030476_perova	0
+Answer to the Reviewer point 8: The observation of the reviewer has been accepted and the manuscript was modified accordingly.			The Discussion section could benefit from a language revision, especially in the latter part, to make the flow better. Also, a brief discussion on the limitations of your study should be included.	1	2	healthcare10030476_perova	0
+In the abstract, lines 23-27, I suggest to use the following expression or something like that.			The citations are not formatted according to the journal guidelines. Please revise.	1	2	healthcare10040723_perova	0
+Also, the research questions lack describing the study setting.			Include the full form of STD described in the results.	1	2	healthcare10040723_perova	0
+Importantly, the study has pointed out the lack of in-depth awareness about digital health technologies in students and highlighted the need for education of the same.			Table 3 “Health apps and computer algorithms are for patients disturbing (=)…” should “(=)” be “(0)”?	1	2	healthcare10040723_perova	0
+However, the first sentence did not appear to serve this purpose.			Line 252, 24/7 is the commonly accepted form of 7/24.	1	2	healthcare10040723_perova	0
+For example, the first sentence should describe the general aim or the overall objective of the study in a more general term.			Line 267 and 268, the description of figure 1 is not clear. Does “focused on senses” mean in-person patient-doctor interaction?	1	2	healthcare10040723_perova	0
+Author Response See the point-by-point response in the attached Word file.			Table 4 data suggest Digital networks increase doctor nurse communication but Line 266 says it undermines the same. Can you clarify?	1	2	healthcare10040723_perova	0
+Phrases like ‘2020 medical students’ generation,’ and ‘their level on confidence,’ are confusing for the readers.			Table 5, what does numbers 1 through 7 represent?	1	2	healthcare10040723_perova	0
+Page 3, lines 117-118, the statement has two issues.			The fundamental issue that needs clarity is whether or not the study design was ‘exploratory mixed methods design’ or ‘convergent parallel design.’ As far as exploratory mixed methods design is concerned, the researcher begins with qualitative data and then collects quantitative information. Typically, in this design, the researcher presents the study in two phases, with the first phase involving qualitative data collection and the second phase involving quantitative data. In this regard, your study fits with the exploratory nature. In an exploratory sequential mixed-methods design, the researcher emphasizes the qualitative data (QUAL) more than the quantitative data (quan). This emphasis may reveal by presenting the overarching question as an open-ended question or discussing the qualitative results in more detail than the quantitative results. However, in this manuscript, I have not seen elements of these characteristics in the paper. As I have seen a complementary role of the qualitative and quantitative pieces is apparent instead of the qualitative piece informing the quantitative pieces. For example, the description of the study design says the following. Page 2, lines 90-92, ‘This was then integrated into the second study phase consisting of a nationally representative sample of the same sort of cohort.’ This description shows the integration of the two phases instead of the preceding phase informing the next phase. From the explanation in the study design and the presentation of your results, it sounds to me as if the mixed-methods nature of this study fits more with the convergent parallel design. Please, give your readers more information that conveniences the exploratory nature.	1	2	healthcare10040723_perova	0
+Importantly, the study has pointed out the lack of in-depth awareness about digital health technologies in students and highlighted the need for education of the same.			In the abstract, lines 23-27, I suggest to use the following expression or something like that. Around 38% of the students felt ill-prepared and could not answer AI-related questions because digitization in medicine and AI are not a formal part of the medical curriculum.	1	2	healthcare10040723_perova	0
+I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.			Page 2, line 42, ‘the computation of compounding factors,’ I did not understand what it means. I suggest the authors to replace this with appropriate phrase.	1	2	healthcare10040723_perova	0
+Importantly, the study has pointed out the lack of in-depth awareness about digital health technologies in students and highlighted the need for education of the same.			On page 2, line 111, the phrase ‘to affirm or dismiss,’ does not make sense. Replace this phrase with a more appropriate phrase.	1	2	healthcare10040723_perova	0
+Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement.			Page 3, lines 117-118, the statement has two issues. ‘The items were then tested through a pilot study consisting of a 117 group of 4 pre- and clinical students, mediated by AG and JE’ First, why did you use only 4 students for pilot testing? Second, what do ‘AG’ and ‘JE’ represent?	1	2	healthcare10040723_perova	0
+For example, the first sentence should describe the general aim or the overall objective of the study in a more general term.			Page 3, line 132, it says, Convenience sampling was used. I would ask, why did you use cconvenience sampling?	1	2	healthcare10040723_perova	0
+‘The items were then tested through a pilot study consisting of a 117 group of 4 pre- and clinical students, mediated by AG and JE’.			Page 4, table 1, the caption and the table content do not match. Revise either of them.	1	2	healthcare10040723_perova	0
+In the methods section, clearly describe the sampling procedure for the quantitative and qualitative study parts separately.			In the methods section, clearly describe the sampling procedure for the quantitative and qualitative study parts separately. Also describe the final study sample for both. I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.	1	2	healthcare10040723_perova	0
+1 Review comments Title: Medical students' perceptions towards digitization and artificial intelligence: A mixed methods study Thank you for the opportunity to review this manuscript.			Page 12, line 389, the statement which says, ‘The non-probabilistic sampling makes a generalization of the answers difficult,’ is confusing. I suggest to re-write this sentence.	1	2	healthcare10040723_perova	0
+I suggest to re-write the aims and study questions more clearly.			I suggest to remove the phrase ‘so called,’ that has been used in the conclusion section or any other section.	1	2	healthcare10040723_perova	0
+I suggest the authors to replace this with appropriate phrase.			Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement. Re-write this sentence clearly.	1	2	healthcare10040723_perova	0
+I hope these suggestions are of use to the authors.			I suggest the authors to include a short section including the implications of the study after the conclusions.	1	2	healthcare10040723_perova	0
+Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			In line 227 “back of tobacco” is not clear be more specific.	1	2	horticulturae8050412_makarova	0
+This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			Minor corrections In line 38 replace “Fortunately” with However.	1	2	horticulturae8050412_makarova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			In line 63 replace “trancroptoma” with transcriptome.	1	2	horticulturae8050412_makarova	0
+Point 4: Line 267-268: Please, check this sentence.			In line 65 replace “And” with In addition.	1	2	horticulturae8050412_makarova	0
+Response 3: Special thanks to you for your good suggestion.			In line 69 add period after assay.	1	2	horticulturae8050412_makarova	0
+In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China).			In line 83 what extraction method was used? Mention kit of reagent brand.	1	2	horticulturae8050412_makarova	0
+Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			In line 91 what brand was Taq polymerase from?	1	2	horticulturae8050412_makarova	0
+Point 17: In line 131 delete the period after experiments.			In line 100 replace “ligated” with was ligated and quote a reference for pORERA-GFP vector or mention brand if it is commercial.	1	2	horticulturae8050412_makarova	0
+"Point 7: In line 65 replace ""And"" with In addition."			In line 113 replace “psoup” with pSOUP and “tobaccos” with tobacco plants.	1	2	horticulturae8050412_makarova	0
+The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves.			In line 117 and 267 replace “vacuuming penetration” with vacuum infiltration.	1	2	horticulturae8050412_makarova	0
+In this view, I've only some few questions for the authors.			In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	1	2	horticulturae8050412_makarova	0
+"Point 6: In line 63 replace ""trancroptoma"" with transcriptome."			In line 129 replace “strains” and “strain” with lines and line.	1	2	horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			In line 131 delete the period after experiments.	1	2	horticulturae8050412_makarova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			In line 138 Seedling should be lowercase.	1	2	horticulturae8050412_makarova	0
+This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			In line 149 mention the kit brand.	1	2	horticulturae8050412_makarova	0
+In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China).			In line 154 add “strain” after receptor and mention kit brand.	1	2	horticulturae8050412_makarova	0
+"Point 16: In line 129 replace ""strains and ""strain"" with lines and line."			In line 169 add “were done” at the end. A phrase must have a verb.	1	2	horticulturae8050412_makarova	0
+In this view, I've only some few questions for the authors.			In line 261 replace “tobaccos” with tobacco plants.	1	2	horticulturae8050412_makarova	0
+"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			In line 262 replace “were treatment by” with were treated with.	1	2	horticulturae8050412_makarova	0
+Point 4: Line 267-268: Please, check this sentence.			In line 264 replace “indicate” with indicated.	1	2	horticulturae8050412_makarova	0
+Point 10: Figure 9 caption: measure instead of measuring.			In line 292 “The” must be lowercase.	1	2	horticulturae8050412_makarova	0
+Point 18: In line 138 Seedling should be lowercase.			In line 318 “We” must be lowercase.	1	2	horticulturae8050412_makarova	0
+"Point 13: In line 117 and 267 replace ""vacuuming penetration"" with vacuum infiltration."			In line 395 replace “it revealed” with revealed.	1	2	horticulturae8050412_makarova	0
+"Point 11: In line 100 replace ""ligated"" with was ligated and quote a reference for pORER4-GFP vector or mention brand if it is commercial."			In line 406 delete period after stress.	1	2	horticulturae8050412_makarova	0
+"Point 14: In line 118 define ""symbiotic medium""."			Line 302: plant leaves instead of plants leaves.	1	2	horticulturae8050412_makarova	0
+"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?	1	2	horticulturae8050412_makarova	0
+We have revised the manuscript according to your comments (Line 274-277).			Auhtors report that Actin was used as housekeeping gene. Do the authors have performed an efficiency test before using it? Do they tried to use otherHK genes?	1	2	horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			Furthermore, I reccomend a text revision since some errors are present in the main text. Line 124: generate instead of generate	1	2	horticulturae8050412_makarova	0
+"Point 25: In line 292 ""The"" must be lowercase."			"Line 267-268: Please, check this sentence. It ends with ""then"". Figure 5 caption: activates instead of activate Line 302: plant leaves instead of plants leaves"	1	2	horticulturae8050412_makarova	0
+Point 17: In line 131 delete the period after experiments.			Figure 5 caption: activates instead of activate.	1	2	horticulturae8050412_makarova	0
+"Point 23: In line 262 replace ""were treatment by"" with were treated with."			Line 302: plant leaves instead of plants leaves.	1	2	horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			Line 311: plant leaves instead of plants leaves	1	2	horticulturae8050412_makarova	0
+"Point 26: In line 318 ""We"" must be lowercase."			"Line 315: maybe a semicolon after ""salt stress"" is appropriate."	1	2	horticulturae8050412_makarova	0
+"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			"Line 360: remove the space after ""assay"""	1	2	horticulturae8050412_makarova	0
+"Point 25: In line 292 ""The"" must be lowercase."			Figure 9 caption: measure instead of measuring	1	2	horticulturae8050412_makarova	0
+In this view, I've only some few questions for the authors.			"Line 392: I suggest to change the sentence in the following way "".. to drive the expression of target"	1	2	horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			"Line 446: ""...participate in salt stress""Line 448: check the word ""redycing"" "	1	2	horticulturae8050412_makarova	0
+Point 17: In line 131 delete the period after experiments.			In line 227 “back of tobacco” is not clear be more specific.	1	2	horticulturae8050412_perova	0
+2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.			Rephrase lines 317-318. Comments as above.	1	2	horticulturae8050412_perova	0
+Results clearly describe the protein localization as well as its role in stress response.			In lines 271-272 and 395-396, how do you explain that salt induced promoter trans-activation? Is there any published precedent?	1	2	horticulturae8050412_perova	0
+; Bustin, S.A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR.			Write appropriate brand of kits and reagents, followed by country. Check journal format.	1	2	horticulturae8050412_perova	0
+2005, 139: 5-17. https://doi.org/10.1104/pp.105.063743 Hou, H.; Jia, H.; Yan, Q.; Wang, X. Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance.			In line 38 replace “Fortunately” with However.	1	2	horticulturae8050412_perova	0
+Point 17: In line 131 delete the period after experiments.			In line 63 replace “trancroptoma” with transcriptome.	1	2	horticulturae8050412_perova	0
+Results clearly describe the protein localization as well as its role in stress response.			In line 65 replace “And” with In addition.	1	2	horticulturae8050412_perova	0
+In Silico and Transcription Analysis of Trehalose-6-phosphate Phosphatase Gene Family of Wheat: Trehalose Synthesis Genes Contribute to Salinity, Drought Stress and Leaf Senescence.			In line 69 add period after assay.	1	2	horticulturae8050412_perova	0
+Point 9: In line 83 what extraction method was used?			In line 83 what extraction method was used? Mention kit of reagent brand.	1	2	horticulturae8050412_perova	0
+Response 7: Special thanks to you for your good suggestion.			In line 91 what brand was Taq polymerase from?	1	2	horticulturae8050412_perova	0
+"Point 23: In line 262 replace ""were treatment by"" with were treated with."			In line 100 replace “ligated” with was ligated and quote a reference for pORERA-GFP vector or mention brand if it is commercial.	1	2	horticulturae8050412_perova	0
+Point 10: Figure 9 caption: measure instead of measuring.			In line 113 replace “psoup” with pSOUP and “tobaccos” with tobacco plants.	1	2	horticulturae8050412_perova	0
+2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X.			In line 117 and 267 replace “vacuuming penetration” with vacuum infiltration.	1	2	horticulturae8050412_perova	0
+Furthermore, I reccomend a text revision since some errors are present in the main text.			In line 118 define “symbiotic medium”.	1	2	horticulturae8050412_perova	0
+Point 6: Line 302: plant leaves instead of plants leaves.			In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	1	2	horticulturae8050412_perova	0
+Point 18: In line 138 Seedling should be lowercase.			In line 129 replace “strains” and “strain” with lines and line.	1	2	horticulturae8050412_perova	0
+"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			In line 131 delete the period after experiments.	1	2	horticulturae8050412_perova	0
+"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			In line 138 Seedling should be lowercase.	1	2	horticulturae8050412_perova	0
+Point 9: In line 83 what extraction method was used?			In line 149 mention the kit brand.	1	2	horticulturae8050412_perova	0
+"Point 27: In line 395 replace ""it revealed"" with revealed."			In line 154 add “strain” after receptor and mention kit brand.	1	2	horticulturae8050412_perova	0
+"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			In line 169 add “were done” at the end. A phrase must have a verb.	1	2	horticulturae8050412_perova	0
+This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			In line 261 replace “tobaccos” with tobacco plants.	1	2	horticulturae8050412_perova	0
+In this view, I've only some few questions for the authors.			In line 262 replace “were treatment by” with were treated with.	1	2	horticulturae8050412_perova	0
+"Point 23: In line 262 replace ""were treatment by"" with were treated with."			In line 264 replace “indicate” with indicated.	1	2	horticulturae8050412_perova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			In line 292 “The” must be lowercase.	1	2	horticulturae8050412_perova	0
+Response 10: Special thanks to you for your good suggestion.			In line 318 “We” must be lowercase.	1	2	horticulturae8050412_perova	0
+Furthermore, I reccomend a text revision since some errors are present in the main text.			In line 395 replace “it revealed” with revealed.	1	2	horticulturae8050412_perova	0
+Results clearly describe the protein localization as well as its role in stress response.			Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?	1	2	horticulturae8050412_perova	0
+"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			Auhtors report that Actin was used as housekeeping gene. Do the authors have performed an efficiency test before using it? Do they tried to use otherHK genes?	1	2	horticulturae8050412_perova	0
+A deeeping on protein an nucleotide structures was also reported.			Line 124: generate instead of generate	1	2	horticulturae8050412_perova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			"Line 267-268: Please, check this sentence. It ends with ""then"".
+Reviewer_2	Author	1	5	0	2	Figure 5 caption: activates instead of activate
+Reviewer_2	Author	1	6	0	2	Line 302: plant leaves instead of plants leaves
+Reviewer_2	Author	1	7	0	2	Line 311: plant leaves instead of plants leaves
+Reviewer_2	Author	1	8	0	2	Line 315: maybe a semicolon after """"salt stress"""" is appropriate"""	1	2	horticulturae8050412_perova	0
+This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			"Line 360: remove the space after ""assay"""	1	2	horticulturae8050412_perova	0
+Point 7: Line 311: plant leaves instead of plants leaves.			Figure 9 caption: measure instead of measuring	1	2	horticulturae8050412_perova	0
+Point 18: In line 138 Seedling should be lowercase.			"Line 392: I suggest to change the sentence in the following way "".. to drive the expression of target genes.."""	1	2	horticulturae8050412_perova	0
+2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.			"Line 446: ""...participate in salt stress"""	1	2	horticulturae8050412_perova	0
+Response 4: We feel sorry for the unclear statement.			"Line 448: check the word ""redycing"""	1	2	horticulturae8050412_perova	0
+In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way.			Lines 116-118: The amounts of samples were confused. Although there were 66 children involved in this study, 66 food samples, 62 feces and 64 urine samples were collected.	1	2	ijerph19127332_makarova	0
+Lines 116-118: The amounts of samples were confused.			Lines 176-178: Different units between manuscript and SM (figure S1).	1	2	ijerph19127332_makarova	0
+At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd.			Lines 209-211: Different SIR values between manuscript and figure 1.	1	2	ijerph19127332_makarova	0
+Heavy metal elements accumulate in the 36 human body and interfere with the human endocrine system [5], damage the body's car- 37 diovascular and nervous systems [6.7], and can even lead to cancer [8].			Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.	1	2	ijerph19127332_makarova	0
+In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way.			Lines 218 and 223: check the Figure 3 and figure S1.	1	2	ijerph19127332_makarova	0
+The authors should add section(s) for the heavy metal concentrations in biotic samples.			Lines 253 and 377: different values between SIR and figure 3.	1	2	ijerph19127332_makarova	0
+The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3.			Lines 348-349: correct the order of HQs.	1	2	ijerph19127332_makarova	0
+Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.			correct the figure caption in SM.	1	2	ijerph19127332_makarova	0
+Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas.			Title- ‘in’ should be used for larger areas, can you add the study locations? Otherwise replace ‘in’ with ‘at’Rephrase title so as not to repeat ‘and’ Answer:	1	2	ijerph19127332_makarova	0
+S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”.			It was only mentioned towards the end – Discussion Answer: Thanks for your suggestion. Materials and Methods:I would like to see a better description of the study location.	1	2	ijerph19127332_makarova	0
+Answer: Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L.			Better explanation needed - why different processing for different samples?	1	2	ijerph19127332_makarova	0
+please rephrase Answer: Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities” Point 11.			Line190 – why was 350d/y taken? Explain for better understanding	1	2	ijerph19127332_makarova	0
+Finally, the digest was analyzed by HR‒ICP‒MS.			Will like to see ethical approval for the project/research? Ethical approval from which institution? Were Consent/assent from participants obtained?	1	2	ijerph19127332_makarova	0
+Also describe in the methods section Answer: Thank you for your proposal.			Results and Discussion: Table 1 – is it possible to list/ summarise the data for each day?	1	2	ijerph19127332_makarova	0
+Finally, the digest was analyzed by HR‒ICP‒MS.			Line 294 – what is meaning of BTM, Would prefer re-introduction of the full meaning of acronyms in each sub-section Answer: Thank you for your suggestions.	1	2	ijerph19127332_makarova	0
+Health risk assessments showed that Cr 23 presented the greatest carcinogenic risk more than 10‒6 in this typical polluted area, while As was 24 second.			Line 320 -Also descript the study location in the method section Answer: Thank you for your suggestions.	1	2	ijerph19127332_makarova	0
+Table 1 – is it possible to list/ summarise the data for each day?			Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.	1	2	ijerph19127332_makarova	0
+Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills.			Line 346 – What is HQ? Also describe in the methods section Answer:	1	2	ijerph19127332_makarova	0
+Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min.			Line 368 – what is the meaning of ‘Whatever’? please rephrase Answer:	1	2	ijerph19127332_makarova	0
+Lines 209-211: Different SIR values between manuscript and figure 1.			Will like to see references on the metal concentrations at the e-waste recycling site.	1	2	ijerph19127332_makarova	0
+This study estimated the heavy metal exposure to children through soil and dust Ingestion in electronic waste (e‒waste) dismantling areas.			Page 2, lines 76-78: You can remove this sentence: a class of elements in the human body that are not easily absorbed by the human gastrointestinal tract and are also difficult to be transformed into other substances it’s not necessary Answer:	1	2	ijerph19127332_makarova	0
+This method has been used before”, “the Hazard Quotients (HQ) values between 1 and 10 indicate likely damage to human health”.			Page 3, lines 99-100: please, add references for the following information South China's economic conditions are better than in North China, but the environmental pollution is worse. The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.	1	2	ijerph19127332_makarova	0
+Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Author Response File: Author Response.docx			Freeze-drying conditions of feces. Page 3, lines 116 and 120: Please, add technical details about: lyophilization of food samples.	1	2	ijerph19127332_makarova	0
+Page 3, lines 121-124:Please, add some information: How did you collect dust samples?			How much soil (kg) did you collect for each sample? Was the soil quartered? Page 3, lines 121-124: Please, add some information: How did you collect dust samples?	1	2	ijerph19127332_makarova	0
+The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.			Lines 137-138: Please, modify as suggested The supernatant was then extracted and analyzed by HR‒ICP‒MS to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y, Answer: Thanks for your advice!	1	2	ijerph19127332_makarova	0
+Other outliers may be due to the behavior of certain children, such as pica behavior or spending unusually large amounts of time playing in grassland.			Please, add labels (a, b, c, etc.) in each figure and update the captions. Also in the supplementary. Please, check captions of figures S2 and 3 that are identical.	1	2	ijerph19127332_makarova	0
+Please, use element symbols not the entire name in all the manuscript.			Lines 116-118: The amounts of samples were confused. Although there were 66 children involved in this study, 66 food samples, 62 feces and 64 urine samples were collected.	1	2	ijerph19127332_perova	0
+Thus, we cannot provide the data for each day.			Lines 176-178: Different units between manuscript and SM (figure S1).	1	2	ijerph19127332_perova	0
+The results provided insights into the varying health risks of different have metals.			Lines 209-211: Different SIR values between manuscript and figure 1.	1	2	ijerph19127332_perova	0
+Health risk assessments showed that Cr 23 presented the greatest carcinogenic risk more than 10‒6 in this typical polluted area, while As was 24 second.			Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.	1	2	ijerph19127332_perova	0
+The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.			Lines 218 and 223: check the Figure 3 and figure S1.	1	2	ijerph19127332_perova	0
+Line 368 – what is the meaning of ‘Whatever’?			Lines 253 and 377: different values between SIR and figure 3.	1	2	ijerph19127332_perova	0
+S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”.			Lines 348-349: correct the order of HQs.	1	2	ijerph19127332_perova	0
+The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y.			correct the figure caption in SM.	1	2	ijerph19127332_perova	0
+The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas.			Title- ‘in’ should be used for larger areas, can you add the study locations? Otherwise replace ‘in’ with ‘at’Rephrase title so as not to repeat ‘and’ Answer:	1	2	ijerph19127332_perova	0
+Finally, the digest was analyzed by HR‒ICP‒MS.			I would like to see a better description of the study location. It was only mentioned towards the end – Discussion Answer: Thanks for your suggestion.	1	2	ijerph19127332_perova	0
+Temperature of the heating plate for faces sample digestion.			Better explanation needed - why different processing for different samples?	1	2	ijerph19127332_perova	0
+In our study, 66 children were taken part in our analysis.			Line190 – why was 350d/y taken? Explain for better understanding Answer:	1	2	ijerph19127332_perova	0
+Page 3, lines 131-151: Please, add some information:How many millilitres of HNO3, H2O2, HF, HClO4 did you use for the digestion procedures (food samples, faces samples, soil and dust samples)?			Table 1 – is it possible to list/ summarise the data for each day?	1	2	ijerph19127332_perova	0
+Lines 176-178: Different units between manuscript and SM (figure S1).			Line 294 – what is meaning of BTM, Would prefer re-introduction of the full meaning of acronyms in each sub-section Answer: Thank you for your suggestions.	1	2	ijerph19127332_perova	0
+Page 3, lines 121-124: Please, add some information: How did you collect dust samples?			Line 320 -Also descript the study location in the method section Answer: Thank you for your suggestions.	1	2	ijerph19127332_perova	0
+Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.			Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.	1	2	ijerph19127332_perova	0
+Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills.			Line 346 – What is HQ? Also describe in the methods section Answer:	1	2	ijerph19127332_perova	0
+International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428 Point 3.			Line 368 – what is the meaning of ‘Whatever’? please rephrase Answer:	1	2	ijerph19127332_perova	0
+Better explanation neededLine190 – why was 350d/y taken?			Will like to see references on the metal concentrations at the e-waste recycling site.	1	2	ijerph19127332_perova	0
+The digestion process is repeated until the sample becomes sticky.			Page 2, lines 76-78: You can remove this sentence: a class of elements in the human body that are not easily absorbed by the human gastrointestinal tract and are also difficult to be transformed into other substances it’s not necessary Answer:	1	2	ijerph19127332_perova	0
+Lines 253 and 377: different values between SIR and figure 3.			Page 3, lines 99-100: please, add references for the following information South China's economic conditions are better than in North China, but the environmental pollution is worse. The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.	1	2	ijerph19127332_perova	0
+I suggested the major revision before publication.			Page 3, lines 116 and 120: Please, add technical details about:lyophilization of food samples. Freeze-drying conditions of feces.	1	2	ijerph19127332_perova	0
+The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 6.			Page 3, lines 121-124:Please, add some information: How did you collect dust samples? How much soil (kg) did you collect for each sample? Was the soil quartered?	1	2	ijerph19127332_perova	0
+Line 320 -Also descript the study location in the method section Answer: Thank you for your suggestions.			Lines 131-135: Please, modify as suggested Crushed food samples (1 g) were microwaved with concentrated nitric acid and hydrogen peroxide (HNO3‒H2O2). The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Lines 137-138: Please, modify as suggested The supernatant was then extracted and analyzed by HR‒ICP‒MS to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y, Answer: Thanks for your advice!	1	2	ijerph19127332_perova	0
+However, we used the data without outlier to construct the figure.			Please, add labels (a, b, c, etc.) in each figure and update the captions. Also in the supplementary. Please, check captions of figures S2 and 3 that are identical.	1	2	ijerph19127332_perova	0
+Thank you for improving this paper, especially regarding the timing of TOC administration. Still,			The impact of Tocilizumab on radiological changes can not be adequately analyzed without a control group not receiving the drug.	1	2	jcm11051247_makarova	0
+The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).			The proposed timing of Tocilizumab administration based on oxygen flow rate is interesting but undoubtedly requires a more detailed statistical analysis to convince the readers.	1	2	jcm11051247_makarova	0
+Comparative radiologic changes before and after TOC administration In Table 2, about the change in CT findings in fatal and non-fatal patients before and after TOC administration, isn't it the same as the difference in imaging findings between patients who survive after severe illness and those who do not, regardless of TOC?			Materials and Methods 2.1. Study design and population If there are mutations or differences in viral strains during the course of the study, should differences in response to TOC and timing of administration depending on the strain be considered?	1	2	jcm11051247_makarova	0
+When comparing different oxygen delivery methods, isn't it more appropriate to compare FiO2 rather than oxygen flow rate?			Results 86 cases are excluded from 187 cases, which is too many. What is the reason?	1	2	jcm11051247_makarova	0
+Comparative radiologic changes before and after TOC administration In Table 2, about the change in CT findings in fatal and non-fatal patients before and after TOC administration, isn't it the same as the difference in imaging findings between patients who survive after severe illness and those who do not, regardless of TOC?			Results When comparing different oxygen delivery methods, isn't it more appropriate to compare FiO2 rather than oxygen flow rate?	1	2	jcm11051247_makarova	0
+Point 2: The proposed timing of Tocilizumab administration based on oxygen flow rate is interesting but undoubtedly requires a more detailed statistical analysis to convince the readers.			Results The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).	1	2	jcm11051247_makarova	0
+Study design and population If there are mutations or differences in viral strains during the course of the study, should differences in response to TOC and timing of administration depending on the strain be considered?			The oxygen flow rate of 13 L/min measured by ROC as a cutoff value for TOC administration should be presented by FiO2.	1	2	jcm11051247_makarova	0
+The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).			Comparative radiologic changes before and after TOC administration In Table 2, about the change in CT findings in fatal and non-fatal patients before and after TOC administration, isn't it the same as the difference in imaging findings between patients who survive after severe illness and those who do not, regardless of TOC?	1	2	jcm11051247_makarova	0
+Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.			Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.	1	2	jcm11051247_makarova	0
+Should the writing of the results and discussion sections be separated?			Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg. prediction mechanism, prediction accuracy, etc.) is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.	1	2	land11030363_makarova	0
+Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.			When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio. There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?	1	2	land11030363_makarova	0
+Line 264: “…, and hence parent material.” It’s not clear here.			Should the writing of the results and discussion sections be separated? Please think carefully.	1	2	land11030363_makarova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).	1	2	land11030363_makarova	0
+The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			Line 104, 195, 223, 544: Is it “modelling” or “modeling”? The whole text should be unified.	1	2	land11030363_makarova	0
+Should the writing of the results and discussion sections be separated?			Lines 145-146: “, soil reaction (pH) in 1:1 soil to KCl solution” is not clearly expressed.	1	2	land11030363_makarova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence? Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources. What does the controlled laboratory environment you describe look like?	1	2	land11030363_makarova	0
+"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			Line 174: “…resulting in 12 spectra”. It is not clear how the 12 spectra were obtained here.	1	2	land11030363_makarova	0
+In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are.			Line 176: “…resulted 50 spectra…”. It is also not clear how 50 spectra were obtained.	1	2	land11030363_makarova	0
+prediction mechanism, prediction accuracy, etc.)			Line 190: “…moving window sizes (mainly 7, 9 and 11)”. Need to use 3 different moving window sizes? It undoubtedly increases your experimental workload, and you do not explain it later. As far as I know, most articles use only one moving window size.	1	2	land11030363_makarova	0
+My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”. Do these two places mean the same thing?	1	2	land11030363_makarova	0
+It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1.			Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.	1	2	land11030363_makarova	0
+This may help to analyze the underlying mechanism.			Line 264: “…, and hence parent material.” It’s not clear here.	1	2	land11030363_makarova	0
+Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?			Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?	1	2	land11030363_makarova	0
+Too many parentheses are used in the essay, which is not recommended in writing.			Lines 325-328: “Regardless of… spectra.” Is there a necessary connection between these two sentences? The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.	1	2	land11030363_makarova	0
+Line 347: “different pattern” should be “different patterns”.			Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.	1	2	land11030363_makarova	0
+However, there are some issues that need to be resolved before this manuscript can be considered for publication.			Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here. And all in Table 3 are “10-bands”.	1	2	land11030363_makarova	0
+Reference to formal analytical methods must be made in a scientific manner.			Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	1	2	land11030363_makarova	0
+In addition, it needs support with more and more up-to-date recent articles.			Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	1	2	land11030363_makarova	0
+Line 135: “impairment water resources” Is this expression appropriate?			Line 449, 479: “CO3”?	1	2	land11030363_makarova	0
+Please pay attention to the singular and plural in the sentence.			Line 455: “soil mineralogy”?	1	2	land11030363_makarova	0
+"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			Lines 456-457: What does this sentence do here?	1	2	land11030363_makarova	0
+This may help to analyze the underlying mechanism.			Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.	1	2	land11030363_makarova	0
+Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what? Readers may be more interested in your analysis and interpretation.	1	2	land11030363_makarova	0
+Lines 456-457: What does this sentence do here?			Line 520: The correlation between K content and P content is not very high (r = 0.40*), but their corresponding VIP patterns are “identical”, how do you explain it?	1	2	land11030363_makarova	0
+My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			Line 523: “correspond absorption” should be “correspond to absorption”.	1	2	land11030363_makarova	0
+Table 1: The pH values are different from those in the previous version.			Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence? Is VIP peak related to Fe or SOC content?	1	2	land11030363_makarova	0
+c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned.			Many thanks for your response. The manuscript has been largely improved. Modified content should be marked in the revised manuscript for easy review by reviewers.	3	2	land11030363_makarova	0
+Table 1: The pH values are different from those in the previous version.			The abstract could be more concise.	3	2	land11030363_makarova	0
+Line 135: “impairment water resources” Is this expression appropriate?			The introduction part can highlight the key points and innovations of this paper.	3	2	land11030363_makarova	0
+Too many parentheses are used in the essay, which is not recommended in writing.			The definite article “the” is missing in many places in the whole article. please check it.	3	2	land11030363_makarova	0
+As far as I know, most articles use only one moving window size.			Too many parentheses are used in the essay, which is not recommended in writing.	3	2	land11030363_makarova	0
+It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1.			Line 135: “impairment water resources” Is this expression appropriate?	3	2	land11030363_makarova	0
+is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			Table 1: The pH values are different from those in the previous version.	3	2	land11030363_makarova	0
+Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.			Attention should be paid to the number of decimal places in all numbers throughout the text.	3	2	land11030363_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit. Please check the format of all references.	3	2	land11030363_makarova	0
+Attention should be paid to the number of decimal places in all numbers throughout the text.			The conclusion is too long and does not highlight the key results.	3	2	land11030363_makarova	0
+However, there are some issues that need to be resolved before this manuscript can be considered for publication.			The novelty of the study should be clearly stated in the Introduction section after the objectives are formulated.	1	2	land11030363_makarova	0
+Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses. Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments."	1	2	land11030363_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			b) It is necessary to explain the method of sampling using the appropriate literature. Is the number of samples sufficient for the purpose of the research? Please explain.	1	2	land11030363_makarova	0
+In addition, it needs support with more and more up-to-date recent articles.			c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned. Reference to formal analytical methods must be made in a scientific manner. The writing language used must be different from the way an analyst mentions it in the lab.	1	2	land11030363_makarova	0
+Please pay attention to the singular and plural in the sentence.			The “Introduction section ” should be reworked, highlighting the relevance of this topic worldwide. It would be wise to give more international flavor in the sections of Introduction and Discussion.	1	2	land11030363_makarova	0
+Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			In addition, it needs support with more and more up-to-date recent articles. I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research,https://doi.org/10.1007/s11356-021-14673-0	1	2	land11030363_makarova	0
+Table 4 is related to partial correlation analysis with consideration of spkoil properties, and Figure 4 is related to the principal component analysis.			This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models. Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear. In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are. The abstract and conclusion parts lack conciseness and prominence. Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg. prediction mechanism, prediction accuracy, etc.) is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.	1	2	land11030363_perova	0
+Specific comments: When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio.			When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio. There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?	1	2	land11030363_perova	0
+Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit.			Should the writing of the results and discussion sections be separated? Please think carefully.	1	2	land11030363_perova	0
+Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.			During the discussion, the reasons behind the experimental results should be analyzed, not only from the correlation between the sample data but also cannot be explained by relying too much on the VIP score curve. In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide). This may help to analyze the underlying mechanism.	1	2	land11030363_perova	0
+The abstract and conclusion parts lack conciseness and prominence.			Line 104, 195, 223, 544: Is it “modelling” or “modeling”? The whole text should be unified.	1	2	land11030363_perova	0
+The abstract and conclusion parts lack conciseness and prominence.			Lines 145-146: “, soil reaction (pH) in 1:1 soil to KCl solution” is not clearly expressed.	1	2	land11030363_perova	0
+"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence? Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources. What does the controlled laboratory environment you describe look like?	1	2	land11030363_perova	0
+6) Discussion with new 3 subheading was significantly expanded and linked to the references, and the novelty was highlighted.			Line 174: “…resulting in 12 spectra”. It is not clear how the 12 spectra were obtained here.	1	2	land11030363_perova	0
+The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.			Line 176: “…resulted 50 spectra…”. It is also not clear how 50 spectra were obtained.	1	2	land11030363_perova	0
+In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are.			Line 190: “…moving window sizes (mainly 7, 9 and 11)”. Need to use 3 different moving window sizes? It undoubtedly increases your experimental workload, and you do not explain it later. As far as I know, most articles use only one moving window size.	1	2	land11030363_perova	0
+Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”. Do these two places mean the same thing?	1	2	land11030363_perova	0
+In addition, it needs support with more and more up-to-date recent articles.			Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.	1	2	land11030363_perova	0
+Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			Line 264: “…, and hence parent material.” It’s not clear here.	1	2	land11030363_perova	0
+Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?	1	2	land11030363_perova	0
+Modified content should be marked in the revised manuscript for easy review by reviewers.			Lines 325-328: “Regardless of… spectra.” Is there a necessary connection between these two sentences? The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.	1	2	land11030363_perova	0
+Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.	1	2	land11030363_perova	0
+Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg.			Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here. And all in Table 3 are “10-bands”.	1	2	land11030363_perova	0
+Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	1	2	land11030363_perova	0
+Modified content should be marked in the revised manuscript for easy review by reviewers.			Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	1	2	land11030363_perova	0
+My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			Line 449, 479: “CO3”?	1	2	land11030363_perova	0
+Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.			Line 455: “soil mineralogy”?	1	2	land11030363_perova	0
+There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?			Lines 456-457: What does this sentence do here?	1	2	land11030363_perova	0
+Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.	1	2	land11030363_perova	0
+During the discussion, the reasons behind the experimental results should be analyzed, not only from the correlation between the sample data but also cannot be explained by relying too much on the VIP score curve.			Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what? Readers may be more interested in your analysis and interpretation.	1	2	land11030363_perova	0
+Due to the separating the Results and Discussion section, overlapping of some comments made by reviewers and associated changes, all the text of the manuscript was modified.			Line 520: The correlation between K content and P content is not very high (r = 0.40*), but their corresponding VIP patterns are “identical”, how do you explain it?	1	2	land11030363_perova	0
+Line 135: “impairment water resources” Is this expression appropriate?			Line 523: “correspond absorption” should be “correspond to absorption”.	1	2	land11030363_perova	0
+However, there are some issues that need to be resolved before this manuscript can be considered for publication.			Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence? Is VIP peak related to Fe or SOC content?	1	2	land11030363_perova	0
+Line 264: “…, and hence parent material.” It’s not clear here.			Lines 541-543: Is this sentence appropriate here?	1	2	land11030363_perova	0
+"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			The abstract could be more concise.	3	2	land11030363_perova	0
+Line 264: ���…, and hence parent material.” It’s not clear here.			The introduction part can highlight the key points and innovations of this paper.	3	2	land11030363_perova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			The definite article “the” is missing in many places in the whole article. please check it.	3	2	land11030363_perova	0
+Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			Too many parentheses are used in the essay, which is not recommended in writing.	3	2	land11030363_perova	0
+Table 4 is related to partial correlation analysis with consideration of spkoil properties, and Figure 4 is related to the principal component analysis.			Line 135: “impairment water resources” Is this expression appropriate?	3	2	land11030363_perova	0
+This may help to analyze the underlying mechanism.			Table 1: The pH values are different from those in the previous version.	3	2	land11030363_perova	0
+6) Discussion with new 3 subheading was significantly expanded and linked to the references, and the novelty was highlighted.			Attention should be paid to the number of decimal places in all numbers throughout the text.	3	2	land11030363_perova	0
+prediction mechanism, prediction accuracy, etc.)			Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit. Please check the format of all references.	3	2	land11030363_perova	0
+Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			The conclusion is too long and does not highlight the key results.	3	2	land11030363_perova	0
+Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			The novelty of the study should be clearly stated in the Introduction section after the objectives are formulated.	1	2	land11030363_perova	0
+Line 264: “…CaCO3 and Fe),” Where is the left parenthesis?			There is not adequate information regarding quality control of the chemical analyses. Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.	1	2	land11030363_perova	0
+6) Discussion with new 3 subheading was significantly expanded and linked to the references, and the novelty was highlighted.			It is necessary to explain the method of sampling using the appropriate literature. Is the number of samples sufficient for the purpose of the research? Please explain.	1	2	land11030363_perova	0
+However, there are some important issues that need to be addressed:			Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned. Reference to formal analytical methods must be made in a scientific manner. The writing language used must be different from the way an analyst mentions it in the lab.	1	2	land11030363_perova	0
+Line 347: “different pattern” should be “different patterns”.			The “Introduction section ” should be reworked, highlighting the relevance of this topic worldwide. It would be wise to give more international flavor in the sections of Introduction and Discussion.	1	2	land11030363_perova	0
+In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are.			In addition, it needs support with more and more up-to-date recent articles. I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research,https://doi.org/10.1007/s11356-021-14673-0	1	2	land11030363_perova	0
+Author Response Reviewer 3 (R3): I thank the authors for submitting a revised version of their manuscript entitled “Self-reported long COVID in the general population: sociodemographic and health correlates in a cross-national sample”.			The authors admit that, line 367, younger people were overrepresented, but to me, it was that older persons, especially over age 70, who were somehow missed in the survey sampling process. The study offers little to say about long covid among the elderly.	1	2	life12060901_makarova	0
+**************************************************************************** Reviewer 3 (R3): The reviewed paper deals with the COVID pandemic, especially with the “long COVID” phenomenon.			The authors consider a number of factors associated with long COVID and psychological symptoms. I would suggest another factor - frustration with not getting well.	1	2	life12060901_makarova	0
+Psychological distress is the latent construct which is measured with the instrument GHQ.			I accept the manuscript for its publication in this journal.	1	2	life12060901_makarova	0
+Like being sick and tired of being sick and tired.			However, I see several issues with the ending of this paper. I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential. However, what I disliked was the interpretation and discussion of the empirical results. Frankly speaking, the authors are overselling their results. I found two major issues in the discussion of this paper: Authors response:	1	2	life12060901_makarova	0
+The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%).			The authors said in the statistic section that “statistical significance was set at p<0.05.” (page 5, line 212). However, later they wrote “The descriptively higher proportion of females with long COVID bordered statistical significance (p=0.05)” page 5, line 230) and “However, the different proportions of men and women with long COVID (22% and 33%, respectively) bordered towards statistical significance […]” (page 10, line 318). As a statistician, I felt very mad about this, because the results speak a different language, this is non-significant result.	1	2	life12060901_makarova	0
+Authors’ response: Thank you for pointing this out.			The authors wrote later in the conclusion section “[…] but a non-significant trend was found for the association between female gender and long COVID.” (page 11, line 389). This is a “no-go” in scientific research, the interpretation of “trends” resulted from non-significant results is strongly misleading. Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05). That is the true story. Hence, I urge the authors not to mislead readers and to re-write these sections.	1	2	life12060901_makarova	0
+I would suggest another factor - frustration with not getting well.			The next major issue targets the interpretation of interaction effects. The authors did three models with three different outcomes, namely psychological distress, fatigue, and perceived stress. In the results section, only two (and not three!) interaction effects became significant (psychological distress: p<0.001, fatigue: p<0.05, and perceived stress: p=0.36, see Table 4, page 7, line 269). However, the authors wrote “While participants with long COVID generally perceived more psychological distress, fatigue, and stress than those without long COVID, differences were larger for men than for women” page 9, line 303), Authors’ response: Authors introduced an interaction effect long COVID x gender into the model in order to test their third hypothesis (“(iii) whether gender moderated the associations between long 119 COVID status and the health outcomes”, page 3, line 119).	1	2	life12060901_makarova	0
+The reviewed paper deals with the COVID pandemic, especially with the “long COVID” phenomenon.			“[…] men’s perception of poorer health when having long COVID appears to include higher levels of psychological distress, fatigue and perceived stress” (page 11, line 363),	1	2	life12060901_makarova	0
+Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e.			and finally “Third, long COVID appears to have a stronger effect on men than on women” (page 11, line 401). Again, the authors are overselling their results. All these statements are not true, only for two out of three outcomes, namely psychological distress and fatigue, but not for perceived stress (see also Figure 3). Hence, I strongly urge the authors to re-write these sentences so that readers are not misled.	1	2	life12060901_makarova	0
+R3: “[…] and the negative effects of long COVID on the health outcomes were larger for men than for women” (page 11, line 392) Authors’ response: We have revised the sentence; see section 5.			Besides these two major issues, I detected some smaller errors and miss-spellings (note, this list in not complete). Hence, I have some comments/suggestions that I hope will help the authors to further develop this line of work: Authors’ response:	1	2	life12060901_makarova	0
+The authors did three models with three different outcomes, namely psychological distress, fatigue, and perceived stress.			Chapter 2 Materials and Methods: Something strange happened to the alignment of heading “2.2 Sample” (page 3, line 128).	1	2	life12060901_makarova	0
+Authors’ response: Thank you for pointing this out.			Please fix this. Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”). The total sample is reported here as 303. However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310). I know, there were 7 missing values within the 310. However, this table is still wrong, it must be 310 and not 303. The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%). Hence, I urge the authors to re-calculate percentages in Table 1. Otherwise, this is misleading the readership.	1	2	life12060901_makarova	0
+interaction effects became significant (psychological distress: p<0.001, fatigue: p<0.05, and perceived stress: p=0.36, see Table 4, page 7, line 269).			Chapter 2.3 Measures: Sometimes the verbal scales are set in italic (e.g., “0=better than ususal”, page 4, line 173) and sometimes not (e.g., “4=very often”, page 5, line 187).	1	2	life12060901_makarova	0
+We have corrected the manuscript according to this guidance.			Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold. For example, see Table 4 (page 7, line 269): Column Fatgigue, why is ES not printed in bold? Please use a consistent writing style.	1	2	life12060901_makarova	0
+ and finally “Third, long COVID appears to have a stronger effect on men than on women” (page 11, line 401).			Please use a consistent writing style throughout the whole manuscript. Results (page 7, line 274): Sometimes fatigue (page 7, line 274) and sometimes with a capital F (Fatigue, page 7, line 283).	1	2	life12060901_makarova	0
+I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential.			Wrong column headings in tables: I detected that the authors replaced “GHQ” with “psychological distress” in the text (also “PSS” replaced with “perceived stress”). This is correct. Psychological distress is the latent construct which is measured with the instrument GHQ. Unfortunately, the authors missed to replace GHQ by psychological distress in Table 4 (page 7, line 280) and Table 5 (page 7, line 297; see column headings: not “GHQ”, “Fatigue”, and “Perceived stress” but rather “Psychological distress”, “Fatigue”, and “Perceived stress”). Please correct the two tables.	3	2	life12060901_makarova	0
+Hence, I urge the authors not to mislead readers and to re-write these sections.			"Wrong use of abbreviations: The authors introduced the abbreviation “GHQ-12” for the 12-item General Health Questionnaire (see page 4, line 161), but wrote in the text only ""GHQ"" (e.g., page 5, line 207; page 6, line 256; page 7, line 274). Please replace every single “GHQ” in the text with the correct abbreviation “GHQ-12”. Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e. replace “PSS” with “PSS-10” in the whole text."	3	2	life12060901_makarova	0
+Authors introduced an interaction effect long COVID x gender into the model in order to test their third hypothesis (“(iii) whether gender moderated the associations between long 119 COVID status and the health outcomes”, page 3, line 119).			The authors admit that, line 367, younger people were overrepresented, but to me, it was that older persons, especially over age 70, who were somehow missed in the survey sampling process. The study offers little to say about long covid among the elderly.	1	2	life12060901_perova	0
+We have corrected the manuscript according to this guidance.			The authors consider a number of factors associated with long COVID and psychological symptoms. I would suggest another factor - frustration with not getting well. Normally, you get sick, you get better (if you don't die). You get injured moderately and you get better with time. Long COVID breaks that pattern - you get sick and stay sick. Like being sick and tired of being sick and tired. Some uncommon illnesses may act that way, but COVID has become endemic and common. What is your common cold lasted for ten months? People would get really irritated and discouraged by such a situation.	1	2	life12060901_perova	0
+Author Response We thank the reviewers for their guidance on this manuscript.			I accept the manuscript for its publication in this journal.	1	2	life12060901_perova	0
+Again, the authors are overselling their results.			The authors said in the statistic section that “statistical significance was set at p<0.05.” (page 5, line 212). However, later they wrote “The descriptively higher proportion of females with long COVID bordered statistical significance (p=0.05)” page 5, line 230) and “However, the different proportions of men and women with long COVID (22% and 33%, respectively) bordered towards statistical significance […]” (page 10, line 318). As a statistician, I felt very mad about this, because the results speak a different language, this is non-significant result.	1	2	life12060901_perova	0
+The authors of this paper examined whether sociodemographic as well as health-related differences between COVID-19 patients with and without long COVID could be observed.			The authors wrote later in the conclusion section “[…] but a non-significant trend was found for the association between female gender and long COVID.” (page 11, line 389). This is a “no-go” in scientific research, the interpretation of “trends” resulted from non-significant results is strongly misleading. Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05). That is the true story. Hence, I urge the authors not to mislead readers and to re-write these sections.	1	2	life12060901_perova	0
+R3: “[…] men’s perception of poorer health when having long COVID appears to include higher levels of psychological distress, fatigue and perceived stress” (page 11, line 363), Authors’ response: We have revised the sentence; see section 4.4.			The next major issue targets the interpretation of interaction effects. The authors did three models with three different outcomes, namely psychological distress, fatigue, and perceived stress. Authors introduced an interaction effect long COVID x gender into the model in order to test their third hypothesis (“(iii) whether gender moderated the associations between long COVID status and the health outcomes”, page 3, line 119). In the results section, only two (and not three!) interaction effects became significant (psychological distress: p<0.001, fatigue: p<0.05, and perceived stress: p=0.36, see Table 4, page 7, line 269). However, the authors wrote “While participants with long COVID generally perceived more psychological distress, fatigue, and stress than those without long COVID, differences were larger for men than for women” page 9, line 303), Authors’ response:	1	2	life12060901_perova	0
+The authors of this paper examined whether sociodemographic as well as health-related differences between COVID-19 patients with and without long COVID could be observed.			“[…] men’s perception of poorer health when having long COVID appears to include higher levels of psychological distress, fatigue and perceived stress” (page 11, line 363),	1	2	life12060901_perova	0
+As a statistician, I felt very mad about this, because the results speak a different language, this is non-significant result.			and finally “Third, long COVID appears to have a stronger effect on men than on women” (page 11, line 401). Again, the authors are overselling their results. All these statements are not true, only for two out of three outcomes, namely psychological distress and fatigue, but not for perceived stress (see also Figure 3). Hence, I strongly urge the authors to re-write these sentences so that readers are not misled.	1	2	life12060901_perova	0
+Everything read more smoothly and I could follow authors’ argumentations easily, especially the non-significant gender effect and the interpretation of the interaction effects.			Chapter 2 Materials and Methods: Something strange happened to the alignment of heading “2.2 Sample” (page 3, line 128).	1	2	life12060901_perova	0
+Hence, I urge the authors not to mislead readers and to re-write these sections.			Please fix this. Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”). The total sample is reported here as 303. However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310). I know, there were 7 missing values within the 310. However, this table is still wrong, it must be 310 and not 303. The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%). Hence, I urge the authors to re-calculate percentages in Table 1. Otherwise, this is misleading the readership.	1	2	life12060901_perova	0
+Please use a consistent writing style throughout the whole manuscript.			Chapter 2.3 Measures: Sometimes the verbal scales are set in italic (e.g., “0=better than ususal”, page 4, line 173) and sometimes not (e.g., “4=very often”, page 5, line 187).	1	2	life12060901_perova	0
+In my opinion, the revised manuscript increased a lot in comparison to the first version.			Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold. For example, see Table 4 (page 7, line 269): Column Fatgigue, why is ES not printed in bold? Please use a consistent writing style.	1	2	life12060901_perova	0
+I really enjoyed reading this article and thank the authors for the opportunity to review their interesting paper.			Please use a consistent writing style throughout the whole manuscript. Results (page 7, line 274): Sometimes fatigue (page 7, line 274) and sometimes with a capital F (Fatigue, page 7, line 283).	1	2	life12060901_perova	0
+The authors did a cross-sectional online survey with a total sample of 1649 adults across four countries (Norway, UK, USA, and Australia).			Before a final publication of this paper, I have two minor points left: Wrong column headings in tables: I detected that the authors replaced “GHQ” with “psychological distress” in the text (also “PSS” replaced with “perceived stress”). This is correct. Psychological distress is the latent construct which is measured with the instrument GHQ. Unfortunately, the authors missed to replace GHQ by psychological distress in Table 4 (page 7, line 280) and Table 5 (page 7, line 297; see column headings: not “GHQ”, “Fatigue”, and “Perceived stress” but rather “Psychological distress”, “Fatigue”, and “Perceived stress”). Please correct the two tables.	3	2	life12060901_perova	0
+In addition, the authors introduced an interaction effect between ´long COVID (yes/no) and gener (men/women) and found that men reported significantly higher psychological distress and higher fatigue than women but not for perceived stress.			"Wrong use of abbreviations: The authors introduced the abbreviation “GHQ-12” for the 12-item General Health Questionnaire (see page 4, line 161), but wrote in the text only ""GHQ"" (e.g., page 5, line 207; page 6, line 256; page 7, line 274). Please replace every single “GHQ” in the text with the correct abbreviation “GHQ-12”. Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e. replace “PSS” with “PSS-10” in the whole text."	3	2	life12060901_perova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			The literature seems to indicate a low value of the bending modulus (alpha) that corresponds to a persistence length of 500 microns. The authors instead use alpha = 2 × 10−21 Nm2, which is 1000× larger. With this value, the persistence length = alpha/(kbT) = 0.5 metres. This seems orders of magnitude too large, especially in the face of the results of Boal and Ng [44]. The authors need to be in a physical regime of parameter space in order to confront their results with observed patterns. Otherwise, they are just fitting a non-linear phenomenon with a (potentially inappropriate) non-linear model— in this case agreement does not provide firm insight into the physical system.	1	2	life4030433_makarova	0
+I have a few comments, suggestions, and questions.			In Section 3.3 the authors describe how an increase in system size indicated that one of the results presented earlier in the paper (at beta = 0.5) was an artifact of a smaller system size. This brings into question all results at the smaller system size, potentially including all the quantitative results of the paper. The authors should be able to argue that the other results will not depend on system size. The correlation length in Figure 3b is by eye more than the system size, indicating a potential qualitative change with increasing system size (which the authors find). The correlation length in other figures is less than the system size, indicating that they may be fine. The authors need to add some of this discussion to reassure the reader.	1	2	life4030433_makarova	0
+In response to the reviewers’ comments: (1) We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low.			The authors find that results at beta = 0 are most similar to the experimental reticulate patterns. In the force model, there are three forces: elastic, gliding, and contact/cohesion. The contact/cohesion force is presented in equation 12, and is proportional to epsilon. After Equation 23, the authors define beta = epsilon/(zeta⋅v0), implying that there will be no contact/cohesion forces when beta = 0. However, in the results of Figures 3, 4, and 5 the results for beta = 0 clearly show interactions between the filaments. The fourth paragraph of section 2.9 describes a hard-core interaction, which appears to be the source of the patterns when beta = 0. This hard-core interaction should be mentioned after Equation 12, since it is not simply an implementation issue but the dominant interaction with beta = 0.	1	2	life4030433_makarova	0
+Similarly, it is odd that e is a vector and [e] is a matrix.			All the results depend on the cohesion forces, which are described by a Lennard–Jones function in Equation 12, in Section 2.5. However, this way of approximating cohesion is not well justified, and in Section 2.9 (several sections later) the authors mention that in reality the attractive force “does not really exist” and that the trichomes cohere after contact. The authors are using an LJ interaction to approximate contact adhesion. They should say why, and address how good of an approximation they expect this to be.	1	2	life4030433_makarova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			In Section 2.4, it is stated that a stochastic process determines P, but no details are given. There should be some description, if brief, for this process. What distribution is used?	1	2	life4030433_makarova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			The authors mention in the discussion that they may get stronger results for a deeper system, or for trichomes that may only glide freely near a surface or other trichome. In terms of the understanding gained from the modelling/simulations, how would these lead to qualitative changes in the observed patterns?	1	2	life4030433_makarova	0
+Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			The viscosity mu = 1 Pa s is used. This value is approximately 1000 times higher than the viscosity of water, which is the environment of filamentous cyanobacteria. A reference is given, but there should also be a brief justification in the text.	1	2	life4030433_makarova	0
+Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			The authors do not include a description of the reaction force for the top and bottom planes. A brief explanation would be appropriate.	1	2	life4030433_makarova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			None of the references at the end of the paper contain journal volume or page numbers. They should.	1	2	life4030433_makarova	0
+The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display.			In general, and especially in the introduction, many statements are made without accompanying references.	1	2	life4030433_makarova	0
+The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			There are supplemental movies attached. These are not referred to in the text.	1	2	life4030433_makarova	0
+They also make quantitative measurements to describe the behaviour of the model trichomes.			At the end of the discussion of the bending modulus (in Section 2.8, on page 11), the implied persistence length and the irrelevance of thermal fluctuations should be explicitly mentioned.	3	2	life4030433_makarova	0
+The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			In Section 4, the discussion section of the paper, the topic of the bending modulus is returned to for a paragraph. This paragraph needs to explicitly address why the measurements of Boal and Ng were not used, i.e., why Boal and Ng misinterpreted their data. In particular the authors should suggest how the measurements are significant measurements to not use. In particular this paragraph needs to R8 suggest how the measurement should be done to properly test their suggested values for the bending modulus, and so resolve the disagreement with Boal and Ng.	3	2	life4030433_makarova	0
+Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			• ˆ h looks like a unit vector parallel to h. Is there a reason that h only has a ˆ ⋅ in Equation (12)?	1	2	life4030433_makarova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			Walter (ref 7) describes the formation of “clumps” which are very similar to the structures describes by Shepard. Shepard and Sumner provide better images (Walter only shows sketches), but it might be worth mentioning that the phenomenon is more general. As I recall, Petroff et al. (ref 13) show similar patterns.	1	2	life4030433_makarova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			In the force balance, the drag coeffcient is assumed to be the viscosity of water. Since the cells glide over a surface and next to other cells, it would seem that cell–cell friction and cell–surface friction might be at least as important as hydrodynamic drag. What about the viscosity of the material used in the “slime jet”?	1	2	life4030433_makarova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			Following Equation (13), it is stated that “The interpolation ensures the net force and net torque of the interaction forces are null”. This confuses me so I suspect that I have misunderstood something. In an overdamped system, such as this one, all forces and torques are balanced by drag on the cell. Is this all that is happening in this derivation? The text reads like certain forces and torques are assumed to vanish. Are these forces and torques introduced by the interpolation and the parameterization is chosen to cancel artifacts? Or, are there internal forces and and torques acting on the vertices that cancel as a result of Newton’s second law? Please clarify this point.	1	2	life4030433_makarova	0
+Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			In the estimation of the Young’s modulus it is assumed that the bending resistance is due to the cell wall. I suspect that this is a very poor approximation, accounting for the two order of magnitude difference between the estimated and benchmark values of Y. It would seem that the resistance is controlled or at least influenced by additional factors such as protein expression (notably MreB) and osmotic pressure. If my intuition is wrong (I am not an expert on cell morphology), I would appreciate knowing why.	1	2	life4030433_makarova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			If I understand correctly, there are six important length scales in the problem, the size of the domain DH , the spacing between trichomes 13 ρ − , 0 , , L l αν ζ = Θ, and the trichome length L = Nl (also denoted L in the manuscript), giving five dimensionless numbers. However, in the non-dimensionalization, the authors state that there are only two parameters: ρ and Nl γ θ = . Unless I am mistaken, there are additional parameters like l/L, Nl/D, and N that the authors hold fixed but may influence the details of the numerics.	1	2	life4030433_makarova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			It seems a little odd to posit an attractive force in the derivation (Equation (12)) and then ignore/dismiss it in the numerics (“… in reality the attractive force does not really exist, trichomes only cohere after they have come into contact” page 12). Clearly, this is done to (quite reasonably) avoid modeling steric interactions with hard bodies. It was a little distracting to think that sticky adhesion was decaying like 6 ( ) h Θ . Perhaps note after Equation (12) that the attractive force decreases to zero at a finite distance in the numerics.	1	2	life4030433_makarova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			Can you coarse grain this model to make it analytically tractable? The basic phenomena suggest something like an active shear-thinning uid? This would make it easier to understand the bifurcations in the dynamics and may require fewer parameters. Just something to think about.	1	2	life4030433_makarova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			Notably, the introduction of reproduction does not introduce another parameter. As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+). Thus, the length of a filament is not a model parameter. Rather, length is determined dynamically as ~ N k τ + (10)	1	2	life4030433_makarova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			This project would benefit immeasurably from a quantitative comparison to experiments. I understand that this is beyond the scope of this paper, but I strongly encourage future work in this direction.	1	2	life4030433_makarova	0
+The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			In the introduction, this project is put into the context of stromatolites. In the discussion/conclusion, I would appreciate to hear the authors thoughts on what this model teaches us about stromatolites. In the current form I am left to conclude only that communities of filamentous bacteria can develop reticulated patterns similar to forms observed in the fossil record. This conclusion does not offer much more than Shepard’s experimental observation of this result. Can we use the details of this model to, for example, estimate the cohesion β of ancient cells? Do these results provide any insight into the identity of stromatolite building cells. As far as I can tell, nothing in this model suggests that photosynthesis (much less oxygenic photosynthesis) is required to form these patterns. It seems that this model shows that polygonal patters are a generic feature of elongated gliding cells and is independent of metabolism. A quick google search produced this video of mat of the sulfur-oxidizing bacteria Beggiatoa spp. http://vimeo.com/57205513 (note: I have no idea of the details of these observations so any similarity may be spurious).	1	2	life4030433_makarova	0
+However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			The literature seems to indicate a low value of the bending modulus (alpha) that corresponds to a persistence length of 500 microns. The authors instead use alpha = 2 × 10−21 Nm2, which is 1000× larger. With this value, the persistence length = alpha/(kbT) = 0.5 metres. This seems orders of magnitude too large, especially in the face of the results of Boal and Ng [44]. The authors need to be in a physical regime of parameter space in order to confront their results with observed patterns. Otherwise, they are just fitting a non-linear phenomenon with a (potentially inappropriate) non-linear model— in this case agreement does not provide firm insight into the physical system.	1	2	life4030433_perova	0
+Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			In Section 3.3 the authors describe how an increase in system size indicated that one of the results presented earlier in the paper (at beta = 0.5) was an artifact of a smaller system size. This brings into question all results at the smaller system size, potentially including all the quantitative results of the paper. The authors should be able to argue that the other results will not depend on system size. The correlation length in Figure 3b is by eye more than the system size, indicating a potential qualitative change with increasing system size (which the authors find). The correlation length in other figures is less than the system size, indicating that they may be fine. The authors need to add some of this discussion to reassure the reader.	1	2	life4030433_perova	0
+The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			The authors find that results at beta = 0 are most similar to the experimental reticulate patterns. In the force model, there are three forces: elastic, gliding, and contact/cohesion. The contact/cohesion force is presented in equation 12, and is proportional to epsilon. After Equation 23, the authors define beta = epsilon/(zeta⋅v0), implying that there will be no contact/cohesion forces when beta = 0. However, in the results of Figures 3, 4, and 5 the results for beta = 0 clearly show interactions between the filaments. The fourth paragraph of section 2.9 describes a hard-core interaction, which appears to be the source of the patterns when beta = 0. This hard-core interaction should be mentioned after Equation 12, since it is not simply an implementation issue but the dominant interaction with beta = 0.	1	2	life4030433_perova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			All the results depend on the cohesion forces, which are described by a Lennard–Jones function in Equation 12, in Section 2.5. However, this way of approximating cohesion is not well justified, and in Section 2.9 (several sections later) the authors mention that in reality the attractive force “does not really exist” and that the trichomes cohere after contact. The authors are using an LJ interaction to approximate contact adhesion. They should say why, and address how good of an approximation they expect this to be.	1	2	life4030433_perova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			In Section 2.4, it is stated that a stochastic process determines P, but no details are given. There should be some description, if brief, for this process. What distribution is used?	1	2	life4030433_perova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			The authors mention in the discussion that they may get stronger results for a deeper system, or for trichomes that may only glide freely near a surface or other trichome. In terms of the understanding gained from the modelling/simulations, how would these lead to qualitative changes in the observed patterns?	1	2	life4030433_perova	0
+Notably, the introduction of reproduction does not introduce another parameter.			The viscosity mu = 1 Pa s is used. This value is approximately 1000 times higher than the viscosity of water, which is the environment of filamentous cyanobacteria. A reference is given, but there should also be a brief justification in the text.	1	2	life4030433_perova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			The authors do not include a description of the reaction force for the top and bottom planes. A brief explanation would be appropriate.	1	2	life4030433_perova	0
+The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			None of the references at the end of the paper contain journal volume or page numbers. They should.	1	2	life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			In general, and especially in the introduction, many statements are made without accompanying references.	1	2	life4030433_perova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			There are supplemental movies attached. These are not referred to in the text.	1	2	life4030433_perova	0
+Rather, length is determined dynamically as ~ N k τ + (10)			At the end of the discussion of the bending modulus (in Section 2.8, on page 11), the implied persistence length and the irrelevance of thermal fluctuations should be explicitly mentioned.	3	2	life4030433_perova	0
+Round 2: Author Response to Reviewer 1 We have addressed the reviwer’s requests: (1)			In Section 4, the discussion section of the paper, the topic of the bending modulus is returned to for a paragraph. This paragraph needs to explicitly address why the measurements of Boal and Ng were not used, i.e., why Boal and Ng misinterpreted their data. In particular the authors should suggest how the measurements are significant measurements to not use. In particular this paragraph needs to R8 suggest how the measurement should be done to properly test their suggested values for the bending modulus, and so resolve the disagreement with Boal and Ng.	3	2	life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			Notation: • L is used both for the length of the trichome Nl and in the non-dimensionalization as 0 αν ζ . • I have never seen , ⋅⋅used to define an edge. It looks like an inner product. Is this standard? • In Equations (5)–(8), is there any difference between subscripts and superscripts? • It is confusing that b κ is a vector that is not simply related to b. I initially thought that κ was a new parameter. Similarly, it is odd that e is a vector and [e] is a matrix. Why not use the Levi–Civita tensor? • ˆ h looks like a unit vector parallel to h. Is there a reason that h only has a ˆ ⋅ in Equation (12)?	1	2	life4030433_perova	0
+While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			Walter (ref 7) describes the formation of “clumps” which are very similar to the structures describes by Shepard. Shepard and Sumner provide better images (Walter only shows sketches), but it might be worth mentioning that the phenomenon is more general. As I recall, Petroff et al. (ref 13) show similar patterns.	1	2	life4030433_perova	0
+However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			In the force balance, the drag coeffcient is assumed to be the viscosity of water. Since the cells glide over a surface and next to other cells, it would seem that cell–cell friction and cell–surface friction might be at least as important as hydrodynamic drag. What about the viscosity of the material used in the “slime jet”?	1	2	life4030433_perova	0
+Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			Following Equation (13), it is stated that “The interpolation ensures the net force and net torque of the interaction forces are null”. This confuses me so I suspect that I have misunderstood something. In an overdamped system, such as this one, all forces and torques are balanced by drag on the cell. Is this all that is happening in this derivation? The text reads like certain forces and torques are assumed to vanish. Are these forces and torques introduced by the interpolation and the parameterization is chosen to cancel artifacts? Or, are there internal forces and and torques acting on the vertices that cancel as a result of Newton’s second law? Please clarify this point.	1	2	life4030433_perova	0
+As a result, I do not recommend that this paper should be published in its current form.			In the estimation of the Young’s modulus it is assumed that the bending resistance is due to the cell wall. I suspect that this is a very poor approximation, accounting for the two order of magnitude difference between the estimated and benchmark values of Y. It would seem that the resistance is controlled or at least influenced by additional factors such as protein expression (notably MreB) and osmotic pressure. If my intuition is wrong (I am not an expert on cell morphology), I would appreciate knowing why.	1	2	life4030433_perova	0
+Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			If I understand correctly, there are six important length scales in the problem, the size of the domain DH , the spacing between trichomes 13 ρ − , 0 , , L l αν ζ = Θ, and the trichome length L = Nl (also denoted L in the manuscript), giving five dimensionless numbers. However, in the non-dimensionalization, the authors state that there are only two parameters: ρ and Nl γ θ = . Unless I am mistaken, there are additional parameters like l/L, Nl/D, and N that the authors hold fixed but may influence the details of the numerics.	1	2	life4030433_perova	0
+Rather, length is determined dynamically as ~ N k τ + (10)			It seems a little odd to posit an attractive force in the derivation (Equation (12)) and then ignore/dismiss it in the numerics (“… in reality the attractive force does not really exist, trichomes only cohere after they have come into contact” page 12). Clearly, this is done to (quite reasonably) avoid modeling steric interactions with hard bodies. It was a little distracting to think that sticky adhesion was decaying like 6 ( ) h Θ . Perhaps note after Equation (12) that the attractive force decreases to zero at a finite distance in the numerics.	1	2	life4030433_perova	0
+They also explore behaviour in much of the available parameter space.			The parameter space is rather large, and only a small subspace is explored in the numerics. This is fine, but it leaves a lot of interesting questions un-addressed. For example, is the transition from broad streams (Figure 4a) to loops (Figure 4d) continuous or are there bifurcations/phase transitions? Can you coarse grain this model to make it analytically tractable? The basic phenomena suggest something like an active shear-thinning uid? This would make it easier to understand the bifurcations in the dynamics and may require fewer parameters. Just something to think about.	1	2	life4030433_perova	0
+Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			In this model, the density of cells is assumed to be constant. However, the timescale of the simulations is comparable to the doubling time of many bacteria. Because cell density is found to R6 be important in determining the morphology of communities, I suspect that reproduction is important in understanding the patterns. This point might be worth addressing in the text.	1	2	life4030433_perova	0
+However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			This project would benefit immeasurably from a quantitative comparison to experiments. I understand that this is beyond the scope of this paper, but I strongly encourage future work in this direction.	1	2	life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			In the introduction, this project is put into the context of stromatolites. In the discussion/conclusion, I would appreciate to hear the authors thoughts on what this model teaches us about stromatolites. In the current form I am left to conclude only that communities of filamentous bacteria can develop reticulated patterns similar to forms observed in the fossil record. This conclusion does not offer much more than Shepard’s experimental observation of this result. Can we use the details of this model to, for example, estimate the cohesion β of ancient cells? Do these results provide any insight into the identity of stromatolite building cells. As far as I can tell, nothing in this model suggests that photosynthesis (much less oxygenic photosynthesis) is required to form these patterns. It seems that this model shows that polygonal patters are a generic feature of elongated gliding cells and is independent of metabolism. A quick google search produced this video of mat of the sulfur-oxidizing bacteria Beggiatoa spp. http://vimeo.com/57205513 (note: I have no idea of the details of these observations so any similarity may be spurious).	1	2	life4030433_perova	0
+The text was revised according to the comments.			As depicted in some of the electron micrographs in Figure 3, the authors describe the presence of thin filaments within the sheaths surrounding the motile filaments. Is it possible that these are in fact type IV pili? How does the structure compare to other published reports of type IV pili?	1	2	life4040819_makarova	0
+R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			In Section 3.5, Analysis of Movement of Filaments, the descriptions were confusing at times. For instance Line 253 states “only in some exceptional cases (the straight line in Figure 6D), net displacement was achieved”. Net displacement presumably refers to any final position after a cycle that differs from the starting position. In other words, net displacement would refer to any value other than zero in Figure 6D. All of the points on this graph appear to indicate net displacement, albeit some less than others. Do the authors mean to state that only in a few instances was a large net displacement achieved? Otherwise the authors’ statement does not appear correct. Perhaps using numerical values instead of statements such as “the average velocity was quite limited” would help to clarify.	1	2	life4040819_makarova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			The authors observe that the velocity of movement was maximal immediately after a reversal and then declined exponentially until the next reversal. The authors then suggest that this observation is theoretically inconsistent with type IV pili or focal adhesion complexes as the driving force for movement. Are there any available published reports quantifying the motility of organisms using these systems that would corroborate this empirically? In short, has it been demonstrated that bacteria moving by twitching motility or focal adhesion do not show a similar pattern for velocity of movement?	1	2	life4040819_makarova	0
+A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			The authors state that supracellular structures emerge from unordered motion and are therefore not encoded for genetically. While their model for spiral formation does not require a genetic program for controlling motility, it may not always be the case for other supercellular structures. In other words, it may not always be true that supercellular structures are not genetically determined.	1	2	life4040819_makarova	0
+Response: We thank the reviewers for giving us helpful comments.			Line 45—suggest adding “to” before “back-and-forth motion...”	1	2	life4040819_makarova	0
+Using TEM, the authors observed junctional pore complexes arrayed at septa, and extending through the cell envelope to the exterior of the cell, and potentially identified basal body complexes at the cytoplasmic face of the inner membrane.			Lines 268 and 269—“(D)” and “(E)” should be bolded to be consistent	1	2	life4040819_makarova	0
+Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			Line 287—appears to be missing e-values. The beginning of this sentence indicates 10 genes and then only gives 8 respective e-values for the 10 genes.	1	2	life4040819_makarova	0
+Response: We thank the reviewers for giving us helpful comments.			Line 290—do orf303 and orf297 have homology to any characterized proteins? Are they conserved hypotheticals? Is there any indication of the function for these?	1	2	life4040819_makarova	0
+Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			Line 415—“single” appears to be formatted differently than the rest of the text	1	2	life4040819_makarova	0
+The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			About 20 years ago it was shown that in order to preserve crucial ultrastructural features of cyanobacterial cell walls, the cells have to be processed using cryo-procedures (Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395). The authors use an outdated and very artifact-laden method to process their samples. This choice is responsible for a number of shortcomings that need to be addressed: The lack of certain features such as the S-layer and the oscillin fibrils in the reported pictures are attributable to the choice of preparation, as they are clearly present in two species of the same genus described in the above cited paper. Moreover, these structures would explain the rotation of the cells that now is somewhat “mysteriously” explained by a helical slime ejection that is not supported by the observed arrangement of the nozzles. Therefore the authors should do the following: Response:	1	2	life4040819_makarova	0
+In addition, Figure 3A was replaced by a figure of better contrast.			(a) Clearly spell out in the text why there are no oscillin fibrils and surface structures visible and compare their results with the published results of the cell wall structure of the species of the same genus prepared using cryo-procedures.	1	2	life4040819_makarova	0
+Response: We thank the reviewers for giving us helpful comments.			(b) There is no clear distinction in the text between the “sheath” and the secreted “slime” of the cells, which are two completely different structures both physically and chemically (Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Figure 1A shows a filament that lacks the “sheath”, while in other pictures the structure is visible. The reason for this is the following: cyanobacteria of the genus Phormidium build over time a carbohydrate layer on their surface that is physically attached to their cell surface, usually called the sheath. Filaments that are ensheathed are non-motile! In contrast, the slime that is secreted by gliding filaments of the same species is not visible in TEM preparations, even when cryo-preservation methods are used (see Figure 1A of the ms and Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395 and Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Therefore, the authors should go through their ms and make a careful distinction between these two structures. For example, in the discussion it sounds as if the “sheath” is preventing the R5 cells from sliding, however this is the slime tube that is secreted that is not physically attached to the cell surface at all.	1	2	life4040819_makarova	0
+The text was revised according to the comments.			(c) Please omit the description of the basal body part of the junctional pores or provide substantially better pictures. Even after careful inspection of your images, I cannot convince myself of seeing these parts. Moreover, in cryo-preserved and freeze-fractured cells there are no such structures visible. That does not mean that they don’t exist, it may only mean that none of the so far used preservation methods reveals these structures.	1	2	life4040819_makarova	0
+Using TEM, the authors observed junctional pore complexes arrayed at septa, and extending through the cell envelope to the exterior of the cell, and potentially identified basal body complexes at the cytoplasmic face of the inner membrane.			(d) Please omit the description about helical tilted junctional pores. These structures are clearly not arranged in this way and the reasons for rotation are the unfortunately not-preserved surface structures mentioned above.	1	2	life4040819_makarova	0
+A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			Results section headings and figure captions should be phrased as conclusions, supported by the data presented, when appropriate (can be a minor change, for example 3.2 could be titled, “Phormidium colonies form divergent structures,” or larger, for example 3.5 could be titled, “Individual filaments undergo directional reversals, with maximum velocity immediately following a reversal.”	1	2	life4040819_makarova	0
+The observations and models are consistent with previously published work on motility of various cyanobacteria and provide new insights into the mechanics of motility of cyanobacteria that are interesting.			In the discussion there are further points that need to be addressed: (a) Please clarify the source of the helical flow of the slime, which is most likely due to the presence of the not-preserved helically arranged cell surface proteins not the arrangement or tilt of the junctional pores.	1	2	life4040819_makarova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			(b) In the model that describes the potential activity of the junctional pores (How myxobacteria glide. Curr. Biol. 2002, 12, 369–377) it is not suggested that the nozzle actually “contracts” as mentioned in the ms. The swelling of the slime material in the nozzle fills the nozzle and eventually will generate a counter-force of the nozzle walls of the nozzle that results in the ejection of the slime. Please rephrase the text accordingly.	1	2	life4040819_makarova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			(c) When the cells form spirals, there are two components to this process: the formation of spirals and their macroscopic sense of rotation: clock- or counterclockwise. In the discussion it is assumed that the rotation of the filaments is crucial for both of these components. If this would be true, then no spirals should be observed in non-rotating filamentous gliding cyanobacteria. However, this is not the case. Anabaena spec. a non-rotating species can generate spirals (see Figure X; Mucilage secretion and the movements of blue-green-algae. Protoplasm 1968, 65, 223–238). This means that the sense of rotation may only be responsible for the direction of the spiral but not for its emergence in the first place. It would be good if the authors would distinguish between these two phenomena and discuss them accordingly.	1	2	life4040819_makarova	0
+Within the text, corrected words are highlighted.			(d) In Lines 309–315: References 17 and 22 are accurately described, however these models relate to gliding in Myxobacteria, a distinction not noted in the text. Thus, comparisons between these models (including the focal adhesion model) and the current model are only valid if the mechanism of motility in these microorganisms is the same. The authors should (briefly) explain their reasoning on the validity of comparing these two systems, and the implications of their findings to motility in Myxobacteria.	1	2	life4040819_makarova	0
+The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			Lines 94–95: Is it standard to sequence an entire genome, but only deposit several clusters, or should the entire genome be accessible to allow other researchers access to this source?	1	2	life4040819_makarova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			Lines 227–230: The interpretation of the counterclockwise spirals here is somewhat distracting. It can be left as an observation here, and the model explained later.	1	2	life4040819_makarova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			Lines 316–317: This model for slime secretion suggests that rather than slime being comprised of a single, long, polysaccharide, it is made of smaller subunits that are secreted in a step-wise fashion. The authors should connect this idea to the finding that pseudopilins appear to be part of the molecular machinery, as the model and the data are coherent on this point.	1	2	life4040819_makarova	0
+This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.	1	2	life4040819_perova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			As depicted in some of the electron micrographs in Figure 3, the authors describe the presence of thin filaments within the sheaths surrounding the motile filaments. Is it possible that these are in fact type IV pili? How does the structure compare to other published reports of type IV pili?	1	2	life4040819_perova	0
+(3) In the discussion there are further points that need to be addressed: (a)			In Section 3.5, Analysis of Movement of Filaments, the descriptions were confusing at times. For instance Line 253 states “only in some exceptional cases (the straight line in Figure 6D), net displacement was achieved”. Net displacement presumably refers to any final position after a cycle that differs from the starting position. In other words, net displacement would refer to any value other than zero in Figure 6D. All of the points on this graph appear to indicate net displacement, albeit some less than others. Do the authors mean to state that only in a few instances was a large net displacement achieved? Otherwise the authors’ statement does not appear correct. Perhaps using numerical values instead of statements such as “the average velocity was quite limited” would help to clarify.	1	2	life4040819_perova	0
+In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			The authors observe that the velocity of movement was maximal immediately after a reversal and then declined exponentially until the next reversal. The authors then suggest that this observation is theoretically inconsistent with type IV pili or focal adhesion complexes as the driving force for movement. Are there any available published reports quantifying the motility of organisms using these systems that would corroborate this empirically? In short, has it been demonstrated that bacteria moving by twitching motility or focal adhesion do not show a similar pattern for velocity of movement?	1	2	life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			The authors state that supracellular structures emerge from unordered motion and are therefore not encoded for genetically. While their model for spiral formation does not require a genetic program for controlling motility, it may not always be the case for other supercellular structures. In other words, it may not always be true that supercellular structures are not genetically determined.	1	2	life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			Line 45—suggest adding “to” before “back-and-forth motion...”	1	2	life4040819_perova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			Lines 268 and 269—“(D)” and “(E)” should be bolded to be consistent	1	2	life4040819_perova	0
+R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			Line 287—appears to be missing e-values. The beginning of this sentence indicates 10 genes and then only gives 8 respective e-values for the 10 genes.	1	2	life4040819_perova	0
+In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			Line 290—do orf303 and orf297 have homology to any characterized proteins? Are they conserved hypotheticals? Is there any indication of the function for these?	1	2	life4040819_perova	0
+R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			Line 415—“single” appears to be formatted differently than the rest of the text	1	2	life4040819_perova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			About 20 years ago it was shown that in order to preserve crucial ultrastructural features of cyanobacterial cell walls, the cells have to be processed using cryo-procedures (Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395). The authors use an outdated and very artifact-laden method to process their samples. This choice is responsible for a number of shortcomings that need to be addressed: The lack of certain features such as the S-layer and the oscillin fibrils in the reported pictures are attributable to the choice of preparation, as they are clearly present in two species of the same genus described in the above cited paper. Moreover, these structures would explain the rotation of the cells that now is somewhat “mysteriously” explained by a helical slime ejection that is not supported by the observed arrangement of the nozzles. Therefore the authors should do the following: Response:	1	2	life4040819_perova	0
+Round 1: and Author Response The manuscript “Cellular dynamics drives emergence of supracellular structure in the cyanobacterium Phormidium sp.			Clearly spell out in the text why there are no oscillin fibrils and surface structures visible and compare their results with the published results of the cell wall structure of the species of the same genus prepared using cryo-procedures.	1	2	life4040819_perova	0
+Within the text, corrected words are highlighted.			The quality of Figure 1A was rather bad. The contrast was corrected and a new figure was inserted. In the new Figure 1A, the sheath is quite visible. The quality of EM figure is very low as embedded in an MS word file. We provide a better quality figure as supplemental Figure S1. The use of “sheath” is therefore correct in the paragraph describing the figure. In the model, the word “sliding” was not understood in the sense in which we wanted it to be. This is lateral sliding that provokes curvature of the filament. This point was clarified in the text.	1	2	life4040819_perova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			Please omit the description of the basal body part of the junctional pores or provide substantially better pictures. Even after careful inspection of your images, I cannot convince myself of seeing these parts. Moreover, in cryo-preserved and freeze-fractured cells there are no such structures visible. That does not mean that they don’t exist, it may only mean that none of the so far used preservation methods reveals these structures.	1	2	life4040819_perova	0
+Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			Please omit the description about helical tilted junctional pores. These structures are clearly not arranged in this way and the reasons for rotation are the unfortunately not-preserved surface structures mentioned above.	1	2	life4040819_perova	0
+The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			Results section headings and figure captions should be phrased as conclusions, supported by the data presented, when appropriate (can be a minor change, for example 3.2 could be titled, “Phormidium colonies form divergent structures,” or larger, for example 3.5 could be titled, “Individual filaments undergo directional reversals, with maximum velocity immediately following a reversal.”	1	2	life4040819_perova	0
+The contribution of motility to the formation of supracellular structure is generally an area of research regarding cyanobacteria that has been overlooked.			Please clarify the source of the helical flow of the slime, which is most likely due to the presence of the not-preserved helically arranged cell surface proteins not the arrangement or tilt of the junctional pores.	1	2	life4040819_perova	0
+This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			In the model that describes the potential activity of the junctional pores (How myxobacteria glide. Curr. Biol. 2002, 12, 369–377) it is not suggested that the nozzle actually “contracts” as mentioned in the ms. The swelling of the slime material in the nozzle fills the nozzle and eventually will generate a counter-force of the nozzle walls of the nozzle that results in the ejection of the slime. Please rephrase the text accordingly.	1	2	life4040819_perova	0
+The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			When the cells form spirals, there are two components to this process: the formation of spirals and their macroscopic sense of rotation: clock- or counterclockwise. In the discussion it is assumed that the rotation of the filaments is crucial for both of these components. If this would be true, then no spirals should be observed in non-rotating filamentous gliding cyanobacteria. However, this is not the case. Anabaena spec. a non-rotating species can generate spirals (see Figure X; Mucilage secretion and the movements of blue-green-algae. Protoplasm 1968, 65, 223–238). This means that the sense of rotation may only be responsible for the direction of the spiral but not for its emergence in the first place. It would be good if the authors would distinguish between these two phenomena and discuss them accordingly.	1	2	life4040819_perova	0
+Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			In Lines 309–315: References 17 and 22 are accurately described, however these models relate to gliding in Myxobacteria, a distinction not noted in the text. Thus, comparisons between these models (including the focal adhesion model) and the current model are only valid if the mechanism of motility in these microorganisms is the same. The authors should (briefly) explain their reasoning on the validity of comparing these two systems, and the implications of their findings to motility in Myxobacteria.	1	2	life4040819_perova	0
+R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			Lines 94–95: Is it standard to sequence an entire genome, but only deposit several clusters, or should the entire genome be accessible to allow other researchers access to this source?	1	2	life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			Lines 227–230: The interpretation of the counterclockwise spirals here is somewhat distracting. It can be left as an observation here, and the model explained later.	1	2	life4040819_perova	0
+The text was revised according to the comments.			Lines 316–317: This model for slime secretion suggests that rather than slime being comprised of a single, long, polysaccharide, it is made of smaller subunits that are secreted in a step-wise fashion. The authors should connect this idea to the finding that pseudopilins appear to be part of the molecular machinery, as the model and the data are coherent on this point.	1	2	life4040819_perova	0
+I have only a few comments, which might improve the manuscript.			Section 1. The place of RNA in LUCA (page 2): In search of features that are more conserved (carrying deep phylogenetic memory) than the sequence of genes, Wächtershäuser focuses on a paper of his in Systematic and Applied Microbiology (1998) that uses gene content and order of microbial genomes to make inferences about the last universal common ancestor (LUCA) of cellular life. He then mines the significance of some of the conserved chromosomal segments in light of some other evidence. The exercise is at places compelling, but forgets some recent, very global and exhaustive analyses that also use highly conserved biological features to reconstruct the makeup of LUCA (e.g., gene order, 3D molecular structure, molecular functions). See for example PubMed references PMID: 17370266, PMID: 21612591, and PMID: 17908824, which are in line with some conclusions derived from the alignment of Figure 1. The fact that these other analyses make use of hundreds of genomes to infer the ancient biochemistry of LUCA complements and strengthens the preliminary and fragmentary analysis of only 19 of them by the author, which also excludes eukaryotic genomes from the set (understandably, few sequenced genomes were available in 1998, and eukaryotes are in general “master rearrangers”).	1	2	life4041050_makarova	0
+In light of recent research developments and argumentation (some of it reviewed), his views should be considered a welcome addition to the many ideas that populate the “origin of life” field of inquiry that counter the dominant paradigm.			Given my interest in the bioinformatics of gene content and order, I took the liberty of studying the brief Systematic & Applied Microbiology paper to check the validity of the “reconstruction” methods of Figure 1. The algorithmic implementation that is described is quite raw and does not extract important information that is embedded in the clusters of conserved gene segments. Important algorithms have been devised since the initial work of Sankoff in the sixties and seventies to do exactly that. I refer to the work of Pevzner, Tesler and Bourque as good examples, but also of Warnow. I also recommend visiting GRIMM (http://grimm.ucsd.edu/GRIMM/) and perhaps using the server to confirm or extract additional information from the alignments that are summarized in Figure 1. The Wächtershäuser algorithm makes use of conserved elements of gene content and order but discards information provided by the actual rearrangement operations that erase gene order history. The algorithm does not describe how gene homology was detected, how the limited set of genes was selected, and how the alignments were constructed (I imagine by hand). It is not clear if a guiding tree was used in the alignment (though this is mentioned in Line 65), since this is not made explicit in the 1998 publication. An alignment implies a tree but usually alignment algorithms are greedy and problematic and represent the most important limiting step of a phylogeny (the field is thus moving to the joint alignment and tree reconstruction). Therefore, I do not think the ancestors were properly reconstructed (the tracing of features in ancestor nodes of trees are not described, nor the actual trees). Regardless of all of these limitations, the tight conservation of certain segments is enough to show the existence of a core of ribosomal proteins that is universally present in cellular organisms. This in itself is valuable. Of course, a sample of 19 microbes may not be enough to encompass molecular diversity and the absence of Eukarya may also be problematic for any global evolutionary statement. In other words, the risks of sampling bias are clearly present and should be mentioned in the manuscript.	1	2	life4041050_makarova	0
+He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			The list of common genes in the alignment of Table 1 is enriched in small and large subunit ribosomal proteins that are the most ancient of the ribosomal set, according to the Caetano-Anolles theorem that is mentioned later in the manuscript. Interestingly, the most ancient of them are clustered toward the 5’ end of the genomic sequences that were aligned (S12, S17, S5, S4 and L2, L3 and L24). Could this imply a possible ancient segmental duplication? Also interesting is the placement of the most ancient ribosomal protein S12 between exactly two polymerase (beta and beta prime) and elongation factor (entry and translocation) genes (separated by single and much more derived ribosomal proteins). Could this be an ancient memory of the ribosome mediating translation and replication? A commentary would enhance the value of the section.	1	2	life4041050_makarova	0
+He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			In terms of the genome organization of LUCA (Line 105), there are numerous arguments in favor of an RNA ancestral genome and the late unfolding of DNA as genetic repository (perhaps through viruses). I cannot understand how 2'-hydroxy groups could destabilize the molecule and lead to “intramolecular self-destruction” (Line 107). RNA has the wonderful property of folding in search of energetic and kinetic minima. These processes make use of a frustrated landscape, which is powerful in terms of its biological potential (stability, function, information). It is much more versatile than the rigid DNA alternative, which is enriched in the ‘information’ capacity. None of them have “self destruction” properties, which would have been weeded out by selection and self-organization very early in “chemical” evolution. In turn, everything about LUCA should be considered quite modern and far away from an initial FeS world. I refer to the work of Daniel Lundin and colleagues in Sweden about the rather late rise of some structures of the ribonuceotide reductase enzymes needed to built the DNA polymers. The Maurel theorem stands if one thinks of single genome molecules. But what if there were many short RNA genomes, as anticipated by Woese, each perhaps linked to different and ancient tRNA-like cofactors? As the author mentions, LUCA is contemporary to a rather complex biochemistry, with numerous structures harboring a multitude of active and allosteric sites in proteins, most of which drive central enzymatic functions.	1	2	life4041050_makarova	0
+He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			In Line 114, the role of chromosomal rearrangement in evolution is not well described. What are the effects of genome rearrangements? Several scenarios are possible, including: (1) Genes remain linked R4 probably because it is more efficient to transcribe genes that produce interacting proteins (part of complexes) than those that are not; (2) Genes remain linked if they are of relatively recent origin (rearrangement has not had the change to split them apart); (3) Genes remain linked if they are part of functional groups historically united by genomic regions (encoding metabolic functions or rRNA? ); this includes genes sharing an operon structure for economy purposes in highly reduced organisms such as bacteria; (4) Genes remain linked because they originated when genomic rearrangements were not biochemically motivated and their sequence makeup was later refractory to rearrangement hotspots. There is a rich literature about rearrangement, hotspots and many processes related to these (including domain organization in proteins). Some discussion in this front could be clarifying.	1	2	life4041050_makarova	0
+In light of recent research developments and argumentation (some of it reviewed), his views should be considered a welcome addition to the many ideas that populate the “origin of life” field of inquiry that counter the dominant paradigm.			In relation to comments of Line 188 onwards, Di Giulio, Caetano-Anolles and others have suggested that the genetic code started to unfold prior to LUCA but continue to do so once life diversified. The corollary, is that the complete canonical set of amino acids may have not been encoded in LUCA. In terms of metabolic and biochemical competency, there are numerous and interesting studies, which have not been cited (especially in origins and evolution of modern metabolism). This includes the coevolution theory of the genetic code, the coexistence of prebiotic chemistries with modern metabolic reactions (recently studied for nucleotide metabolism), and theories about the origin of translation.	1	2	life4041050_makarova	0
+The article demonstrates that the iron-sulfur surface metabolism theory at high temperatures has a much better explanatory power than the RNA World theory.			Section 2: Thermal course of evolution (Page 6): Why a focus on thermal energy? The framework should be on thermodynamics, energy dissipation and information, all of which are linked. What is thermally upward or downward adaptation? Is it conquering niches on Earth or a process involving molecular makeup? Vocabulary and definitions are murky, especially related to the links of environmental thermal fluctuations, energy of folding and stability of polymers. Subsection (1) must be rewritten to help the reader understand the ideas. Should all adaptations comply with maximizing energy R5 dissipation? How can this be reconciled with upward and downward trends? No references are provided despite the rich literature underscoring the controversial link between physics, information theory and biology. If the focus is conquering planetary niches, then perhaps reference the contrasting views and some of their proponents (deep sea versus surface; thermophilic versus mesophilic, etc.). Further elaboration of the Wolfenden theorem appears relevant and its connections to what is known about the origin and evolution of metabolism also of importance.	1	2	life4041050_makarova	0
+I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			Section 5. Place of RNA in the origin and early evolution of translation/Pre-translational coding of peptides. The view that is presented in this crucial segment posits a pre-translational mechanism (side-by-side tRNA mediated coding) that very much resembles the ribosomal entropic system, which could be very advanced. What if the pre-translational coding was assembly line-like and mediated by the aminoacyl-tRNA synthetases themselves? See PMID: 23991065 for one such alternative and a related previous model (PMID: 22210458). This would also match commentary of a possible early origin of non-ribosomal peptide synthesis machinery compared to that of the ribosome.	1	2	life4041050_makarova	0
+He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			Section 9. General overview. While the grand finale “Nothing in evolution makes sense except in the light of chemical predetermination” is impressive and summarizes the championing work of the author, chemical predetermination may apply to the very early stages of evolution and not to many of the stages described in the manuscript. How much chemical predetermination can there be with polymers as these explore a minute fraction of the space they make possible? In the enthalpic-entropic gradient that is proposed, there is also a gradient from “predetermination” to historical contingency. The boundaries of such a gradient is murky and the final statement may not apply to much of modern biochemistry, once proteins start to achieve stable complex structure and much earlier that the time of LUCA. In fact, the genome rearrangements that underlie Figure 1 are testament to the historical contingencies that were already at play in LUCA and not to chemical predetermination.	1	2	life4041050_makarova	0
+I have only a few comments, which might improve the manuscript.			Line 406. “LUCA protein cycle”. Why bring LUCA to a link with the proposed “protein cycle”. LUCA is just the last of a chain of ancestor of diversified life. If the protein cycle requires a coupling of the enthalpic activation of amino acids and the entropic biosynthetic action of a primordial ribosome, then perhaps it is better to call it “ribocellular protein cycle”, since this coupling marks the start of modern cells, not necessarily “modern diversified cells” (i.e., LUCA).	1	2	life4041050_makarova	0
+I must note that a careful evaluation of all facets requires expertise in a multitude of disciplines (from prebiotic chemistry and structural biology to evolutionary bioinformatics and biochemistry) and considerable time, none of which I possess.			Please explain what do you mean with Kandler, Wolfenden etc. Theorem. Is a theorem Kandler’s theory or proposal?	1	2	life4041050_makarova	0
+Round 1: and Author Response The manuscript comprises an exciting account on the origin of life with emphasis on the emergence and function of RNA.			Line 716: The biosyntheses of His and Trp are not so closely related. They only have in common that they use phosphoribosediphosphate as substrate and therefore share two related enzymes.	1	2	life4041050_makarova	0
+I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			In the iron sulfur world the surface metabolism prevents the diffusion of the fixed products into the ocean. But without compartmentation soluble intermediates would escape. Furthermore, the establishment of early bioenergetics based on electrochemical Na+ gradients would have been impossible.	1	2	life4041050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			The manuscript of Eckert et al., submitted into the journal Life, concerns in principle again only one, but very important genetic criterion for evaluation of prokaryotic diversity. It should be a serious part of this complex evaluation. It is very important and useful and publication of the article is surely recommendable. However, the connection, coincidences and relations to other criteria should be better expressed.	1	2	life5010050_makarova	0
+We thus only briefly discuss names and taxonomy in our analysis.			If we accept also strains and populations from literature or from databases to the evaluation, there is always questionable the taxonomic determination (scientific names) of various species and strains, which are based on the old nomenclature and morphology. The critical revision of identification of these strains and cited materials is necessary.	1	2	life5010050_makarova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			It follows from the whole article that the authors accepted the names (and also concepts) of taxa (mostly genera) from older literature, without necessary criticism. There are included revised genera according to modern methods together with taxonomically very problematic groups. As examples is possible to mention that Aphanizomenon ovalisporum and Anabaena bergii are really very related taxa, but they were already re-classified in one special and separate genus Chrysosporum. On the other hand, the complex “Prochlorococcus/Synechococcus” is still very unclear and surely is distant from the typical genus Synechoccocus. etc.	1	2	life5010050_makarova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			Numerous cited taxa (on the generic level) were already controlled and revised by most modern methods, and examples in the manuscript should be used especially and selected from these genera. It is true, that majority of the cited genera were already revised (Microcystis, Planktothrix, Aphanizomenon), but just these taxa are important as examples and should be preferred. From the presented results follows only the confirmation of the present methodological principles and a modern system. This fact should be emphasized.	1	2	life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			It is necessary to stress the fact that the numerical criteria (e.g., exactly limited percentage similarity) never are valid uniformly for large groups of organisms and cannot be applied as one main criterion in large groups.	1	2	life5010050_makarova	0
+We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			As concerns the delimitation of taxa on the level of cyanobacterial species, there exists a lot of valuable literature, which was not cited. The characterization and separation of species was not satisfactory solved, but it is particularly important (criteria are not unique for all genera) and need better discussion.	1	2	life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			In Line 40 is cited only DeQueiroz (2007) to the problematic of species concept. It would be useful to evaluate and mention many other authors, who discussed this question more complexly. It concerns also several other problems, e.g., problematics of horizontal gene transfers (Line 43), “bounderies between taxon units” (Line 45), species concept (Line 48), etc. About species concept in cyanobacteria exist particularly many studies from last years.	1	2	life5010050_makarova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			I am not sure that cyanobacteria can be accepted as a typical example of all prokaryotes. On the contrary, they have very special position (phylogenetic, metabolic, function in nature, ultrastructural, they grow often in multicellular and differentiated thallus) and their diversity must be evaluated respecting these specificities. It is clear that the taxonomic classification must be different in different groups of organisms.	1	2	life5010050_makarova	0
+We thus only briefly discuss names and taxonomy in our analysis.			The correct citation of scientific names is the obvious request of serious scientific publications. Few unnecessary mistakes are in the manuscript, e.g., in the whole text and also in Figures is written “Arthospira” instead the correct Arthrospira, on other places must be Leptolyngbya (Table 1, Line 191), Fischerella (Line 207), Aphanizomenon (Line 218), Cylindrospermopsis (Line 219).	1	2	life5010050_makarova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			I do not understand why Leptolyngbya and Chamaesiphon could be “closely related genera”. It is in contraversion to the whole up to date results and taxonomic classification. The authors really found the “close” relation just between these two genera? By way, they both are polyphyletic and will be surely divided in several different taxa after the following precise studies.	1	2	life5010050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			It is overall well-written and innovative work, but there are few things to consider in methods and results, which should be revised, and some formal flaws. When you refer to the Synechococcus, it is important to specify, which clade you have in mind. Honda et al. (1999) and Roberston et al. (2001) showed Synechococcus is polyphyletic genus composed of a least 5 lineages. There are more recent works showing even more clades. From an amount of sequences and relationship to Prochlorococcus, I can assume, it is a marine pelagial picoplanktic clade. However, it should be specified for unexperienced reader in the field of cyanobacteria. The other polyphyletic genera should be also specified—Phormidium, Leptolyngbya, Microcoleus and others. Moreover, although the sequences might have same name (i.e., Synechococcus), they belong to polyphyletic groups, thus they probably belong to different genera, which have not yet been described.	1	2	life5010050_makarova	0
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			They might also appear to be monophyletic, because there are missing taxa between them.	1	2	life5010050_makarova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			This might also cause false positive barcoding gaps, because there would be low similarity among sequences with same name. Therefore it may largely affect results.	1	2	life5010050_makarova	0
+We thus only briefly discuss names and taxonomy in our analysis.			I suggest that the paper should be expanded with a discussion, whether barcode gaps are able to delimit described species. For example, are there two species within Cylindrospermopsis? There seems to be a barcoding gap within this clade. In this particular case, it would be beneficial to use a species epithet too to avoid confusion. Is it Cylindrospermopsis raciborskii? Does a barcoding gap in the Planktothrix clade correspond to the described species?	1	2	life5010050_makarova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			Why did you use Phormidium as an outgroup (Line 139)? Would not be more beneficial to use Gloeobacter or some other bacterium?	1	2	life5010050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			Is there any way to use barcoding gaps in a taxonomy of cyanobacteria? This might be added to discussion.	1	2	life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			Minor point: There are errors in nomenclature throughout the text, figures, and supplements, e.g. Leptolyngbya, Aphanizomenon, Arthrospira. You might check: http://www.cyanodb.cz/. It is an updated database of names of cyanobacteria.	1	2	life5010050_makarova	0
+I believe that the article with proposed changes will be useful for future research.			The manuscript of Eckert et al., submitted into the journal Life, concerns in principle again only one, but very important genetic criterion for evaluation of prokaryotic diversity. It should be a serious part of this complex evaluation. It is very important and useful and publication of the article is surely recommendable. However, the connection, coincidences and relations to other criteria should be better expressed.	1	2	life5010050_perova	0
+My critics should inform authors about more complicated problematics of this whole group, and several principles should be respected and accepted for future cyanobacterial research.			If we accept also strains and populations from literature or from databases to the evaluation, there is always questionable the taxonomic determination (scientific names) of various species and strains, which are based on the old nomenclature and morphology. The critical revision of identification of these strains and cited materials is necessary.	1	2	life5010050_perova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			It follows from the whole article that the authors accepted the names (and also concepts) of taxa (mostly genera) from older literature, without necessary criticism. There are included revised genera according to modern methods together with taxonomically very problematic groups. As examples is possible to mention that Aphanizomenon ovalisporum and Anabaena bergii are really very related taxa, but they were already re-classified in one special and separate genus Chrysosporum. On the other hand, the complex “Prochlorococcus/Synechococcus” is still very unclear and surely is distant from the typical genus Synechoccocus. etc.	1	2	life5010050_perova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			Numerous cited taxa (on the generic level) were already controlled and revised by most modern methods, and examples in the manuscript should be used especially and selected from these genera. It is true, that majority of the cited genera were already revised (Microcystis, Planktothrix, Aphanizomenon), but just these taxa are important as examples and should be preferred. From the presented results follows only the confirmation of the present methodological principles and a modern system. This fact should be emphasized.	1	2	life5010050_perova	0
+More citations of modern literature would be expectable and useful.			It is necessary to stress the fact that the numerical criteria (e.g., exactly limited percentage similarity) never are valid uniformly for large groups of organisms and cannot be applied as one main criterion in large groups.	1	2	life5010050_perova	0
+I believe that the article with proposed changes will be useful for future research.			As concerns the delimitation of taxa on the level of cyanobacterial species, there exists a lot of valuable literature, which was not cited. The characterization and separation of species was not satisfactory solved, but it is particularly important (criteria are not unique for all genera) and need better discussion.	1	2	life5010050_perova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			In Line 40 is cited only DeQueiroz (2007) to the problematic of species concept. It would be useful to evaluate and mention many other authors, who discussed this question more complexly. It concerns also several other problems, e.g., problematics of horizontal gene transfers (Line 43), “bounderies between taxon units” (Line 45), species concept (Line 48), etc. About species concept in cyanobacteria exist particularly many studies from last years.	1	2	life5010050_perova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			I am not sure that cyanobacteria can be accepted as a typical example of all prokaryotes. On the contrary, they have very special position (phylogenetic, metabolic, function in nature, ultrastructural, they grow often in multicellular and differentiated thallus) and their diversity must be evaluated respecting these specificities. It is clear that the taxonomic classification must be different in different groups of organisms.	1	2	life5010050_perova	0
+We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			The correct citation of scientific names is the obvious request of serious scientific publications. Few unnecessary mistakes are in the manuscript, e.g., in the whole text and also in Figures is written “Arthospira” instead the correct Arthrospira, on other places must be Leptolyngbya (Table 1, Line 191), Fischerella (Line 207), Aphanizomenon (Line 218), Cylindrospermopsis (Line 219).	1	2	life5010050_perova	0
+Second Round of Evaluation Round 2: I have had the more critical remarks to the manuscript; however, I think that the presented data are useful and should be published.			I do not understand why Leptolyngbya and Chamaesiphon could be “closely related genera”. It is in contraversion to the whole up to date results and taxonomic classification. The authors really found the “close” relation just between these two genera? By way, they both are polyphyletic and will be surely divided in several different taxa after the following precise studies.	1	2	life5010050_perova	0
+The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			When you refer to the Synechococcus, it is important to specify, which clade you have in mind. Honda et al. (1999) and Roberston et al. (2001) showed Synechococcus is polyphyletic genus composed of a least 5 lineages. There are more recent works showing even more clades. From an amount of sequences and relationship to Prochlorococcus, I can assume, it is a marine pelagial picoplanktic clade. However, it should be specified for unexperienced reader in the field of cyanobacteria. The other polyphyletic genera should be also specified—Phormidium, Leptolyngbya, Microcoleus and others. Moreover, although the sequences might have same name (i.e., Synechococcus), they belong to polyphyletic groups, thus they probably belong to different genera, which have not yet been described.	1	2	life5010050_perova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			They might also appear to be monophyletic, because there are missing taxa between them.	1	2	life5010050_perova	0
+The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			This might also cause false positive barcoding gaps, because there would be low similarity among sequences with same name. Therefore it may largely affect results.	1	2	life5010050_perova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			I suggest that the paper should be expanded with a discussion, whether barcode gaps are able to delimit described species. For example, are there two species within Cylindrospermopsis? There seems to be a barcoding gap within this clade. In this particular case, it would be beneficial to use a species epithet too to avoid confusion. Is it Cylindrospermopsis raciborskii? Does a barcoding gap in the Planktothrix clade correspond to the described species?	1	2	life5010050_perova	0
+Moreover, it is necessary to mention the following aspects of this manuscript: (1)			Why did you use Phormidium as an outgroup (Line 139)? Would not be more beneficial to use Gloeobacter or some other bacterium?	1	2	life5010050_perova	0
+The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			Is there any way to use barcoding gaps in a taxonomy of cyanobacteria? This might be added to discussion.	1	2	life5010050_perova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			There are errors in nomenclature throughout the text, figures, and supplements, e.g. Leptolyngbya, Aphanizomenon, Arthrospira. You might check: http://www.cyanodb.cz/. It is an updated database of names of cyanobacteria.	1	2	life5010050_perova	0
+As the authors acknowledge, the article is the first step toward a more complete understanding of an interesting class of possible chemo-physical phenomena.			Overall a beautifully written, wonderfully presented, paper with clear concepts, methods and proposals that significantly contribute to original knowledge in the field.	1	2	life5010181_perova	0
+Very well presented, clear and graphically illustrated experimental findings that make a clear case for the model.			The authors write: “In these initial stages, the emergence of bistability will serve as a proxy for the emergence of other chemical novelties in the vesicle reactor model.” The authors probably means that the existence of more than one stable regime could consent regime changes, but this situation do not indicates how these changes appear: in the article, the change is stimulated by means of a deliberated injection of chemical substances from outside. So, I think that the appearance of novelties requires a different order of considerations, whereas the bistability plays possibly the role of novelties amplification and consolidation (in particular conditions). The same observation can be done for the word “innovation”, used on the abstract and at Line 45. In order to avoid misunderstandings, I think that the authors should avoid these expressions, ad use the words “regime change”, or similar concepts.	1	2	life5010181_perova	0
+Produces a dynamic model for chemical evolution rather than traditional deterministic, static, systems that requires centralised information to direct evolution, such as a constant gradient.			Note 4 presents a very important issue: why the inner chemical environment is different from the external one, if the running chemical reactions are the same? The presence of particular features of the vesicle interior or of entrapped catalysts seem a too easy and ad hoc hypothesis. A useful reference proposing an explanation of this symmetry breaking (without requiring different conditions in internal and external environments) could be “R. Serra, M. Villani Mechanism for the formation of density gradients through semipermeable membranes Physical Review E 87, 2013”.	1	2	life5010181_perova	0
+Very well presented, clear and graphically illustrated experimental findings that make a clear case for the model.			How the authors derive the range [0.9, 101/3] for the so called “reduced surface” index? In Figure 1d, Lines 21/3 and 41/3 have some particular meanings, or are plotted mainly to ease the figure comprehension? Could the authors add some indications about the derivation of these upper limits?	1	2	life5010181_perova	0
+Produces a dynamic model for chemical evolution rather than traditional deterministic, static, systems that requires centralised information to direct evolution, such as a constant gradient.			The authors write (case 1): “of the 5000 parameter set tested under constant surface area, 82% (4098) gave a single fixed point, 321 15.7% (785) gave two fixed points, and 2.3% (117) no fixed points.” How this statistics compare with a similar one, performed on a similar chemical situation without container? Could the authors add the results of the same within a not confined environment?	1	2	life5010181_perova	0
+The mathematical methods used to deal with the problem are based on approximations and require the support of a huge number of simulations; on the other side, the approximations themselves constitute an interesting feature of the article and allow interesting discussions on the involved physical situations.			Adamala, K.; Szostak, J. ompetition betweenmodel protocells driven by an encapsulated catalyst. Nat. Chem. 2013, 5, 495–501. → “ompetition” should be: “Competition”.	1	2	life5010181_perova	0
+Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			The data are interesting and worth publishing but… (1) I don’t understand how they calculated the protein molar ratios. In the methodology section we learn that they extracted the crude proteins and then performed western analyses using specific antibodies, followed by image analyses. I don’t understand how they converted the western data to read the amount of specific proteins, in fmol, in the various organisms as shown in Figure 2; and used for the calculations in the rest of the paper. (Why are the units provided again in Line 220?). Please provide the reader with the methodology used for the quantification.	1	2	life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			PSII activity is derived from fluorescence measurement, as ETR. I am missing calibration with real measurements of PSII activity such as O2 evolution. You have got to show it for each organism you are examining otherwise it is worthless. There are many reports in the literature showing dramatic decline in fluorescence but hardly any change in O2 evolution.	1	2	life5010403_makarova	0
+This is done for a number of globally significance marine microbes of Syn and Pro lineage.			The cells were grown under a relatively low light intensity and thus the data presented here only apply to these conditions. Please make sure the reader is aware of it.	1	2	life5010403_makarova	0
+This is done for a number of globally significance marine microbes of Syn and Pro lineage.			Line 19 – I am not sure why stating the cyanobacteria are nondiazatrophic is relevant?	1	2	life5010403_makarova	0
+The data presented is robust and worthy of publication but should be discussed more critically with regard to published literature (outlined below) I suggest some minor suggestions that should be incorporated before publications.			Line 46 – (and throughout) be clear if you mean numerically dominate or dominate production – they are not always the same thing.	1	2	life5010403_makarova	0
+provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			Introduction paragraph 2 – I am unsure we know that physbobilisomes require more resource than pcb proteins – this depends on the ratio of antenna to reaction centre? Please include references if R3 this has been shown. Also, not all Pcb proteins are constitutively expressed (e.g., see Bibby et al. Nature 2003).	1	2	life5010403_makarova	0
+Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			Line 250 – This is estimated number of RUBISCO active sites – or is there evidence all the RUBSCIO you quantify is active?	1	2	life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			Figure 4 – The crunch of this paper is Figure 4e which show the Rubisco:PSII ratio is correlated to ETR. While I’m convinced in the presented data only three data-points are shown – I’m most convinced in that this same relationship is presented in in “Aquatic Photosynthesis (edition 2) Figure 7.9 – Falkowski” based on work of Sukenik (1986?). A greater discussion should be given in the text to how these datasets compare and what is significantly new in this dataset? References 35 and 36 are discussed in relation to the potential control of cytb6f on limiting electron transfer from PSII but more should be made of Falkowski’s observations which support this data.	1	2	life5010403_makarova	0
+The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			Line 308 – The data presented is used to support potential cyclic electron flow around PSI – however other alternative terminal electron sinks could also be up-regulated – these concepts are discussed in a recent review by Milligan and Behrenfeld annual review 2013 – the implications of this should be discussed unless the authors can show specifically enhanced flow around PSI.	1	2	life5010403_makarova	0
+This is done for a number of globally significance marine microbes of Syn and Pro lineage.			“In line 178 onward we read about the calibration curve. Please presnt it in the paper. Once you apply stress such as iron starvation you should recalibrate.” R4 Response:	3	2	life5010403_makarova	0
+Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			“Please present the calibration curves for the protein levels.” Response:	3	2	life5010403_makarova	0
+(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			In Figure 3, formate formula should be better symbolized as HCOO- instead of COOH- as indicated.	1	2	life5010432_makarova	0
+R2 This article will be of interest to general microbiologists, specialists in cyanobacteria, and especially researchers that conduct environmental research and also genome annotations.			Lines 59–60: Instead of links, a figure (may be included in supplementary document) will be more helpful. The links lead to a non-English website, and also they may become outdated in year or two.	1	2	life5010432_makarova	0
+R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			Line 60: Link to NO2− concentrations shows temperature profiles.	1	2	life5010432_makarova	0
+(3) p. 5 line 5, “abbreviation” has been changed to “abbreviations”.			Line 118: Was the analysis done on all FNT-proteins available at NCBI database? Please clarify.	1	2	life5010432_makarova	0
+(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			Line 40: Line 120: From the tree as it is, one can see that NitM from β-cyanobacteria form a distinct clade. However, NitM from α‐cyanobacteria are not grouped in one clade. It is possible that another outgroup sequence or different methods for alignment and tree construction will help. Line 122: What methods were used for creating alignment and phylogenetic tree (for example, neighbor-joining)?	1	2	life5010432_makarova	0
+R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			Was formate inhibition test also done on strains with nitM from CC9311, CC9605 and MIT9313?	1	2	life5010432_makarova	0
+R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			It would be especially exciting to see the uptake of nitrite at environmentally relevant concentrations (less than 50 nM)? Considering the genetic constructs available, I was wondering if the authors have attempted such experiments and what were the results.	1	2	life5010432_makarova	0
+It was particularly interesting to see the differences in protein functionality among the strains of marine Synechococcus.			In Figure 3, formate formula should be better symbolized as HCOO- instead of COOH- as indicated.	1	2	life5010432_perova	0
+The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			Lines 59–60: Instead of links, a figure (may be included in supplementary document) will be more helpful. The links lead to a non-English website, and also they may become outdated in year or two.	1	2	life5010432_perova	0
+(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			Line 60: Link to NO2− concentrations shows temperature profiles.	1	2	life5010432_perova	0
+CC9605 is an open ocean strain and does not encounter high nitrite concentrations, while CC9311 is a coastal strain that deals with a range of nitrate/nitrite availability.			Line 118: Was the analysis done on all FNT-proteins available at NCBI database? Please clarify.	1	2	life5010432_perova	0
+The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			Line 40: Line 120: From the tree as it is, one can see that NitM from β-cyanobacteria form a distinct clade. However, NitM from α‐cyanobacteria are not grouped in one clade. It is possible that another outgroup sequence or different methods for alignment and tree construction will help. Line 122: What methods were used for creating alignment and phylogenetic tree (for example, neighbor-joining)?	1	2	life5010432_perova	0
+(3) p. 5 line 5, “abbreviation” has been changed to “abbreviations”.			Was formate inhibition test also done on strains with nitM from CC9311, CC9605 and MIT9313?	1	2	life5010432_perova	0
+The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			It would be especially exciting to see the uptake of nitrite at environmentally relevant concentrations (less than 50 nM)? Considering the genetic constructs available, I was wondering if the authors have attempted such experiments and what were the results.	1	2	life5010432_perova	0
+In addition, monophyly is a guiding principle in comparing branching order with taxonomy.			I think the alignment-free methodology is interesting, however I would like to see a comparison with at least one regular alignment/treeing method, based on the same genomes the authors used, and not just a visual topological comparison with other published trees.	1	2	life5010949_makarova	0
+Authors present a new interactive tree-visualization tool which enables direct validation of taxonomic groups according to their monophyly.			As far as I can see, statistical support on the branches is missing, so I have no way of assessing if this branching order is valid. Bootstrapping or jackknifing are by no means the final word on the significance of branches, however an explanation as to how the user should assess the significance of braches would be good, i.e., branch length.	1	2	life5010949_makarova	0
+I recommend acceptance for publication in Life Sciences journal.			The fact that Thermofilum is placed outside Crenarchaeota in Figures 3 and 4 is a little disturbing. I haven’t come across such a placement in other phylogenetic analyses of Archaea, for example in Brochier-Armanet et al 2008 Nat Rev Microb, or Rinke et al. 2013 Nature. I believe this needs to better explained in the manuscript, rather than just saying “this fact is noted”.	1	2	life5010949_makarova	0
+This kind of research should be encouraged further because old taxonomic paradigms must be systematically reviewed based on new genomic data.			I am also not very convinced with the placement of Nanoarchaeota. It seems like this phylum is moving around with the addition of new sequence data (for example in Rinke et al. Figure 2 tree, they are on an entirely different branch than Euryarchaeota). Though the authors also rightfully point out that the reduced genome size may have something to do with this placement.	1	2	life5010949_makarova	0
+Round 1: and Author Response Summary Zuo et al.			The placement of Halobacteria (due to interfering Nanoarchaeota, I presume) is also a little disturbing. I would recommend that the authors provide a discussion of this. Especially with regards to other archaeal trees. For instance, in the tree of Armanet et al 2011 that the authors also refer to, the placement of Halobacteria with respect to Nanoarchaeota is very different.	1	2	life5010949_makarova	0
+Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			In summary, I find the piece interesting, but parts of the discussion are rather weak, therefore I am suggesting a major revision. Another reason for major revision is the style that the manuscript is written. I am not a native speaker, but given that I had to read sentences several times, I suspect the manuscript can benefit from an English language check.	1	2	life5010949_makarova	0
+The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			I found the revised version of this manuscript quite good, and I thank the authors for responding thoroughly to all my comments.	3	2	life5010949_makarova	0
+The tree was compared with several 16S rRNA trees reported in the literature, and the differences were minor.			I would really appreciate a “Methods” part where the CVTree is explained shortly, and the tree-viewer is explained with more detail. The absence of branch lengths and bootstraps should be discussed here. Other technical aspects of the paper (e.g., sequence dataset), parameters, criteria ... all could be well organized in this part.	1	2	life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			Archaeal phylogeny has already been studied in detail, with 16S and other marker genes, and with genomic approaches too. Some of the undersigning authors had already published on this before, although with smaller input datasets. Therefore, the fact that 16S topology is quite stable and comparable with other approaches is already known.	1	2	life5010949_makarova	0
+Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			Taxonomists have traditionally circumscribed the high taxa (specially orders and classes) with great subjectivity, i.e., without well accepted criteria. In terms of phylogenetic trees one premise has always been clear, a taxon must be monophyletic. This principle has been used in the present work to reconsider the status of some high taxa. However, authors do not explain objective criteria to properly interpret the rank of the clades, which impedes making a profound evaluation of the archaeal classification. Therefore, although authors have strong tools and dataset, they just achieved a small revision of the high taxa which is, indeed, quite biased by the underlying 16S guidelines.	1	2	life5010949_makarova	0
+Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			I have noted some lack of scientific rigor according to: many wrong taxonomic names and typos, scarce figure legends, few comments about the missing branch lengths or bootstraps (! ), redundancy in text and figures and fragments which are really difficult to understand. Authors should pay attention to language, explanations and text organization.	1	2	life5010949_makarova	0
+Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			Authors shouldn't forget (particularly in conclusion) that the resolution power of 16S for high ranks (genus and above) is currently well accepted. And the number of non-redundant 16S entries available is much much larger than that of archaeal genomes. 16s data offer a much comprehensive view of the archaeal diversity, including deep branches.	1	2	life5010949_makarova	0
+About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			- L40 “Since at present (…) are not covered (...)” is a weak reason for choosing high ranks. I recommend to shortly summarize why high ranks are so important, and why do you choose order as the lowest considered rank.	1	2	life5010949_makarova	0
+Authors present a new interactive tree-visualization tool which enables direct validation of taxonomic groups according to their monophyly.			Part 3.1 The figure legends require more rigorous explanation. Would be interesting to remember that the branch lengths are not taken into account, or were omitted.	1	2	life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			In Line 80—authors write some explanations to understand the tree figures. I would suggest to put this text before the first tree figure.	1	2	life5010949_makarova	0
+General Comments The topic is relevant for microbial taxonomy.			Figure 1 is a bit redundant. A short comment about the inclusion of that particular sequence into the Thermoplasmatales can be added into the legend of Figure 2.	1	2	life5010949_makarova	0
+Results also provide additional support to recently proposed archaeal phyla and halobacterial orders.			However, the reclassification of Methanomassiliicoccus into Thermoplasmataceae needs more explanation. To be objective, authors should address the following question, why in the same family and not in another new family?	1	2	life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			L106–107: maybe a bit inappropriate on that position. I would suggest to add a comment on the figure legend instead.	1	2	life5010949_makarova	0
+However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			Part 3.2 L109–118: Summarize and move to introduction. Those sentences are of general importance for the topic and not specific to part 3.2.	1	2	life5010949_makarova	0
+are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			L119–123: if the K issue is relevant to understand the text then please add a proper explanation. If not, then keep it simple and avoid entering into the K issue (L122–123, L132, L135, L139, 183, and also remove this K = 6 from Figure 3.)	1	2	life5010949_makarova	0
+Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			Add more explanations in legend of Figure3.	1	2	life5010949_makarova	0
+In addition, monophyly is a guiding principle in comparing branching order with taxonomy.			L143: Sounds clearer if you avoid mixing class and phylum, for example: “The placement of phylum Korarchaeota, as a closest neighbor of family Thermofilaceae, violates the monophyly of phylum Crenarchaeota.” Response:	1	2	life5010949_makarova	0
+Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			L146: No need to explain the 6+2 if it is properly explained in the figure legend. Figure 3 is redundant. I would recommend to avoid presenting different versions of the same tree; just the final tree is OK (use final/valid labels) and all important explanations in the text or legend. Perhaps the whole reasoning in Lines 142–180 is not so relevant for the current objective of comparing CVT, LTP, Bergeys? Or, perhaps, define this objective more clearly.	1	2	life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			L156–166. If I understood right, there was good support for Candidatus Aciduliprofundum as part of a clade called DHEV2, which is a sister clade of Thermoplasmatales. However in the present work the authors intend here to reclassify Aciduliprofundum into family Thermococcaceae of Thermococcales. This needs further explanation. Since the new affiliation is quite in disagreement with previous observations, and this is not properly justified in the results/discussion, the final statement “this modification would hold as long as no new facts challenge it” seems unacceptable. In addition, why should Aciduliprofundum be regarded as member of Thermococcaceae and not as another distinct family?	1	2	life5010949_makarova	0
+Even if defined in the future, it must be lineage-dependent.			L163: If I’m right the current observation actually does not support the previous work done by Brochier-Armanet.	1	2	life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			L167–173: The names are wrongly written (please check the original submission). Authors have to explain with more clarity, why is this clade of rank class. If that is the case, is it a single-order class? A single family order?	1	2	life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			L174–180: I don’t understand the reasoning along this paragraph. In addition “haolphiic_archaeon_DL31” is not well written, please be careful when copying names from other source. Also, a similar question about the objectiveness for detecting high taxa: why not to create new family?	1	2	life5010949_makarova	0
+are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			Figure 4: the reclassifications must be clearly justified in the text.	1	2	life5010949_makarova	0
+Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			L191: organisms can't be validly published, but their names.	1	2	life5010949_makarova	0
+Authors present a new interactive tree-visualization tool which enables direct validation of taxonomic groups according to their monophyly.			Part 3.3 L221–222: Sure, but authors do not provide explanations about how do they know that a clade in a tree is a family, an order, a class, etc. There is a lack of criteria to reclassify the leaves.	1	2	life5010949_makarova	0
+The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system.			L224: I don’t understand “3063 identical nucleotide positions”. Why identical?	1	2	life5010949_makarova	0
+The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system.			L239–244: hard to read, please rephrase.	1	2	life5010949_makarova	0
+In addition, monophyly is a guiding principle in comparing branching order with taxonomy.			Conclusion I disagree that CVTree approach is independent of 16S, because authors are using the current accepted classification (which is mainly 16S-based) to validate the observed clades.	1	2	life5010949_makarova	0
+The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			Why at higher ranks, genomic approaches are more effective? That needs more explanation. And authors should also consider the large benefits of 16S data availability, specially at high ranks (genus and above) where the 16S has good resolution.	1	2	life5010949_makarova	0
+present a comparative analysis of the taxonomic classification of the Archaea domain.			L208: erenow → herenow	1	2	life5010949_makarova	0
+— Done Second Round of Evaluation Round 2: and Author Response			L35-38: I don’t get well the sentences which start from “This is in contrast ... ”. Please can you specify a bit more?	3	2	life5010949_makarova	0
+In addition, monophyly is a guiding principle in comparing branching order with taxonomy.			L45: should provide → provides	3	2	life5010949_makarova	0
+General Comments The topic is relevant for microbial taxonomy.			L47–49: move to conclusions?	3	2	life5010949_makarova	0
+Even if defined in the future, it must be lineage-dependent.			L81–85: I think this paragraph is interrupting a bit the text flow. I suggest deletion.	3	2	life5010949_makarova	0
+Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			L244–245: I think the text within {} deviates the attention. I suggest delete that part, ending sentence with “monophyletic Thaumarchaeota” is also ok.	3	2	life5010949_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			"It is an interesting topic, however, the presentation seems not to be clear. It would be better to have a short but clear description about what ""bending elasticity modulus"" is and its relevance to biological functions."	1	2	life5021101_makarova	0
+According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			The theory section could be in the Method section with both principle and procedures to get kc values.	1	2	life5021101_makarova	0
+I also have questions and comments for acceptance.			However, not all results seem to be presented. For example, it is not clear what kind of images were recorded, which were used to calculate the kc values, what the sizes/shapes of the fluctuating vesicles looked like. How these observations would be different or similar to other types of vesicles.	1	2	life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			The first sentence in the introduction section should be corrected as cell should not be considered to be “the building block of life”. It’s better to re-write the “basic motivation” for the study.	1	2	life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			There are two reference lists, one for 1–31 and another from 1–34. There are repeated ones. It’s not clear why the case was.	1	2	life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.	3	2	life5021101_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			If only measured at one temperature, how would it be possible to draw a conclusion with “thermally induced shape fluctuations”?	3	2	life5021101_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			Is it possible to do the measurement at elevated temperatures?	3	2	life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			Would kc be temperature dependent? If yes, what would be the valid range?	3	2	life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			The photo added merely shows the shape, but is it possible to show a few time related changes of the shape at different temperatures?	3	2	life5021101_makarova	0
+The observed fluctuations of the membrane do not destabilize it.			The numbering from Lines 306–307 may not be correct as there are two 2.2.	3	2	life5021101_makarova	0
+I also have questions and comments for acceptance.			In addition, the number of tables and figures are too low for article. If could, the author should add other experimental data.	1	2	life5021101_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			In Page 1, Lines 27–28, the author described “the experimental set-up was improved …”. However, the authors have already reported the measurements of bending elasticity using stroboscopic illumination in author’s previous papers (such as Genova et al., J. Optoelectro. Adv. Mat., 2005). Describe how the experimental set-up was improved in this paper more precisely.	1	2	life5021101_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			As a comparison of archaeal lipids, SOPC was used in this study. SOPC contains an unsaturated bond in its alkyl chains and its transition temperature seems to be quite low compared with archaeal lipids. What is the reason why the authors selected the SOPC for the representative of eukaryotic lipids?	1	2	life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			In page 4, between line 108 and 109, this equation should be numbered as (6).	1	2	life5021101_makarova	0
+I also have questions and comments for acceptance.			Describe the temperature condition for measurement of fluctuation of GVs. Are there any effects of temperature on the fluctuation of GVs?	1	2	life5021101_makarova	0
+The observed fluctuations of the membrane do not destabilize it.			In this paper, bending elasticity modulus was measured in pure water. Are there any effects of ionic strength or composition of solution on the fluctuation of GVs?	1	2	life5021101_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			The manuscript was thoroughly revised according to reviewer’s suggestions and comments. Now this revised manuscript can be acceptable for publication.	3	2	life5021101_makarova	0
+The observed fluctuations of the membrane do not destabilize it.			"It would be better to have a short but clear description about what ""bending elasticity modulus"" is and its relevance to biological functions."	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			The theory section could be in the Method section with both principle and procedures to get kc values.	1	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			However, not all results seem to be presented. For example, it is not clear what kind of images were recorded, which were used to calculate the kc values, what the sizes/shapes of the fluctuating vesicles looked like. How these observations would be different or similar to other types of vesicles.	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			We have rewritten the introductory paragraph (Lines 46–60).	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			There are two reference lists, one for 1–31 and another from 1–34. There are repeated ones. It’s not clear why the case was.	1	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones. The observed fluctuations of the membrane do not destabilize it. However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.	3	2	life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			If only measured at one temperature, how would it be possible to draw a conclusion with “thermally induced shape fluctuations”?	3	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			Is it possible to do the measurement at elevated temperatures?	3	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			Would kc be temperature dependent? If yes, what would be the valid range?	3	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			The photo added merely shows the shape, but is it possible to show a few time related changes of the shape at different temperatures?	3	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			The numbering from Lines 306–307 may not be correct as there are two 2.2.	3	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids. Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs. According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs. The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers. In addition, the number of tables and figures are too low for article. If could, the author should add other experimental data.	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			I also have questions and comments for acceptance. The list of comments is as follows. In Page 1, Lines 27–28, the author described “the experimental set-up was improved …”. However, the authors have already reported the measurements of bending elasticity using stroboscopic illumination in author’s previous papers (such as Genova et al., J. Optoelectro. Adv. Mat., 2005). Describe how the experimental set-up was improved in this paper more precisely.	1	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			As a comparison of archaeal lipids, SOPC was used in this study. SOPC contains an unsaturated bond in its alkyl chains and its transition temperature seems to be quite low compared with archaeal lipids. What is the reason why the authors selected the SOPC for the representative of eukaryotic lipids?	1	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			In page 4, between line 108 and 109, this equation should be numbered as (6).	1	2	life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			Describe the temperature condition for measurement of fluctuation of GVs. Are there any effects of temperature on the fluctuation of GVs?	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			In this paper, bending elasticity modulus was measured in pure water. Are there any effects of ionic strength or composition of solution on the fluctuation of GVs?	1	2	life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			The manuscript was thoroughly revised according to reviewer’s suggestions and comments. Now this revised manuscript can be acceptable for publication.	3	2	life5021101_perova	0
+This is a great paper to aim at such a direction.			However, being experts in their field the authors might not be aware that for readers less familiar with the metabolism/physiology of archaea, the examples are not always easy to follow, such as examples given on p. 7 starting line 196 (The main difference here is between a ferredoxin-dependent and/or a NAD-dependent decarboxylation of pyruvate into acetyl-CoA?/A few lines later when I read that “halophilic archaea do not contain the coenzyme methanopterin”, I had to go back and reread the preceding sentences to get an idea of the argument. ), p. 10 starting Line 299, and p. 12 starting Line 354.	1	2	life5021427_makarova	0
+R2 Round 1: and Author Response I received this manuscript and looked at it with joy.			There is now available a great resource for the gene discovery in Haloferax volcanii (Kiljunen et al 2014: BMC Biology). A comprehensive insertion mutant library has been generated for this model archaeon in order to facilitate gene discovery. This should be mentitioned and discussed in the text; i.e., how this resource could be used to assign functions for genes.	1	2	life5021427_makarova	0
+The authors are to be complimented for making the effort to provide well curated annotations for a set of haloarchaeal genomes, and contribute to improving public databases (EMBL/Genbank, UniProt).			Giving specific examples is certainly a good idea. However, being experts in their field the authors might not be aware that for readers less familiar with the metabolism/physiology of archaea, the examples are not always easy to follow, such as examples given on p. 7 starting line 196 (The main difference here is between a ferredoxin-dependent and/or a NAD-dependent decarboxylation of pyruvate into acetyl-CoA?/A few lines later when I read that “halophilic archaea do not contain the coenzyme methanopterin”, I had to go back and reread the preceding sentences to get an idea of the argument. ), p. 10 starting Line 299, and p. 12 starting Line 354.	1	2	life5021427_perova	0
+Importance: Archaea is/are the third domain of life, no more or less!			There is now available a great resource for the gene discovery in Haloferax volcanii (Kiljunen et al 2014: BMC Biology). A comprehensive insertion mutant library has been generated for this model archaeon in order to facilitate gene discovery. This should be mentitioned and discussed in the text; i.e., how this resource could be used to assign functions for genes.	1	2	life5021427_perova	0
+The other potentially inactivating components of the plasma are not specified in the manuscript.			The plasma conditions are not described sufficiently. It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are. The term amount is not applicable here. With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.	1	2	life6020022_makarova	0
+Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Line 57: Are you sure that the Martian Analog soils were sterile before using?			The other potentially inactivating components of the plasma are not specified in the manuscript. For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description. Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.	1	2	life6020022_makarova	0
+In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.			Three different Mars regolith simulants were used. The mineralogical composition is given in table, but not the grain size distribution. Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.	1	2	life6020022_makarova	0
+The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).			The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.	1	2	life6020022_makarova	0
+The other potentially inactivating components of the plasma are not specified in the manuscript.			First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_makarova	0
+For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description.			The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).	1	2	life6020022_makarova	0
+While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.			In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.	1	2	life6020022_makarova	0
+The exposure times varied from 1.5 to 45 min with different experiment Conditions/procedures without resulting in a significant inactivating effect on the bacterial cells.			Lines 31-32: Bacillus subtilis and B.pumilus should be written in italic.	1	2	life6020022_makarova	0
+While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.			Line 38: Deinococcus radiodurans should be written in italic. While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.	1	2	life6020022_makarova	0
+Author Response Reviwers comments The manuscript contains scientific material that can be published as a short communication.			Line 41: Medium instead of media. If I understood correctly, you used only one medium.	1	2	life6020022_makarova	0
+With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.			Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Reply:	1	2	life6020022_makarova	0
+With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.			Line 57: Are you sure that the Martian Analog soils were sterile before using?	1	2	life6020022_makarova	0
+In the manuscript ‘Testing the Effects of Low-Temperature Plasma-Sterilization on Mars Analog Soil Samples Intermixed with Deinococcus radiodurans’ by Schirmack et al., the results of the application of low-temperature plasma on Mars regolth simulant mixed with cells of the bacterium D. radiodurans were reported.			Line 78: Please explain in the text what is “a vacuum treatment control”.	1	2	life6020022_makarova	0
+However, there are some remarks on the justification of objectives of the study and presentation of the results.			Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?	1	2	life6020022_makarova	0
+Minor comments (please see changes in text at the relevant lines)			The plasma conditions are not described sufficiently. It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are. The term amount is not applicable here. With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.	1	2	life6020022_perova	0
+However, there are some remarks on the justification of objectives of the study and presentation of the results.			The other potentially inactivating components of the plasma are not specified in the manuscript. For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description. Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.	1	2	life6020022_perova	0
+Three different Mars regolith simulants were used.			Three different Mars regolith simulants were used. The mineralogical composition is given in table, but not the grain size distribution. Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.	1	2	life6020022_perova	0
+The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_perova	0
+Two different gas mixtures were used to produce plasma, argon-nitrogen and argon-oxygen.			First of all it is necessary to change the title and at least remove “The testing”.	1	2	life6020022_perova	0
+As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV.			The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).	1	2	life6020022_perova	0
+In the manuscript ‘Testing the Effects of Low-Temperature Plasma-Sterilization on Mars Analog Soil Samples Intermixed with Deinococcus radiodurans’ by Schirmack et al., the results of the application of low-temperature plasma on Mars regolth simulant mixed with cells of the bacterium D. radiodurans were reported.			Line 38: Deinococcus radiodurans should be written in italic. While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.	1	2	life6020022_perova	0
+In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.			Line 41: Medium instead of media. If I understood correctly, you used only one medium.	1	2	life6020022_perova	0
+Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Line 57: Are you sure that the Martian Analog soils were sterile before using?			Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Reply:	1	2	life6020022_perova	0
+The introduction part requires more arguments to prove choice of the problem and the research object (D. radiodurans).			Line 57: Are you sure that the Martian Analog soils were sterile before using?	1	2	life6020022_perova	0
+The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			Line 78: Please explain in the text what is “a vacuum treatment control”.	1	2	life6020022_perova	0
+Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.			How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?	1	2	life6020022_perova	0
+Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included.			line 38 - The linking of 'other molecules' to 'phenotype' here is a little odd. There may be a link from one gene to one protein, but one protein is not really a phenotype. There are many quantitative traits affected by many genes.	1	2	life6030029_makarova	0
+According to the abstract, this manuscript covers a number of different areas relating to the RNA world hypothesis, including its compatibility with (likely) primitive Earth conditions; the likely characteristics of an RNA world; and, finally, the relationship between an early life-like RNA world system(s) and other life-like systems.			line 73 - the point is made that current RNA World theories do not seem compatible with a hydrothermal origin of life. But it is not clear where this leads us. Maybe life did not begin with RNA. Maybe life did not begin with hydrothermal vents. Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable. Maybe there is some slightly different nucleic acid-like polymer that is stable in hydrothermal conditions. Too many unknowns here to be a useful point.	1	2	life6030029_makarova	0
+Section 3.4 - 'There is a debate whether CMIO could have formed by a CCSI consisting entirely or mainly of RNA'.			lines 93-106 - I mostly agree with this paragraph but it is not said very clearly. Certainly we need to distinguish between long peptides formed spontaneously and those that are translated. But several questions still remain - were there long peptides before long RNA strands? Was there a way of reproducibly making the same amino acid sequence without translating it from RNA? If not, then is there any way that random non-encoded peptides could be useful to RNAs?	1	2	life6030029_makarova	0
+Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable.			The idea in caption to Fig 4 that building blocks do not interact directly with the environment seems interesting and plausible, but not very well defined. Are you saying a cell interacts with the environment but a single gene does not? Or a multicellular organism interacts but one cell does not? The citations 52 and 53 are in fields that are not related to biology and origin of life. So I think this idea needs to be explained and justified in a biological context.	1	2	life6030029_makarova	0
+Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable.			Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point. The abstract promised to talk about RNA World and the origin of life. At this point the article seems to have strayed a long way from this intention. From the origin of life viewpoint, even the simplest of these (the prokaryote) is still very complex. The issue for the origin of life is how to get to a prokaryote. I don't really think that looking at social insects and human societies will help much in understanding the origin of life. The ideas of CCSI and CMIO seem interesting, but they are discussed with the high-level examples (pages 10-11), not with molecular and cellular examples. Probably a lot of this could be simplified. It is only when we get to paragraph 3.1 that we get to the point of the article.	1	2	life6030029_makarova	0
+chemistry background (and has published a number of physical chemistry studies investigating the compatibility of the RNA world hypothesis with a hydrothermal vent origin of life), but has also published extensively in the social sciences area, for example applying evolutionary concepts to other 'life-like' systems, such as social insect colonies and human civilizations.			Fig 7 - Viewing a metabolism as a cycle seems to be only half the story. Nutrients go in and waste comes out. This is a throughput, not a cycle. Also the diagram does not indicate whether the ribozymes are made by the cycle. I think there is some room for improvement in this diagram.	1	2	life6030029_makarova	0
+The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol.			Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.	1	2	life6030029_makarova	0
+In summary, I think there are a lot of interesting points here, but the article could use some streamlining to emphasize the key new ideas.			Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included. For example there must be ways of making nucleotides from scratch. It cannot be true that the only source of nucleotides is by degrading oligomers.	1	2	life6030029_makarova	0
+Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.			Section 3.4 - 'There is a debate whether CMIO could have formed by a CCSI consisting entirely or mainly of RNA'. This section seems to mix up two important questions. (i) Are RNA catalysts sufficient? Do we need other kinds of biomolecules like protein catalysts? (ii) Do we need the RNA system to be enclosed in a cell membrane or other kind of compartment? If so, can RNA control the growth and division of the membrane? These issues need to be considered separately.	1	2	life6030029_makarova	0
+Or a multicellular organism interacts but one cell does not?			line 554 - The case that two genes were essential initially is not made. Can we not have an RNA polymerase that uses other copies of itself as a template? If chemistry supplies monomers, then only one gene is necessary. The input and output would not necessarily need to be controlled by another gene. It would seem difficult to evolve two separate ribozyme functions at the same time.	1	2	life6030029_makarova	0
+ lines 93-106 - I mostly agree with this paragraph but it is not said very clearly.			If there were two genes, what were the functions of the two genes? The connection between the reactions catalyzed by the ribozymes and the concepts of CCSI and CMIO is not very clear at this point.	1	2	life6030029_makarova	0
+Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.			The conclusion comes back to the point about primitive Earth environment. This links to the introductory section about hydrothermal conditions etc. I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question. It is only mentioned in the introduction and conclusions.	1	2	life6030029_makarova	0
+The citations 52 and 53 are in fields that are not related to biology and origin of life.			One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused. It is also fairly rambling: for example, lines 95-106 could be removed,	1	2	life6030029_makarova	0
+The ideas of CCSI and CMIO seem interesting, but they are discussed with the high-level examples (pages 10-11), not with molecular and cellular examples.			"The English is a problem too: ""soft molecules"" is not a typical way of describing organic molecules, and ""formation and deformation"" of molecules would be better expressed as formation and degradation. Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."	1	2	life6030029_makarova	0
+The citations 52 and 53 are in fields that are not related to biology and origin of life.			More fundamentally perhaps, the distinction that is made between CCSI and CMIO is, as the author himself notes, remarkably similar to that between genotype and phenotype, and information and function, and even replication and metabolism. The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol. 364:249-59). Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.	1	2	life6030029_makarova	0
+Are you saying a cell interacts with the environment but a single gene does not?			line 38 - The linking of 'other molecules' to 'phenotype' here is a little odd. There may be a link from one gene to one protein, but one protein is not really a phenotype. There are many quantitative traits affected by many genes.	1	2	life6030029_perova	0
+The issue for the origin of life is how to get to a prokaryote.			line 73 - the point is made that current RNA World theories do not seem compatible with a hydrothermal origin of life. But it is not clear where this leads us. Maybe life did not begin with RNA. Maybe life did not begin with hydrothermal vents. Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable. Maybe there is some slightly different nucleic acid-like polymer that is stable in hydrothermal conditions. Too many unknowns here to be a useful point.	1	2	life6030029_perova	0
+It is only mentioned in the introduction and conclusions.			lines 93-106 - I mostly agree with this paragraph but it is not said very clearly. Certainly we need to distinguish between long peptides formed spontaneously and those that are translated. But several questions still remain - were there long peptides before long RNA strands? Was there a way of reproducibly making the same amino acid sequence without translating it from RNA? If not, then is there any way that random non-encoded peptides could be useful to RNAs?	1	2	life6030029_perova	0
+It is only when we get to paragraph 3.1 that we get to the point of the article.			The idea in caption to Fig 4 that building blocks do not interact directly with the environment seems interesting and plausible, but not very well defined. Are you saying a cell interacts with the environment but a single gene does not? Or a multicellular organism interacts but one cell does not? The citations 52 and 53 are in fields that are not related to biology and origin of life. So I think this idea needs to be explained and justified in a biological context.	1	2	life6030029_perova	0
+Do we need other kinds of biomolecules like protein catalysts?			Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point. The abstract promised to talk about RNA World and the origin of life. At this point the article seems to have strayed a long way from this intention. From the origin of life viewpoint, even the simplest of these (the prokaryote) is still very complex. The issue for the origin of life is how to get to a prokaryote. I don't really think that looking at social insects and human societies will help much in understanding the origin of life. The ideas of CCSI and CMIO seem interesting, but they are discussed with the high-level examples (pages 10-11), not with molecular and cellular examples. Probably a lot of this could be simplified. It is only when we get to paragraph 3.1 that we get to the point of the article.	1	2	life6030029_perova	0
+It is only mentioned in the introduction and conclusions.			Fig 7 - Viewing a metabolism as a cycle seems to be only half the story. Nutrients go in and waste comes out. This is a throughput, not a cycle. Also the diagram does not indicate whether the ribozymes are made by the cycle. I think there is some room for improvement in this diagram.	1	2	life6030029_perova	0
+chemistry background (and has published a number of physical chemistry studies investigating the compatibility of the RNA world hypothesis with a hydrothermal vent origin of life), but has also published extensively in the social sciences area, for example applying evolutionary concepts to other 'life-like' systems, such as social insect colonies and human civilizations.			Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.	1	2	life6030029_perova	0
+It is only when we get to paragraph 3.1 that we get to the point of the article.			Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included. For example there must be ways of making nucleotides from scratch. It cannot be true that the only source of nucleotides is by degrading oligomers.	1	2	life6030029_perova	0
+If so, can RNA control the growth and division of the membrane?			Section 3.4 - 'There is a debate whether CMIO could have formed by a CCSI consisting entirely or mainly of RNA'. This section seems to mix up two important questions. (i) Are RNA catalysts sufficient? Do we need other kinds of biomolecules like protein catalysts? (ii) Do we need the RNA system to be enclosed in a cell membrane or other kind of compartment? If so, can RNA control the growth and division of the membrane? These issues need to be considered separately.	1	2	life6030029_perova	0
+The second point is that the paper contains too many errors.			line 554 - The case that two genes were essential initially is not made. Can we not have an RNA polymerase that uses other copies of itself as a template? If chemistry supplies monomers, then only one gene is necessary. The input and output would not necessarily need to be controlled by another gene. It would seem difficult to evolve two separate ribozyme functions at the same time.	1	2	life6030029_perova	0
+Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.			If there were two genes, what were the functions of the two genes? The connection between the reactions catalyzed by the ribozymes and the concepts of CCSI and CMIO is not very clear at this point.	1	2	life6030029_perova	0
+However, I find the manuscript contains a number of weaknesses: One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused.			The conclusion comes back to the point about primitive Earth environment. This links to the introductory section about hydrothermal conditions etc. I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question. It is only mentioned in the introduction and conclusions.	1	2	life6030029_perova	0
+It is only mentioned in the introduction and conclusions.			One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused. It is also fairly rambling: for example, lines 95-106 could be removed,	1	2	life6030029_perova	0
+Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point.			"The English is a problem too: ""soft molecules"" is not a typical way of describing organic molecules, and ""formation and deformation"" of molecules would be better expressed as formation and degradation. Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."	1	2	life6030029_perova	0
+The abstract promised to talk about RNA World and the origin of life.			More fundamentally perhaps, the distinction that is made between CCSI and CMIO is, as the author himself notes, remarkably similar to that between genotype and phenotype, and information and function, and even replication and metabolism. The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol. 364:249-59). Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.	1	2	life6030029_perova	0
+It is shown that integrated MO isolators consist of Dy:CeIG thin films show better temperature stability compared to Ce:YIG based devices in the temperature range of 23 ℃ to 70 ℃.			It is not indicated in the introduction whether any alternative to thin films of yttrium iron garnet is currently being considered for use as magneto-optical resonators.	1	2	ma15051691_makarova	0
+Journal of Applied Physics 1994, 75, 6103-6105.			How will the temperature dependence of the Faraday rotation hysteresis loops of Dy:CeYIG thin films change when moving to another wavelength range? Is there any dependence on the wavelength?	1	2	ma15051691_makarova	0
+1) It is not indicated in the introduction whether any alternative to thin films of yttrium iron garnet is currently being considered for use as magneto-optical resonators. 2) How will the temperature dependence of the Faraday rotation hysteresis loops of Dy:CeYIG thin films change when moving to another wavelength range?			How can one explain the increase in the value of the Faraday rotation angle in the range of 30-40 degrees for Dy:CeYIG (Fig. 3f), which is absent for Ce:YIG (Fig. 3f)?	1	2	ma15051691_makarova	0
+It is shown that integrated MO isolators consist of Dy:CeIG thin films show better temperature stability compared to Ce:YIG based devices in the temperature range of 23 ℃ to 70 ℃.			"For several references, the text ""Error! Reference source not found"" appears. Please check with the editor if this can be fixed."	1	2	ma15051691_makarova	0
+Please check with the editor if this can be fixed.			In equation (2) and the following line, there are inconcistencies in the use of capital letters for the Faraday rotation angles.	1	2	ma15051691_makarova	0
+It is shown that integrated MO isolators consist of Dy:CeIG thin films show better temperature stability compared to Ce:YIG based devices in the temperature range of 23 ℃ to 70 ℃.			"Line 111. ""X-ray diffraction patterns"", not ""spectra""."	1	2	ma15051691_makarova	0
+5) Figures 3 (a) and (b): It could be good to use the same horizontal scale for a clearer comparison.			Figures 2 (c) and (d). While the Faraday rotation angle appears to decrease more or less linearly with temperature in the Ce:YIG film (as far as can be inferred from the non-linear horizontal scale), the decrease is not taking place at the same rate in the Dy:Ce:YIG film. What causes this different behavior?	1	2	ma15051691_makarova	0
+"The article ""Dysprosium substituted Ce:YIG thin films for temperature in-2 sensitive integrated optical isolator applications contains the results"" presents a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications."			Figures 3 (a) and (b): It could be good to use the same horizontal scale for a clearer comparison.	1	2	ma15051691_makarova	0
+I recommend publishing the manuscript after minor the revision.			How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_makarova	0
+If so, do the decomposition products form bubbles in your solvent?			The authors mention cracks as well as voids. How are the cracks differentiated from the voids? Were voids observed? Could computed microtomography be used to confirm some of the experimental results?	1	2	ma15124258_makarova	0
+Or, are both the number and size of the defect increasing.			Are your pores assumed to be spherical? Are cracks cylindrical?	1	2	ma15124258_makarova	0
+How do these experiments differentiate between increasing the number of defects and the size of the defects?			I would be interesting to have additional support for the process by using thermal experiments such as differential scanning calorimetry (DSC). How do you results compare to phase transformation measured using DSC? Are the onsets and finishing of the phase transformations consistent with the DSC measurements?	1	2	ma15124258_makarova	0
+If so, do the decomposition products form bubbles in your solvent?			Do your experiments (WAXS and SAXS) measure the defect size and number of defects or is a model used to infer these quantities?	1	2	ma15124258_makarova	0
+Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			What do the colors in your figures represent?	1	2	ma15124258_makarova	0
+Could the increase be due to larger defects rather than an increase in the number of defects?			Could there be any other explanation rather than increase in number of internal nano-scale pores? Could the increase be due to larger defects rather than an increase in the number of defects? If so, how do you prove this? Could you use X-ray microtomography on your samples?	1	2	ma15124258_makarova	0
+I also noticed that your references did not include any of the more recent work published in 2021 or 2022.			Is the temperature high enough to cause decomposition effects? If so, do the decomposition products form bubbles in your solvent? If so, could this bias your conclusions?	1	2	ma15124258_makarova	0
+Is it the number of defects that are increasing?			Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?	1	2	ma15124258_makarova	0
+I had several questions regarding this work: 1.			"Could you be more specific on the ""spherical model"" on line 170? Co you assume the pore volume remains unchanged, but the number increases? or visa verse?"	1	2	ma15124258_makarova	0
+The conclusion must be supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after temperature effects.			Again, what is happening? Are the number of pores increasing or is the existing pores increasing in volume?	1	2	ma15124258_makarova	0
+As you can tell by these questions, my main issue is related to the specific form of the thermal damage.			I also noticed that your references did not include any of the more recent work published in 2021 or 2022. There are some more recent articles that would be worth citing.	1	2	ma15124258_makarova	0
+The authors present an experimental study of nano-scale defects in CL-20 under heating.			In the introduction, the authors foreground their work, focusing on the relationship between the presence of pores and sensitivity. It indeed the case, but I could not find any discussion on this issue. Therefore, I suggest include some comments on this and, probably, provide some discussion of how to avoid the increase of such defects, which increase sensitivity of energetic crystals, in particular CL-20.	1	2	ma15124258_makarova	0
+Every available suggestion has been answered carefully and corrected in the article.			Section 2.1. Materials and instruments, you need to specify the particle size of the initial CL-20 powders.	1	2	ma15124258_makarova	0
+Is the temperature high enough to cause decomposition effects?			“The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.	1	2	ma15124258_makarova	0
+Response to Reviewer' Comments Dear Reviewer, We sincerely appreciate your comments and suggestions on the manuscript!			Page 6. 199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.	1	2	ma15124258_makarova	0
+Is it the number of defects that are increasing?			The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.	1	2	ma15124258_makarova	0
+The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_perova	0
+199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.”From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.			The authors mention cracks as well as voids. How are the cracks differentiated from the voids? Were voids observed? Could computed microtomography be used to confirm some of the experimental results?	1	2	ma15124258_perova	0
+The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			Are your pores assumed to be spherical? Are cracks cylindrical. Are they statistical in nature?	1	2	ma15124258_perova	0
+Co you assume the pore volume remains unchanged, but the number increases?			I would be interesting to have additional support for the process by using thermal experiments such as differential scanning calorimetry (DSC). How do you results compare to phase transformation measured using DSC? Are the onsets and finishing of the phase transformations consistent with the DSC measurements?	1	2	ma15124258_perova	0
+Thus, I can recommend it for publication after a minor revision.			5. Do your experiments (WAXS and SAXS) measure the defect size and number of defects or is a model used to infer these quantities?	1	2	ma15124258_perova	0
+Could the increase be due to larger defects rather than an increase in the number of defects?			6. What do the colors in your figures represent?	1	2	ma15124258_perova	0
+“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20.			7. Could there be any other explanation rather than increase in number of internal nano-scale pores? Could the increase be due to larger defects rather than an increase in the number of defects? If so, how do you prove this? Could you use X-ray microtomography on your samples?	1	2	ma15124258_perova	0
+Or, is it the volume of the defect that is increasing?			8. Is the temperature high enough to cause decomposition effects? If so, do the decomposition products form bubbles in your solvent? If so, could this bias your conclusions?	1	2	ma15124258_perova	0
+To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”			9. Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?	1	2	ma15124258_perova	0
+Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?			"10. Could you be more specific on the ""spherical model"" on line 170? Co you assume the pore volume remains unchanged, but the number increases? or visa verse?"	1	2	ma15124258_perova	0
+Response to Reviewer' Comments Dear Reviewer, We sincerely appreciate your comments and suggestions on the manuscript!			12. Again, what is happening? Are the number of pores increasing or is the existing pores increasing in volume?	1	2	ma15124258_perova	0
+Could the increase be due to larger defects rather than an increase in the number of defects?			I also noticed that your references did not include any of the more recent work published in 2021 or 2022. There are some more recent articles that would be worth citing.	1	2	ma15124258_perova	0
+If so, do the decomposition products form bubbles in your solvent?			How do the WAXS and SAXS differ? How do these experiments differentiate between increasing the number of defects and the size of the defects?	1	2	ma15124258_perova	0
+199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.”From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.			1. Section 2.1. Materials and instruments, you need to specify the particle size of the initial CL-20 powders.	1	2	ma15124258_perova	0
+Or, are both the number and size of the defect increasing.			2. “The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.	1	2	ma15124258_perova	0
+“The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.			3. Page 6. 199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.	1	2	ma15124258_perova	0
+Every available suggestion has been answered carefully and corrected in the article.			4. The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.	1	2	ma15124258_perova	0
+Medicina 2021, 57, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Lastname Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.			First is the omission of any items referring to experience/use of eCigarette/vaping. Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.	1	2	medicina58040502_makarova	0
+Please provide the most important results in the text, and the rest of them are present in Tables.			"Second it a lack of analysis of second-hand smoke exposure. Specifically, whether exposure increases risk of smoking. Correlations do not confirm causality. It is likely that students who smoke actively seek roommates who smoke, and students who do not smoke seek roommates who do not smoke. Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""	1	2	medicina58040502_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			Finally, I found it disturbing that medical school faculty tolerate student smoking at all. This is contrary to accepted public health policy internationally. Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous. The link between smoking and reduced life expectancy has been shown to be causal. This result calls into question the university's curriculum addressing the very many diseases associated with tobacco smoking - most of which reduce life expectancy. This is the opposite of health promotion one expects from all healthcare providers - especially physicians.	1	2	medicina58040502_makarova	0
+Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status.			"The study aim should be more precise ""The aim of this study was...."""	1	2	medicina58040502_makarova	0
+This is the opposite of health promotion one expects from all healthcare providers - especially physicians.			Study sample - please provide more precise data on population (faculties at the university and its share in the total sample of recruited subjects)	1	2	medicina58040502_makarova	0
+Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.			Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status	1	2	medicina58040502_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			The results section is too extensive - please avoid describing all the results in the text. Please provide the most important results in the text, and the rest of them are present in Tables.	1	2	medicina58040502_makarova	0
+Finally, I found it disturbing that medical school faculty tolerate student smoking at all.			In table 5 please provide thr number of subjects (n=.....)	1	2	medicina58040502_makarova	0
+Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.			Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals)	1	2	medicina58040502_makarova	0
+"The study aim should be more precise ""The aim of this study was...."" Response 1."			Please provide practical implications of this study	1	2	medicina58040502_makarova	0
+This is contrary to accepted public health policy internationally.			Please provide a conclusion based on obtained findings and please avoid overwhelming conclusions	1	2	medicina58040502_makarova	0
+In Table 5 please provide thr number of subjects (n=.....) Response 5.			is the omission of any items referring to experience/use of eCigarette/vaping. Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.	1	2	medicina58040502_perova	0
+"The study aim should be more precise ""The aim of this study was...."" Response 1."			"lack of analysis of second-hand smoke exposure. Specifically, whether exposure increases risk of smoking. Correlations do not confirm causality. It is likely that students who smoke actively seek roommates who smoke, and students who do not smoke seek roommates who do not smoke. Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""	1	2	medicina58040502_perova	0
+Results were consistent with previous studies of similar target populations.			Finally, I found it disturbing that medical school faculty tolerate student smoking at all. This is contrary to accepted public health policy internationally. Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous. The link between smoking and reduced life expectancy has been shown to be causal. This result calls into question the university's curriculum addressing the very many diseases associated with tobacco smoking - most of which reduce life expectancy. This is the opposite of health promotion one expects from all healthcare providers - especially physicians.	1	2	medicina58040502_perova	0
+In Table 5 please provide thr number of subjects (n=.....) Response 5.			"The study aim should be more precise ""The aim of this study was...."""	1	2	medicina58040502_perova	0
+Specifically, whether exposure increases risk of smoking.			Study sample - please provide more precise data on population (faculties at the university and its share in the total sample of recruited subjects)	1	2	medicina58040502_perova	0
+Please provide practical implications of this study Response 7.			Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status	1	2	medicina58040502_perova	0
+Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals) Response 6.			The results section is too extensive - please avoid describing all the results in the text. Please provide the most important results in the text, and the rest of them are present in Tables.	1	2	medicina58040502_perova	0
+Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals) Response 6.			In table 5 please provide thr number of subjects (n=.....)	1	2	medicina58040502_perova	0
+"Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""			Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals)	1	2	medicina58040502_perova	0
+Please provide the most important results in the text, and the rest of them are present in Tables.			Please provide practical implications of this study	1	2	medicina58040502_perova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			Please provide a conclusion based on obtained findings and please avoid overwhelming conclusions	1	2	medicina58040502_perova	0
+They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			My main issue is with the way the results are presented. The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value. Even if they performed a test, important questions remain on the selection of the reference. The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry. The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA. My suggestion is to offer a better background for these results by trying to answer two main questions:	1	2	medsci4030012_makarova	0
+They found that the burden of deletions in these patients is much higher than expected, as compared with CNV burden in HapMap populations.			1) is the deletion burden observed really exceptional in a population sample of similar ancestry? To answer this question, I would refer to the 1000 genomes data where there are Mexicans, Colombians, Puerto Ricans and Peruvians. There must be available CNV estimations for these ancestries. If you find good estimates then perhaps a statistical tests with those as your control group would be more believable.	1	2	medsci4030012_makarova	0
+Studies on the 1000 genomes show differences between Puerto Ricans, Colombians, Mexicans and Peruvians.			2) is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies. Please cite those.	1	2	medsci4030012_makarova	0
+The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA.			The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract. I suggest bringing this result up to the surface and making it more relevant even from the abstract. You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.	1	2	medsci4030012_makarova	0
+The authors need to make clear about “pre-, cancer (91.3%) groups”.			I missed a definition of the sample's ancestry. In the title it refers to Mexican American but in the rest of the manuscript it is treated as a latino population. Note that there are differences in admixture in the latino population depending of their country of origin. Studies on the 1000 genomes show differences between Puerto Ricans, Colombians, Mexicans and Peruvians. Therefore, if the authors can state that their sample is essentially Mexican that would be more informative and precise. I would then limit the use of latino. However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.	1	2	medsci4030012_makarova	0
+They observed that their sample is enriched in CNV variants in pathways relevant to the phenotype like endometrial cancer and estrogen signaling pathway.			Abstract Line 23: should read Latino Populations	1	2	medsci4030012_makarova	0
+They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			Figure 1: I cannot see the supposed deletions marked in green	1	2	medsci4030012_makarova	0
+My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			Table 2b: Please remove the % sign in some of the table cells, for instance in Reference (Deletion, 10-100kb).	1	2	medsci4030012_makarova	0
+The authors need to make clear about “pre-, cancer (91.3%) groups”.			Line 34. The authors need to make clear about “pre-, cancer (91.3%) groups”. It is not clear if there is one or two groups but just one percentage.	1	2	medsci4030012_makarova	0
+My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			Line 40. The authors may add up to 6 key words.	1	2	medsci4030012_makarova	0
+It is not clear which statistical method is used for the significant results in Table 2.			Lines 196-197. It is not clear which statistical method is used for the significant results in Table 2.Please give more details.	1	2	medsci4030012_makarova	0
+2) is the deletion burden observed similar to those for other cancers?			Line 199. The authors need to define “pre-cancer” in materials.	1	2	medsci4030012_makarova	0
+7) Lines 238-242 are replicated in discussion lines 376-380.			Please give more details. Lines 214-216. It is not clear which statistical method is used for the significant results.	1	2	medsci4030012_makarova	0
+It is not clear which statistical method is used for the significant results in Table 2.			Lines 238-242 are replicated in discussion lines 376-380. The authors may need to delete the part in lines 238-242.	1	2	medsci4030012_makarova	0
+Therefore, if the authors can state that their sample is essentially Mexican that would be more informative and precise.			My main issue is with the way the results are presented. The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value. Even if they performed a test, important questions remain on the selection of the reference. The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry. The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA. My suggestion is to offer a better background for these results by trying to answer two main questions: 1)	1	2	medsci4030012_perova	0
+Tobari and colleagues studied the CNV burden in patients with cervical cancer from Mexican American ancestry.			is the deletion burden observed really exceptional in a population sample of similar ancestry? To answer this question, I would refer to the 1000 genomes data where there are Mexicans, Colombians, Puerto Ricans and Peruvians. There must be available CNV estimations for these ancestries. If you find good estimates then perhaps a statistical tests with those as your control group would be more believable.	1	2	medsci4030012_perova	0
+They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies. Please cite those.	1	2	medsci4030012_perova	0
+Note that there are differences in admixture in the latino population depending of their country of origin.			The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract. I suggest bringing this result up to the surface and making it more relevant even from the abstract. You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.	1	2	medsci4030012_perova	0
+Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			I missed a definition of the sample's ancestry. In the title it refers to Mexican American but in the rest of the manuscript it is treated as a latino population. Note that there are differences in admixture in the latino population depending of their country of origin. Studies on the 1000 genomes show differences between Puerto Ricans, Colombians, Mexicans and Peruvians. Therefore, if the authors can state that their sample is essentially Mexican that would be more informative and precise. I would then limit the use of latino. However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.	1	2	medsci4030012_perova	0
+The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry.			Abstract Line 23: should read Latino Populations	1	2	medsci4030012_perova	0
+2- The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract.			Figure 1: I cannot see the supposed deletions marked in green	1	2	medsci4030012_perova	0
+Then again there must be estimates of deletion burden for different types of cancers from TCGA studies.			Table 2b: Please remove the % sign in some of the table cells, for instance in Reference (Deletion, 10-100kb).	1	2	medsci4030012_perova	0
+My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			1) Line 34. The authors need to make clear about “pre-, cancer (91.3%) groups”. It is not clear if there is one or two groups but just one percentage.	1	2	medsci4030012_perova	0
+The results are novel and provide insight into how multiple CNAs might contribute to cervical cancer development.			2) Line 40. The authors may add up to 6 key words.	1	2	medsci4030012_perova	0
+My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			3) Lines 196-197. It is not clear which statistical method is used for the significant results in Table 2.Please give more details.	1	2	medsci4030012_perova	0
+Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			4) Line 199. The authors need to define “pre-cancer” in materials.	1	2	medsci4030012_perova	0
+The authors need to make clear about “pre-, cancer (91.3%) groups”.			Please give more details. 6) Lines 214-216. It is not clear which statistical method is used for the significant results.	1	2	medsci4030012_perova	0
+Recurrent aberrations of 98 CNA regions were also identified in cases only.			7) Lines 238-242 are replicated in discussion lines 376-380. The authors may need to delete the part in lines 238-242.	1	2	medsci4030012_perova	0
+The rationale for undertaking the research is a bit puzzling.			There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.	1	2	molecules26247651_makarova	0
+Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.			2. Full characterization of all synthesized compounds is required in the experimental section.	1	2	molecules26247651_makarova	0
+Check the font of the reference numbers in the text and correct			3. All the 1HNMR and 13CNMR must be attached as supplementary information.	1	2	molecules26247651_makarova	0
+All the 1HNMR and 13CNMR must be attached as supplementary information.			4. Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.	1	2	molecules26247651_makarova	0
+In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			5. The cell values do not align please correct them.	1	2	molecules26247651_makarova	0
+It is a continuation of similar OH...O binding studies performed 10 years ago.			6. Use 4 significant figures for the RNH values in table 2.	1	2	molecules26247651_makarova	0
+There are some questions about the manuscript as follows: 1.			7. It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.	1	2	molecules26247651_makarova	0
+The rationale for undertaking the research is a bit puzzling.			8. Check the font of the reference numbers in the text and correct RESPONSE:	1	2	molecules26247651_makarova	0
+Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			Correct subscripts and superscripts in the experimental also check the spacings to be consistent.	3	2	molecules26247651_makarova	0
+I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			In this article, the authors describe the spectral characteristics of NH in the hydrogen bridge NH...O. It is a continuation of similar OH...O binding studies performed 10 years ago. The rationale for undertaking the research is a bit puzzling. I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O. The explanation should be cleared.	1	2	molecules26247651_makarova	0
+The explanation should be cleared.Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major: 1) The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.			The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.	1	2	molecules26247651_makarova	0
+I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			Performing calculations with the Grimme dispersion correction did not make sense here, as D3 is important for intermolecular interactions, especially stacking. Here, calculations are performed for isolated molecules. Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.	1	2	molecules26247651_makarova	0
+Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major: 1)			183: should be SD = 34.3 cm-1	1	2	molecules26247651_makarova	0
+Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major: 1)			"186: ""are reasonable"": Taking into account that (i) this fit is rather unphysical (ii) moving the points by say 50 cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."	1	2	molecules26247651_makarova	0
+The cell values do not align please correct them.			Figure 5: Why is the y-axis reversed?	1	2	molecules26247651_makarova	0
+It is a continuation of similar OH...O binding studies performed 10 years ago.			1. There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.	1	2	molecules26247651_perova	0
+There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.2.			2. Full characterization of all synthesized compounds is required in the experimental section.	1	2	molecules26247651_perova	0
+The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			3. All the 1HNMR and 13CNMR must be attached as supplementary information.	1	2	molecules26247651_perova	0
+There are some questions about the manuscript as follows:1.			4. Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.	1	2	molecules26247651_perova	0
+- The ‘delay’ of 10 years is due to technical and personal factors of no relevance to the reported results.			5. The cell values do not align please correct them.	1	2	molecules26247651_perova	0
+The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			6. Use 4 significant figures for the RNH values in table 2.	1	2	molecules26247651_perova	0
+Full characterization of all synthesized compounds is required in the experimental section.			7. It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.	1	2	molecules26247651_perova	0
+8. Check the font of the reference numbers in the text and correct			8. Check the font of the reference numbers in the text and correct RESPONSE:	1	2	molecules26247651_perova	0
+There are some questions about the manuscript as follows:			Correct subscripts and superscripts in the experimental also check the spacings to be consistent.	3	2	molecules26247651_perova	0
+I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			1) The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.	1	2	molecules26247651_perova	0
+Full characterization of all synthesized compounds is required in the experimental section.			2) Performing calculations with the Grimme dispersion correction did not make sense here, as D3 is important for intermolecular interactions, especially stacking. Here, calculations are performed for isolated molecules. Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.	1	2	molecules26247651_perova	0
+2) Performing calculations with the Grimme dispersion correction did not make sense here, as D3 is important for intermolecular interactions, especially stacking.			183: should be SD = 34.3 cm-1	1	2	molecules26247651_perova	0
+We have for some time wished to investigate the corresponding NH···O systems.			"186: ""are reasonable"": Taking into account that (i) this fit is rather unphysical (ii) moving the points by say 50 cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."	1	2	molecules26247651_perova	0
+- The ‘delay’ of 10 years is due to technical and personal factors of no relevance to the reported results.			Figure 5: Why is the y-axis reversed?	1	2	molecules26247651_perova	0
+Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method.			The manuscript by Sharmin Akther Rupa et al, the manuscript has many serious concerns on the basis that it cannot be accepted in this journal. for example, 13C NMR must be 100 MHz not 400 MHz, value must be either ascending order or descending order (13C NMR (400 MHz, DMSO-d6, ppm): 1 above example showed the manuscript not written carefully.	1	2	molecules27051656_makarova	0
+Acta, 1995, 240, 93-98. https://doi.org/10.1016/0020-1693(95)04648-8 [85] Pearson, R. G. Absolute electronegativity and hardness correlated with molecular orbital theory.			Second one that is very important for this paper: The activity of L2 molecule against Bacillus Megaterium (PDB 4j6u) was significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which reveals the antimicrobial activity of the molecule. Both compounds are biologically active, but their activity was moderate which did not support their efforts.	1	2	molecules27051656_makarova	0
+A. K.; Heath, S. L. Self-assembled polynuclear clusters derived from some flexible polydentate dihydrazide ligands.			4. The authors did not report any Tyrosinase enzyme inhibition assay, they are requested to justify Tyrosinase inhibition as the probable mechanism of action for these sets of ligands by providing suitable literature references.	1	2	molecules27051656_makarova	0
+[97] Kang, S. M.; Heo, S. J.; Kim, K. N.; Lee, S. H.; Yang, H. M.; Kim, A. D.; Jeon, Y. J.			In this present study wehave synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and massspectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also doneThis manuscript is suitable publication in Molecules.	3	2	molecules27051656_makarova	0
+The molecular study needs a control molecule to compare the results of L1 and L2.			The manuscript is well-written and must be accepted in its present form for publication in Molecules.	1	2	molecules27051656_makarova	0
+The revised parts in the manuscript are indicated by red characters.			The conclusion is missing.	1	2	molecules27051656_makarova	0
+M.; Riou, A.; Allain, M.; Khan, M. A.; Boue, G. M. Synthesis, structural and spectral studies of 5-methyl 2-furaldehyde thiosemicarbazone and its Co, Ni, Cu and Cd complexes.			Font of the reference is different from the text	1	2	molecules27051656_makarova	0
+We anticipate continued research regarding these classes of exciting organic ligands.			This sentence “‪Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development” should be better connected to the next one to explain why they are important for new drug development	1	2	molecules27051656_makarova	0
+[56] Mahmoudi, F.; Farhadi, S.; Dusek, M.; Poupon, M. Synthesis, Spectroscopy and X-ray Crystallography Structure of Pyridine 4-Carbaldehyde Semicarbazone Schiff Base Ligand.			Replace “‪In addition to that, the compounds were tested in vitro bioassays against some Gram-negative and Gram-positive ‪bacteria and the fungus strain showing specially promising results for L2. Molecular docking methodology was used to study molecular behavior of L1 and L2 with Bacillus Megaterium to identify their binding interactions” with “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.‪ Response:	1	2	molecules27051656_makarova	0
+The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis.			Section 2.3.1. Reference for the used strains is required. Why are those specific strains used?	1	2	molecules27051656_makarova	0
+That’s why, to investigate and compare the antimicrobial activity of the synthesized compounds with in vitro data, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed.			Please change the info accordingly. ODB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium cristal structure.	1	2	molecules27051656_makarova	0
+Author Response Manuscript ID #: Molecules 1583723 Journal: Molecules A detailed point-by-point response to the reviewers' comments We would like to thank the reviewers for the comments and endorsements of our work.			Section 3.2. and 4.9. The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis. Divide the data into those 2 subheads and add more about the software used to perform the docking and the parameters used (Grid box, extensiveness …) Response:	1	2	molecules27051656_makarova	0
+[98] Nokinsee, D.; Shank, L.; Lee, V. S.; Nimmanpipug, P. Estimation of Inhibitory Effect against Tyrosinase Activity through Homology Modeling and Molecular Docking.			Section 4.8. There is no proper description of the results and no discussion in this part. Please revise - This sentence should be moved to the materials and methods or deleted from the section “‪Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (JCM-1652) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) ‪were used in this study.”	1	2	molecules27051656_makarova	0
+[97] Kang, S. M.; Heo, S. J.; Kim, K. N.; Lee, S. H.; Yang, H. M.; Kim, A. D.; Jeon, Y. J.			The molecular docking study is missing a control to properly analyse the results.	1	2	molecules27051656_makarova	0
+The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay.			Conclusion: Repeated abstract. The conclusion must remind the reader why the article was written in the first place and why it is important in the field. The conclusion should briefly give an insight into the obtained results and also the limitations.	1	2	molecules27051656_makarova	0
+Nonpolar hydrogen atoms, Gasteiger partial charges, rotatable bonds, and grid box with dimensions 66.57 × 58.25 × 84.98 Å3 created on the tyrosinase with the aid of Auto Dock Tools 1.1.2 and spacing of 0.3750 Å.			"The molecular study needs a control molecule to compare the results of L1 and L2. how would we know that −7.7 and −8.8 are good scores or not. Either perform a quick analysis for a molecule known for its tyrosinase inhibition or compare with other studies control that used the same methods in your paper.For exemple in this study ""https://doi.org/10.1016/j.bmc.2011.10.078"" they used Arbutin to compare the results of their studied molecule."	3	2	molecules27051656_makarova	0
+Dr. Md Abdul Majed Patwary Comilla University Author Response File: Author Response.pdf			The references are in a weird format containing only the author's name, journal name and date. article title and other data are missing. Reference style followed by the journalAuthor 1, A.B. ; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.	3	2	molecules27051656_makarova	0
+synthesis, spectral studies and in vitro anti-amoebic activity.			The manuscript by Sharmin Akther Rupa et al, the manuscript has many serious concerns on the basis that it cannot be accepted in this journal. for example, 13C NMR must be 100 MHz not 400 MHz, value must be either ascending order or descending order (13C NMR (400 MHz, DMSO-d6, ppm): 1 above example showed the manuscript not written carefully.	1	2	molecules27051656_perova	0
+The molecular study needs a control molecule to compare the results of L1 and L2.			Second one that is very important for this paper: The activity of L2 molecule against Bacillus Megaterium (PDB 4j6u) was significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which reveals the antimicrobial activity of the molecule. Both compounds are biologically active, but their activity was moderate which did not support their efforts.	1	2	molecules27051656_perova	0
+The computed FT-IR analysis as well as the 1H and 13C NMR using B3LYP/CC-PVDZ/6-311+G(d, p) method agreed satisfactorily with the experimental results.			4. 3. 2. Third one their binding study: 1. Why did the authors consider Tyrosinase from Bacillus megaterium N205A mutant pdb id 4j6u? As they did not report any enzyme specific inhibition related experiments. Docking is a preliminary experiment. How did the author validate the docking protocol? The authors are suggested using some decoy ligands and calculating the enrichment value in order to justify the docking protocol followed by ALA scan (Alanine scanning) based MD experiments. In addition, authors are suggested to report at list 100ns Molecular dynamics (MD) simulation guided protein-ligand stability report followed by either MM-GB or PB-SA based thermo data analysis. The pdb id 4j6u does not contain any bound ligand. How did the author select the binding site? If a few sets of amino acids were considered to define the binding site, then on what basis they select them? The authors did not report any Tyrosinase enzyme inhibition assay, they are requested to justify Tyrosinase inhibition as the probable mechanism of action for these sets of ligands by providing suitable literature references.	1	2	molecules27051656_perova	0
+Please be brief; most of the text is not important for a reader who only wants to get an idea about your paper before reading it.			"Abedin et al. explored ""Synthesis of Novel Tritopic Hydrazone Ligands: Spectroscopy, Biological activity, DFT, and Molecular docking Studies"". The manuscript is well-written and must be accepted in its present form for publication in Molecules."	1	2	molecules27051656_perova	0
+The binding site residues predicted by CastP for tyrosinase were used for grid generation.			The conclusion is missing.	1	2	molecules27051656_perova	0
+- This sentence “ ) — NHN=CH — CO — hydrazone moiety ( - containing hydrazide Polytopic ligands are important for new drug development” should be better connected to the next one to explain why they are important for new drug development?			Font of the reference is different from the text	1	2	molecules27051656_perova	0
+Design, synthesis, molecular docking and biological evaluation of imides, pyridazines, imidazoles derived from itaconic anhydride for potential antioxidant and antimicrobial activities.			This sentence “‪Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development” should be better connected to the next one to explain why they are important for new drug development	1	2	molecules27051656_perova	0
+Synthesis, structural, DFT studies, docking and antibacterial activity of a xanthene-based hydrazone ligand.			Replace “‪In addition to that, the compounds were tested in vitro bioassays against some Gram-negative and Gram-positive ‪bacteria and the fungus strain showing specially promising results for L2. Molecular docking methodology was used to study molecular behavior of L1 and L2 with Bacillus Megaterium to identify their binding interactions” with “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.‪ Response:	1	2	molecules27051656_perova	0
+An octanuclear [Co(II)2–Co(III)2]2 interlocked grid example of an inorganic [2]catenane†.			Reference for the used strains is required. Why are those specific strains used?	1	2	molecules27051656_perova	0
+It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].			Please change the info accordingly. ODB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium cristal structure.	1	2	molecules27051656_perova	0
+We also revise the whole manuscript to make the language and grammar better as suggested by the reviewer.			The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis. Divide the data into those 2 subheads and add more about the software used to perform the docking and the parameters used (Grid box, extensiveness …) Response:	1	2	molecules27051656_perova	0
+We also revise the whole manuscript to make the language and grammar better as suggested by the reviewer.			There is no proper description of the results and no discussion in this part.	1	2	molecules27051656_perova	0
+Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized topresent a wide range of biological activities in recent years.			This sentence should be moved to the materials and methods or deleted from the section “‪Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), ‪two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (JCM-1652) bacteria, ‪and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) ‪were used in this study.”	1	2	molecules27051656_perova	0
+Enzyme Research, 2015, 2015, 1-12. http://dx.doi.org/10.1155/2015/262364 [99] Cardoso, R.; Valente, R.; Souza da Costa, C.H.			The molecular docking study is missing a control to properly analyse the results.	1	2	molecules27051656_perova	0
+A.; Abu El‐Reash, G. M.; Bedier, R. A. Synthesis, spectroscopic, DFT, biological studies and molecular docking of oxovanadium (IV), copper (II) and iron (III) complexes of a new hydrazone derived from heterocyclic hydrazide.			"The molecular study needs a control molecule to compare the results of L1 and L2. how would we know that −7.7 and −8.8 are good scores or not. Either perform a quick analysis for a molecule known for its tyrosinase inhibition or compare with other studies control that used the same methods in your paper.For exemple in this study ""https://doi.org/10.1016/j.bmc.2011.10.078"" they used Arbutin to compare the results of their studied molecule."	3	2	molecules27051656_perova	0
+[42] Becke, A. D. Density‐functional thermochemistry.			The references are in a weird format containing only the author's name, journal name and date. article title and other data are missing. Reference style followed by the journalAuthor 1, A.B. ; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.	3	2	molecules27051656_perova	0
+This work presented the method for multiple metal ions detection using a carbon quantum dots (CQDs) based chemosensor array by functionalization with different amino acids.			line. 57-58. It seems that Figure 1 should be placed in other section, namely Section 3.1. , where it is mentioned for the first time (line 176).	1	2	molecules27123843_makarova	0
+This work presented the method for multiple metal ions detection using a carbon quantum dots (CQDs) based chemosensor array by functionalization with different amino acids.			"Line 59. ""Remove """"2."" ""Results"""""""	1	2	molecules27123843_makarova	0
+It is also impressive that all 11 metal ions were successfully identified by LDA with 100% classification rate.			line 148. What does NAA mean?	1	2	molecules27123843_makarova	0
+The CQDs with and without metal ions were systematically characterized (TEM, IR, XRD, XPS, etc).			"line 251. ""Write """"On TEM images """" instead of """"Transmission electrone microscopy can be observed..."""""""	1	2	molecules27123843_makarova	0
+Responses to Reviwers’ comments and the description of revisions in the revised manuscript We would sincerely thank the Review for the time and effort in carefully reading the manuscript and in preparing the review reports.			"line 284. ""Remove the word """"that"""""""	1	2	molecules27123843_makarova	0
+We truly appreciate your positive comments on our work, as well as for raising interesting points, which lead to the improvement of the manuscript.			line. 57-58. It seems that Figure 1 should be placed in other section, namely Section 3.1. , where it is mentioned for the first time (line 176).	1	2	molecules27123843_perova	0
+A detailed supplementary material is very helpful and useful for readers.			"Line 59. ""Remove """"2."" ""Results"""""""	1	2	molecules27123843_perova	0
+The CQDs with and without metal ions were systematically characterized (TEM, IR, XRD, XPS, etc).			line 148. What does NAA mean?	1	2	molecules27123843_perova	0
+Although the method of preparation of these sensors is not new, the implementation to chemosensor array is well presented.			"line 251. ""Write """"On TEM images """" instead of """"Transmission electrone microscopy can be observed..."""""""	1	2	molecules27123843_perova	0
+It seems that Figure 1 should be placed in other section, namely Section 3.1. , where it is mentioned for the first time (line 176).			"line 284. ""Remove the word """"that"""""""	1	2	molecules27123843_perova	0
+From authors: Victor F. Tarasenko and Dmitry A. Sorokin Institute of High Current Electronics E-mail: VFT@loi.hcei.tsc.ru			Of course, metal vapors, nano- and micro-particles can affect the characteristics of the discharge. This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc. Similar studies and comparisons with known experimental data could be carried out on these objects. In this connection, the use of high-voltage pulse discharges is not justified. The authors draw an analogy with the formation of sprites, then, in my opinion, the study would be more suitable for another journal related, for example, atmospheric research.	1	2	nano12040652_makarova	0
+From authors: Victor F. Tarasenko and Dmitry A. Sorokin Institute of High Current Electronics E-mail: VFT@loi.hcei.tsc.ru			Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow.	1	2	nano12040652_makarova	0
+Reply: This judgment was removed from the text of the manuscript.			In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.	1	2	nano12040652_makarova	0
+"For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."""			The introduction section could be improved. Some sentences are not well written or convey obvious ideas. Moreover, the aim of this work is to present an overview about the discharge behavior. The introduction would benefit from adding more content and detailed review overview to this research, such as particle lifting in electrostatic discharge , Turbulence effect, Mist-containing environment, Electrode materials.	1	2	nano12040652_makarova	0
+Reply: This judgment was removed from the text of the manuscript.			Please describe better your experimental system, such as ignition energy.	1	2	nano12040652_makarova	0
+In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material.			The choice of the electrode material should be clearly explained in the present paper.	1	2	nano12040652_makarova	0
+In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.			Please explain the mechanism of different color arcs produced by electrodes of different materials.	1	2	nano12040652_makarova	0
+We hope for a positive decision regarding the publication of this article in this journal.			The topic is timely and the results provided are interesting. However, most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only. A spectral analysis is given for one discharge only (discharge in air with a copper electrode): more quantitative results, such as a spectral analysis of the other discharges also, would be helpful for the comparison between the discharges produced in the lab and the ones observed in the atmosphere.	1	2	nano12040652_makarova	0
+On the other hand, we should note that with a voltage pulse duration from ones to tens of nanoseconds, the experimental conditions differ significantly from the conditions of an electrostatic discharge and the creation of a foggy environment.			A point that should be clarified is the choice of the gases used for the discharges. One would expect a comparison between discharges that happen in gas mixtures of similar composition. Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?	1	2	nano12040652_makarova	0
+The brightness of the micrometeorite (the particle) glow increases towards the track’s end.			Page 5, line 168 The authors state: “An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface.” The sense of this sentence is not completely clear to me. Where does the evaporation of the metal take place? It it took place on the particle surface, it should produce a reduction of its size, rather than an increase.	1	2	nano12040652_makarova	0
+Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?			Page 6, line 218 The authors state: “Particles of metal and its compounds with oxygen and nitrogen with a size of 500 nm and less are nonuniformly distributed on the surface of the slide”. Some data should be provided to show the elemental composition of the particles.	1	2	nano12040652_makarova	0
+Here are some of them: 1) Of course, metal vapors, nano- and micro-particles can affect the characteristics of the discharge.			Of course, metal vapors, nano- and micro-particles can affect the characteristics of the discharge. This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc. Similar studies and comparisons with known experimental data could be carried out on these objects. In this connection, the use of high-voltage pulse discharges is not justified. The authors draw an analogy with the formation of sprites, then, in my opinion, the study would be more suitable for another journal related, for example, atmospheric research.	1	2	nano12040652_perova	0
+Some sentences are not well written or convey obvious ideas.			"The article is more descriptive in nature, there is no explanation of physical laws, but which are misleading readers. For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."" One can agree with the influence of the particle size on the track size, but how can an increase in the particle charge affect the track glow? Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow. Such inaccuracies are found throughout the text."	1	2	nano12040652_perova	0
+Similar studies and comparisons with known experimental data could be carried out on these objects.			In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.	1	2	nano12040652_perova	0
+We hope for a positive decision regarding the publication of this article in this journal.			The introduction section could be improved. Some sentences are not well written or convey obvious ideas. Moreover, the aim of this work is to present an overview about the discharge behavior. The introduction would benefit from adding more content and detailed review overview to this research, such as particle lifting in electrostatic discharge , Turbulence effect, Mist-containing environment, Electrode materials.	1	2	nano12040652_perova	0
+In addition, there are comments on the research methodology and interpretation of research on the article itself.			Please describe better your experimental system, such as ignition energy.	1	2	nano12040652_perova	0
+One can agree with the influence of the particle size on the track size, but how can an increase in the particle charge affect the track glow?			The choice of the electrode material should be clearly explained in the present paper.	1	2	nano12040652_perova	0
+High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap.			Please explain the mechanism of different color arcs produced by electrodes of different materials.	1	2	nano12040652_perova	0
+Such inaccuracies are found throughout the text.			most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only. A spectral analysis is given for one discharge only (discharge in air with a copper electrode): more quantitative results, such as a spectral analysis of the other discharges also, would be helpful for the comparison between the discharges produced in the lab and the ones observed in the atmosphere.	1	2	nano12040652_perova	0
+Reply to Reviewer 2 The article: «Influence of nanoparticles and metal vapors on the color of laboratory and atmospheric discharges» Authors: Victor Tarasenko, Nikita Vinogradov, Dmitry Beloplotov, Alexander Burachenko, Mikhail Lomaev, and Dmitry Sorokin Submission Date: 14 December 2021 Date of this review: 19 Jan 2022 07:18:39 Comments and Suggestions for Authors 1.			A point that should be clarified is the choice of the gases used for the discharges. One would expect a comparison between discharges that happen in gas mixtures of similar composition. Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?	1	2	nano12040652_perova	0
+We note once again that in this work, to obtain metal vapors, as well as metal nano- and microparticles, a pulsed nanosecond discharge in a non-uniform electric field was used.			A couple of requests of clarification about specific parts of the text are also provided in the following. Page 5, line 168 The authors state: “An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface.” The sense of this sentence is not completely clear to me. Where does the evaporation of the metal take place? It it took place on the particle surface, it should produce a reduction of its size, rather than an increase.	1	2	nano12040652_perova	0
+However, most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only.			Page 6, line 218 The authors state: “Particles of metal and its compounds with oxygen and nitrogen with a size of 500 nm and less are nonuniformly distributed on the surface of the slide”. Some data should be provided to show the elemental composition of the particles.	1	2	nano12040652_perova	0
+As it was, I couldn't understand the description provided, and it didn't seem to match the figures, which were simply labeled as FM, so seemed to be a simple plots of FM using 2 methods on first read.			TITLE The “Part 1” in the title, while perhaps intriguing, calls for some explanation of future directions. This does not seem to be addressed in the article, thus there seems to be no reason to include this phrase in the title.	1	2	nu14051073_makarova	0
+Response to reviewers' comments: Reviewer one: TITLE The “Part 1” in the title, while perhaps intriguing, calls for some explanation of future directions.			Also, the difference(s) between FFM and LM definitions are nuanced and complex and should either be defined, or alternate text should be used here. It seems that the authors are simply trying to point out that use of the 4C model is not common and that other methods are normally used. Best to clarify this statement. INTRODUCTION P2 L61: FFM is obtained via the 4C model – it is not a surrogate.	1	2	nu14051073_makarova	0
+of the paper is all about comparing techniques against the 4C model.			P2 L69: The aim stated here does not follow the logic of the preceding statements in the paragraph, i.e., what does comparing BC methods have to do with lack of healthcare or technology? Is the focus then on finding low cost, easy to apply methods? The aim (and intent) of the study seems too simplistic as stated here – merely comparing methods. How would the results be applied to the healthcare situation in Mexico? These topics should be tied together or an alternative motivation needs to be presented.	1	2	nu14051073_makarova	0
+Reviewer two comments: I am not familiar with the Bland-Altman method, so was unclear when I read the methods section whether the text in lines 204-207 was explaining the Bland-Altman method or describing a separate procedure.			Who was asked to assent? MATERIALS AND METHODS P3 L94: This is awkwardly worded.	1	2	nu14051073_makarova	0
+I hope that you find this work suitable for your Journal and I look forward to hearing from you.			P3 L125: DXA: Was the head ROI excluded from the analyses (as per recommendation by the ISCD)? If so, it should be stated. If not, DXA analysis should be redone and all relationships recalculated.	1	2	nu14051073_makarova	0
+I didn't understand why the data differed between Table 1 and Supplementary Table 1.			P5 L176: “(n=52?? )” Is there uncertainty of the number of participants or is this merely a typo?	1	2	nu14051073_makarova	0
+P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).			P5 L207: The means of FM would be compared using t-test, not calculated.	1	2	nu14051073_makarova	0
+Table 1: The n values in the column headers add up to 293 rather than 288.			Are the n values incorrect or is the entire cohort presented here? RESULTS Table 1: The n values in the column headers add up to 293 rather than 288.	1	2	nu14051073_makarova	0
+Please find our answers in the following lines.			Table 1; Body Composition Variables: The mean values do not all agree with those in supplementary Table 1.	1	2	nu14051073_makarova	0
+MATERIALS AND METHODS P3 L94: This is awkwardly worded.			Table 1; MRI subsample: There is no indication of age, BMI, etc. in this group. That information may be helpful, perhaps also as supplementary data.	1	2	nu14051073_makarova	0
+over or under estimating FM) may affect health assessments.			P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).	1	2	nu14051073_makarova	0
+If the focus (see the title) of the article is comparisons with the 4C model, then supplementary Table 1 seems to merit more direct attention.			P12 L283: Is Table 4 necessary? The title (and aim?) of the paper is all about comparing techniques against the 4C model. One also begins to wonder about affecting type 1 errors due to multiplicity of comparisons (debatable but worth considering). It may be worth considering moving Table 4 to the supplementary file and bringing supplementary Table 1 into the main document. If the focus (see the title) of the article is comparisons with the 4C model, then supplementary Table 1 seems to merit more direct attention.	1	2	nu14051073_makarova	0
+MATERIALS AND METHODS P3 L94: This is awkwardly worded.			As stated above, it may be worth considering moving these Bland-Altman figures to the supplementary file.	1	2	nu14051073_makarova	0
+Concluding that methods differ is not surprising.			It should perhaps be the last paragraph in the Discussion. DISCUSSION P15 L393: This paragraph reads more like a conclusion and seem out of place.	1	2	nu14051073_makarova	0
+"(E.g., ""A positive trend indicates increasing underestimation of FM at high FM levels; a negative trend indicates increasing overestimation of FM at high FM levels."			Much of the Discussion addresses strengths and weaknesses of the different BC methods, as if to provide guidance for clinicians in selecting the best techniques according to their needs and capabilities. None of the methods are ideal – all are flawed (in comparison with a gold standard). Yet a firm conclusion seems to be lacking. Concluding that methods differ is not surprising. One may be better served by discussing how the differences in methods (i.e. over or under estimating FM) may affect health assessments. That is, what impact may underestimating FM, by relying on one particular method, have on child care? How critical is the method selection? Somehow this needs to be tied more firmly to the population and environment being studied.	1	2	nu14051073_makarova	0
+P3 L125: DXA: Was the head ROI excluded from the analyses (as per recommendation by the ISCD)?			"I am not familiar with the Bland-Altman method, so was unclear when I read the methods section whether the text in lines 204-207 was explaining the Bland-Altman method or describing a separate procedure. I take from the results that those sentences were describing the Bland-Altman procedure, and if so, adding some text along the lines of ""In this procedure..."" would be helpful. As it was, I couldn't understand the description provided, and it didn't seem to match the figures, which were simply labeled as FM, so seemed to be a simple plots of FM using 2 methods on first read."	1	2	nu14051073_makarova	0
+Are the n values incorrect or is the entire cohort presented here?			"Relatedly, it would have been helpful to have more explanatory titles for the plots--eg ""Differential correlation between methods across levels of FM"". And, the meaning of the trend line was very counter-intuitive, so an interpretive note under the plots would have been helpful. (E.g., ""A positive trend indicates increasing underestimation of FM at high FM levels; a negative trend indicates increasing overestimation of FM at high FM levels."	1	2	nu14051073_makarova	0
+It seems that the authors are simply trying to point out that use of the 4C model is not common and that other methods are normally used.			There is no description of the race and ethnic composition of the sample. I am not well-versed in the ethnic composition of Mexico, however, would it be helpful to know the degree of representation of, for example, youth who are Black, of Indigenous ancestry, and of European ancestry? Or are there other ethnic or cultural groups that should be represented?	1	2	nu14051073_makarova	0
+Is the focus then on finding low cost, easy to apply methods?			I didn't understand why the data differed between Table 1 and Supplementary Table 1. Why did Ns (and means/SDs) differ?	1	2	nu14051073_makarova	0
+OVERALL Much of the Discussion addresses strengths and weaknesses of the different BC methods, as if to provide guidance for clinicians in selecting the best techniques according to their needs and capabilities.			Not all abbreviations are listed for the table footnotes (FFM, D2O, DXA, LM, BV, ADP)	1	2	nu14051073_makarova	0
+The following paper did a better job of reporting it Moraeus L, Lindroos AK, Warensjö Lemming E, Mattisson I.			Abstract: it is not conventional. It is very “fragmented”. Try to elaborate it again.	1	2	nu14122489_perova	0
+Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet.			Theoretical framework: it is very updated. To be prudent, try to update some references if you find.	1	2	nu14122489_perova	0
+Given the findings the researchers could be arguing that a review may be needed with the Swedish school lunch program and increase the level of fruit and vegetables in the students' diet, particularly for students in lower SES locations.			Methods. This paper is based on cross sectional method. Can you justify it in depth? This study includes a size of 1139 adolescents.	1	2	nu14122489_perova	0
+The first sentence assumes that poor diet is always associated with adolescents, this is incorrect, Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing			Results. In Results, the separations of text and Figures and Tables is strange (when a paper does it, the trend is to include it at the end of all the paper). I recommend you to alternate them (text, Figures and Tables in Results). Nevertheless, results are well elaborated.	1	2	nu14122489_perova	0
+The lack of recommendations associated with the Swedish free lunch program to assist the diets of students in low SES families to have more fruit and vegetables			Conclusion and discussion: They are well elaborated.	1	2	nu14122489_perova	0
+A public education program could also be encouraged about health eating.			The level of reporting of the core assessment instruments and their items, dimensions and subdomains needs to be enhanced.	1	2	nu14122489_perova	0
+It needs minor revision in places but the results need more attention.			There is a need to revising the reporting of the results and information in the tables along with the statistical methodology used with interval data and group comparison research.	1	2	nu14122489_perova	0
+It needs minor revision in places but the results need more attention.			The focus on only the global mean score for the wellness KQ-10 measure rather than the 5 dimensions. Diet is only expected to influence 2 of these 5 dimensions.	1	2	nu14122489_perova	0
+There is a need to revising the reporting of the results and information in the tables along with the statistical methodology used with interval data and group comparison research.			The lack of recognition that income and cost of living rather than education of parents is also a likely reason for the findings.	1	2	nu14122489_perova	0
+If income is not important than the paper should be the on home education level and home diet not home SES and home diet.			The lack of recommendations associated with the Swedish free lunch program to assist the diets of students in low SES families to have more fruit and vegetables	1	2	nu14122489_perova	0
+Effects of socioeconomic status on nutrition in Asia and future nutrition policy studies.			The first sentence assumes that poor diet is always associated with adolescents, this is incorrect Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing.	1	2	nu14122489_perova	0
+Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			Line 90 need to put in the full name Health Related Quality of Life (HRQoL). You have so many abbreviations confusing to follow in places. The authors may know what these are, but many reader will not. To assist the reader’s comprehension, explain the instruments more as you develop the paper.	1	2	nu14122489_perova	0
+Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			The important issue is in the method section the reader needs understand the dimensions and sub-dimension that make up within each survey. Unless these are included the reader can not fully understand the study.	1	2	nu14122489_perova	0
+Schools may be making the lunches to a budget rather than to a healthy diet criteria which is more expensive.			The term is usually SES social economic status so it should be SES- F if it is social economic status – factors. Given you are using education of parents as the status measure of SES the term status is important in this paper and should not be dropped.	1	2	nu14122489_perova	0
+-Conclusion and discussion: They are well elaborated.			It is more that education it is also income re line 106-107 “It is proposed that parents with more educational experience are more likely to make healthier food choices for their family” . Miss the point It also assumes that parents with more educational experience are more likely to have higher incomes and so are more likely to make and avoid healthier food choices for their family	1	2	nu14122489_perova	0
+You may have a more important study if you do that, with a different finding to what you have just using the composite total.			It also assumes that parents with more educational experience are more likely to have higher incomes and so are more likely to make and avoid healthier food choices for their family.	1	2	nu14122489_perova	0
+Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			If income is not important than the paper should be the on home education level and home diet not home SES and home diet.	1	2	nu14122489_perova	0
+It needs minor revision in places but the results need more attention.			See Murayama, N. (2015). Effects of socioeconomic status on nutrition in Asia and future nutrition policy studies. Journal of nutritional science and vitaminology, 61(Supplement), S66-S68. Darmon, N., & Drewnowski, A. (2008). Does social class predict diet quality?. The American journal of clinical nutrition, 87(5), 1107-1117. The issues raised in the well quoted Darmon and Drewnowski paper need to be considered more.	1	2	nu14122489_perova	0
+The American journal of clinical nutrition, 87(5), 1107-1117.			The Swedish Healthy Eating Index for Adolescents 2015 (SHEIA15) is not well explained in this paper. The following paper did a better job of reporting it Moraeus L, Lindroos AK, Warensjö Lemming E, Mattisson I. Diet diversity score and healthy eating index in relation to diet quality and socio-demographic factors: results from a cross-sectional national dietary survey of Swedish adolescents. Public Health Nutr. 2020 Jul;23(10):1754-1765. doi: See their Table 5.	1	2	nu14122489_perova	0
+The European Journal of Health Economics, 15(9), 967-977.			The KIDSCREEN-10 is not well described. The basic information is not there in the paper: “KIDSCREEN-10 (KS-10) is derived from the KIDSCREEN-27, and provides a single index of global QoL using ten items related to physical well-being, psychological well-being, autonomy and parent relation, social support and peers, and school environment” See how these researchers have described the KIDSCREEN-10 . Bouwmans, C., van der Kolk, A., Oppe, M., Schawo, S., Stolk, E., van Agthoven, M., ... & van Roijen, L. (2014). Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD. The European Journal of Health Economics, 15(9), 967-977.	1	2	nu14122489_perova	0
+Given the findings the researchers could be arguing that a review may be needed with the Swedish school lunch program and increase the level of fruit and vegetables in the students' diet, particularly for students in lower SES locations.			In terms of results the flowchart was interesting.	1	2	nu14122489_perova	0
+In table 5 only one p value is reported, but what it is measuring is unclear, as there are a number of interactions occurring.			I am a not European reviewer and so found the RADDS a rather restricted list without some meat.	1	2	nu14122489_perova	0
+You may have a more important study if you do that, with a different finding to what you have just using the composite total.			Why was the Pearson's chi-squared test used (table 1) for analysis of variance? For while gender and in this study education are categorical (group) the data being evaluated is continuous and interval data and so an ANOVA or MANOVA by group is the method of analysis of variance. (Tabachnick, B.G., Fidell, L. S., & Ullman, J.B. (2007). Using multivariate statistics (5th ed.). Pearson.)	1	2	nu14122489_perova	0
+If income is not important than the paper should be the on home education level and home diet not home SES and home diet.			In terms of gender and education: the mean, standard deviation, df and sig t or F test need to be reported in the tables.	1	2	nu14122489_perova	0
+Use all five of the KO-10 dimensions as your outcome measure not the Mean average global KQ-10 score.			Similarly, a correlation matrix is typically reported as it the foundation of regression analyses and so it needs to be reported to understanding the interaction between three main tests variables being investigated in this study.	1	2	nu14122489_perova	0
+Remember the KQ-10 is made up of five subdomains (psychological well-being, autonomy and parent relation, social support and peers, and school environment) The fact is diet is no expected to change 3 of these KQ-3 subdomains and only one or two KQ-10 dimensions.			The regression analyses “p” value is reported, but the beta values and significance must also be reported. The regressing table needs reworked, as it is the influence of diet the independent variable on wellbeing KO-10, the dependent variable. If the focus is on gender typically both a boy and a girl regression model is reported.	1	2	nu14122489_perova	0
+I am a not European reviewer and so found the RADDS a rather restricted list without some meat.			Table 5 is interesting but the setting out is poor and so the columns do not align with headings, particularly the wellness KQ -10 information. Again, an ANOVA “ t” value and df as well as the p value have to be reported. In table 5 only one p value is reported, but what it is measuring is unclear, as there are a number of interactions occurring. Should be reporting total, then girls, and then boys as there look to be interaction effects.	1	2	nu14122489_perova	0
+Should be reporting total, then girls, and then boys as there look to be interaction effects .			Because the KQ-10 is a composite tests there is the likelihood to be some interaction with the sub-domains. Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet. Diet is not expected to have any influence of parent relations, social relations or peers but your study may find an influence on psychological well-being and even school environment. This is the core of your study: does diet have an influence on psychological wellbeing? Remember the KQ-10 is made up of five subdomains (psychological well-being, autonomy and parent relation, social support and peers, and school environment) The fact is diet is no expected to change 3 of these KQ-3 subdomains and only one or two KQ-10 dimensions. Use all five of the KO-10 dimensions as your outcome measure not the Mean average global KQ-10 score. You may have a more important study if you do that, with a different finding to what you have just using the composite total.	1	2	nu14122489_perova	0
+Diet is not expected to have any influence of parent relations, social relations or peers but your study may find an influence on psychological well-being and even school environment.			The conclusion is sound given the findings but the lack of reference to income is an issue as educational status of parents is often a “de-facto” measure for income. It maybe, it is the cost of living associated with fresh fruit and vegetables that is the real issue. Given the findings the researchers could be arguing that a review may be needed with the Swedish school lunch program and increase the level of fruit and vegetables in the students' diet, particularly for students in lower SES locations. Schools may be making the lunches to a budget rather than to a healthy diet criteria which is more expensive. Different sub-populations even in the same school may need different mix of foods. A public education program could also be encouraged about health eating.	1	2	nu14122489_perova	0
+Author Response Reviewer 3: General comments: The manuscript reports a study about weight-biased language in the Australian media across the last decades.			The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach. This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI. This study deals with question that needs important discussion for obesity-related public health policy development.	1	2	obesities2010010_makarova	0
+I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance.			This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.	1	2	obesities2010010_makarova	0
+The topic is interesting, and the manuscript is clear.			I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance. Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).	1	2	obesities2010010_makarova	0
+I do not have particular concerns about the paper.			Legend for Figure 2 is missing.	1	2	obesities2010010_makarova	0
+The topic is interesting, and the manuscript is clear.			Please move the research questions from the methods to the introduction.	1	2	obesities2010010_makarova	0
+I do not have particular concerns about the paper.			Last paragraph of the methods: why did you report here your conclusions? Please move this part from this section.	1	2	obesities2010010_makarova	0
+The manuscript reports a study about weight-biased language in the Australian media across the last decades.			Please revised Figure 1 because it's very difficult to read.	1	2	obesities2010010_makarova	0
+Is there any possible limit with this approach in the selection of the papers?			I think this aspect is interesting and linked to your results, showing a connection with clinical data. A recent paper has pointed out that postbariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z).	1	2	obesities2010010_makarova	0
+The revised version of the manuscript is appropriate for publishing.			You report that an automatic approach was used to check all the papers. Is there any possible limit with this approach in the selection of the papers?	1	2	obesities2010010_makarova	0
+You report that an automatic approach was used to check all the papers.			The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach. This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI. This study deals with question that needs important discussion for obesity-related public health policy development.	1	2	obesities2010010_perova	0
+I think this aspect is interesting and linked to your results, showing a connection with clinical data.			This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.	1	2	obesities2010010_perova	0
+I think this aspect is interesting and linked to your results, showing a connection with clinical data.			I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance. Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).	1	2	obesities2010010_perova	0
+I do not have particular concerns about the paper.			Legend for Figure 2 is missing.	1	2	obesities2010010_perova	0
+Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).			Please move the research questions from the methods to the introduction.	1	2	obesities2010010_perova	0
+A recent paper has pointed out that postbariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z).			Last paragraph of the methods: why did you report here your conclusions? Please move this part from this section.	1	2	obesities2010010_perova	0
+I have some comments for the authors about the structure of the manuscript that should take into consideration.			Please revise Figure 1 because it's very difficult to read.	1	2	obesities2010010_perova	0
+The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			A recent paper has pointed out that post-bariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z). I think this aspect is interesting and linked to your results, showing a connection with clinical data.	1	2	obesities2010010_perova	0
+Author Response Reviewer 1: General comments:			You report that an automatic approach was used to check all the papers. Is there any possible limit with this approach in the selection of the papers?	1	2	obesities2010010_perova	0
+The authors should consider reshaping the tables, eliminating unnecessary columns (e.g.			The main issue with the ms is the quality of the figures: they are not clearly representing the data, arrows and text may help the reader, the controls are missing in most cases, and a general lack of precision is affecting them. Also, the legends are not clearly describing all the terms and should be revised. Legends should describe what is to be observed in the figure, possibly pointing to parts of special interest.	1	2	plants11050622_makarova	0
+L127: Please add the explanation of “boro season”.			Explaining briefly which and what are the indian sesasons during which experiments have been conducted would help clarity.	1	2	plants11050622_makarova	0
+The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			"The phenotypic data are important, but currently they are presented only as additional material. The authors should consider reshaping the tables, eliminating unnecessary columns (e.g. no. of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion. Also ""entry nos."" is probably not clear and ILs no should maybe considered instead."	1	2	plants11050622_makarova	0
+is probably not clear and ILs no should maybe considered instead.			The data on bgs are not sufficiently presented in the text nor discussed.	1	2	plants11050622_makarova	0
+It is also acknowledged that this paper is probably the first of many papers to emerge from the study.			"revise the use of ""-"" instead of ""( )"" when mentioning the numbers of the ILs, or any other system which would ensure a better homogeneity in comparison to the one present in the ms."	1	2	plants11050622_makarova	0
+The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			check lines 21, 37, 93, 153, 183-184, 312, 351-353, 360, 377, 466	1	2	plants11050622_makarova	0
+Dear Editor, Thank you for inviting me to review this manuscript.			stick to BB instead of BLB	1	2	plants11050622_makarova	0
+The phenotypic data are important, but currently they are presented only as additional material.			consider anticipating the explanation of the acronyms which are eventually presented only in the M&M, to facilitate the reading; consider this aspect also in the legends	1	2	plants11050622_makarova	0
+Author Response Point-by-point response to reviewer's comments			Lines 403 et seq. are redundant Response:	1	2	plants11050622_makarova	0
+Then the table can be more informative and ease to read.			lines 414 et seq. seem to be contradicting lines 351 et seq.	1	2	plants11050622_makarova	0
+L48-63: should add some citations on the first part of introduction.			the 85 ILs presented at 414 seem to be a bit out of the blue: a short intro to where they are coming from would help the reader.	1	2	plants11050622_makarova	0
+The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			Thank you for inviting me to review this manuscript. The paper itself is well written, although 1) somewhat results are partially descriptive and partially inferential. However, the authors have conducted a thorough literature review, undertaken a rigorous piece of data collection, and have generalized information accurately.	1	2	plants11050622_makarova	0
+L127: Please add the explanation of “boro season”.			With minor grammatical revisions, the manuscript can be accepted as is.	1	2	plants11050622_makarova	0
+With minor grammatical revisions, the manuscript can be accepted as is.			It is also acknowledged that this paper is probably the first of many papers to emerge from the study. As such, it is an overview paper that raises many questions. It would be interesting for the authors to provide more information about the research design and estimation methodologies, such as chi-square analysis of introgression lines, if possible for each crossed IL population. I only recommend some minor revisions before acceptance. Response: Authors once again thank the reviewer for correct assessment of the basic purpose of the manuscript. Yes, this is truly an overview paper presenting the interesting observations from our study. The introgression scheme and phenotyping of various biotic traits and drought phenotyping have been described in detail under ‘Materials and Methods’ section. The metric data on yield traits was statistically analyzed and results of ANOVA, heritability and critical differences have been presented in the manuscript. Authors agree that it would be more inferential with chi square values. However, the crossing scheme was viewed holistically and data on each cross was not maintained separately as the present study aimed at selecting introgression lines from multiples crosses with multiple stress resistance/tolerance by pooling several genes and QTLs into a common background. Despite maintaining large base populations, plants per se were selected based on marker positivity for inter-crossing and selfing and further stringent phenotypic selection for the targeted traits. Hence, chi-square which is perfectly apt for population derived from biparental crosses is not used in our study. This work is of outstanding quality, and I normally present more critical points in my reviews. However, this time it is just very beautiful work.	1	2	plants11050622_makarova	0
+For example on L203 “CD”, L211 “SES“, L228 “UBN“ and L332 “ICAR-IRRR”.			Line 262: “=261 %” , and Line 312 “linkes” a typo? Correct it Response:	1	2	plants11050622_makarova	0
+The phenotypic data are important, but currently they are presented only as additional material.			It was a pleasure to read this manuscript. I wish the author of the best.	1	2	plants11050622_makarova	0
+The work is not unprecedented in the concept, but provides interesting breeding material for specific indian farmers, which is an important achievement and is correctly taken up by authorities and international research centers.			This paper described the multi-parents introgression assisted by molecular markers. The content is informative, however, the tables should be reorganized and the statistical methods description have to be improved. Please see the suggestion and comments below.	1	2	plants11050622_makarova	0
+L562: the section of statistical analysis should add more details.			L48-63: should add some citations on the first part of introduction.	1	2	plants11050622_makarova	0
+the 85 ILs presented at 414 seem to be a bit out of the blue: a short intro to where they are coming from would help the reader.			L127: Please add the explanation of “boro season”.	1	2	plants11050622_makarova	0
+However, this time it is just very beautiful work.			The legend of supplementary table should be improved. Table and main text are independent, so the authors have to describe the table more carefully.	1	2	plants11050622_makarova	0
+Then the table can be more informative and ease to read.			L562: the section of statistical analysis should add more details. Also please indicate the R version. For example, how does H2 calculate?	1	2	plants11050622_makarova	0
+It is also acknowledged that this paper is probably the first of many papers to emerge from the study.			On Supplementary table S2, does significant mean p-value? Indicate DFF = days to fifty percent flowering. What is the “treatment”? What do the results on “check” mean? How did you analyze “control vs IL”?	1	2	plants11050622_makarova	0
+Correct it It was a pleasure to read this manuscript.			On Supplementary table S3, similar questions as S2, please also explain.	1	2	plants11050622_makarova	0
+is probably not clear and ILs no should maybe considered instead.			On Supplementary table S5, what is “C.D”? How did you calculate those values on those comparison types?	1	2	plants11050622_makarova	0
+Also, the legends are not clearly describing all the terms and should be revised.			Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups. Then the table can be more informative and ease to read.	1	2	plants11050622_makarova	0
+Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups.			Please add the full name of abbreviations. For example on L203 “CD”, L211 “SES“, L228 “UBN“ and L332 “ICAR-IRRR”.	1	2	plants11050622_makarova	0
+Author Response Point-by-point response to reviewer's comments			The last paragraph of results is about background selection. Suggest to add subtitle “2.2.8”.	1	2	plants11050622_makarova	0
+of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion.			Please add some gel pictures of the foreground selection markers you used in order to visualize the genotyping results and showed the polymorphism of these markers on gel.	1	2	plants11050622_makarova	0
+This paper described the multi-parents introgression assisted by molecular markers.			L296-L302: the marker descriptions on main text cannot match the supplementary table S8. Also, what are those 27 ILs on L298?	1	2	plants11050622_makarova	0
+How did you calculate those values on those comparison types?			L602: there is no appendix.	1	2	plants11050622_makarova	0
+Our responses to your comments are as follows.			In general, the manuscript refers often to PGRMC1 expression or downregulation. These are fuzzy terms for proteins. It is much preferable to refer to protein abundance, and changes in abundance. Such changes may occur at the translational or post-translational level. Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool. The authors are strongly advised to change from e.g. ‘expression’ to ‘abundance’ throughout.	1	2	reprodmed3020015_makarova	0
+Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented. Therefore, the reviewer assumes that experiments have only been performed once. The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.	1	2	reprodmed3020015_makarova	0
+SEM is a function of how many measurements were made, and does not reflect sample variability.			Line 24. AG-205. The authors must refer to the fact that AG-205 is not specific for PGRMC1, contrary to many citations in the literature (such as would be the case here, if uncorrected). Cite PMID: 34944026, PMID: 34680104, PMID: 32924377. Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.	1	2	reprodmed3020015_makarova	0
+Furthermore, the required experiment is technically not challenging.			It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).	1	2	reprodmed3020015_makarova	0
+Unfortunately, the authors have been unable to demonstrate the direct involvement of PGRMC1 downregulation in decidualization.			Line 50. “(10-12); however,” Two discrete sentences “(10-12).	1	2	reprodmed3020015_makarova	0
+The statistical significance of any result would be evaluated using multivariate analysis.			Lines 76-78. Statistical sample sizes were unfavourably small. The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).	1	2	reprodmed3020015_makarova	0
+PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491).			Lines 88-98. Cell culture. What characterization was performed to confirm the identity of cells as ESCs? In general, there is very little detail on these primary cultured lines. How long are they held in culture? Are they stored cryogenically? How are they re-seeded into culture?	1	2	reprodmed3020015_makarova	0
+Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented.			Lines 121-133. What percentage acrylamide were the SDS PAGE gels. For what length of time and at what voltage settings was electrophoretic transfer to PVDF? In what device?	1	2	reprodmed3020015_makarova	0
+1) I requested a major revision including the use of miRNA-resistant exogenous PGRMC1 expression (altered codon usage) to demonstrate that lack of PGRMC1 attenuation will prevent decidualization.			Line 126-7. Give the catalogue number of the secondary antibody.	1	2	reprodmed3020015_makarova	0
+The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			Line 131. On what device was ECL light detected? What were the device settings? How was pixel saturation dealt with?	1	2	reprodmed3020015_makarova	0
+Sabbir’s results (ref 31) imply that this also holds for PGRMC1.			Line 13 Which ImageJ plug-in was used for quantification?	1	2	reprodmed3020015_makarova	0
+PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491).			Lines 134-137. Result errors should not be expressed as SEM, but as standard deviation (s.d.). All results should show s.d. error bars.	1	2	reprodmed3020015_makarova	0
+In Fig.4 the authors argue that decidualization markers are induced by both db-cAMP and db-cAMP/P4, and that therefore P4 is not involved in the PGRMC1 response.			Line 150. The legend to fig. 1. inappropriately ends with “2.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.	1	2	reprodmed3020015_makarova	0
+Appropriate human ethics approvals were obtained.			Lines 153-156, Figure 2. The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient. This result must be demonstrated in a substantively convincing manner.	1	2	reprodmed3020015_makarova	0
+If further mechanism exploration is carried out, the research will be more complete and persuasive.			2. Line 158/Fig. The western blot (also Fig.3) result must show the entire molecular weight range (e.g. in supplemental data), including MW markers. PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491). The latter could well contribute to alterations in PGRMC1 abundance cited by this study. It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.	1	2	reprodmed3020015_makarova	0
+Appropriate human ethics approvals were obtained.			Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true). Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels. In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization. It is important to determine whether these mechanisms are both operating. At the very least this mechanism merits detailed discussion in that section. PGRMC1 protein instability could be the dominant effect involved, with miRNA transcript regulation playing only a minor part.	1	2	reprodmed3020015_makarova	0
+Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) Lines 271-76.			Line 163. Change 3.2. to 3.3.	1	2	reprodmed3020015_makarova	0
+It is important to determine whether these mechanisms are both operating.			Lines 164-172. This cell culture experiment also seems to be based upon just one primary cell culture, split into replicates. I am uncertain because the methods do not explain how the experiment was performed. Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.	1	2	reprodmed3020015_makarova	0
+Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?).			Lines 178-9. If PGRMC1 down-regulation is critical, it should be possible to perturb decidualization by expression of an exogenous PGRMC1 protein. This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA. If siRNA-resistant PGRMC1 levels are able to inhibit decidualization it would provide convincing evidence in favour of the hypothesis that lowered PGRMC1 is required for decidualization, and is perhaps even a trigger. A codon-altered PGRMC1 expression vector would cost just several hundred dollars from a provider such as Genscript, so the experiment is eminently achievable.	1	2	reprodmed3020015_makarova	0
+Unfortunately, the quality of experimental design and results processing are somewhat problematic.			Notably, Fig. 4. lacks the western blot equivalent to Fig.3A, showing that siRNA indeed depleted PGRMC1 levels.	1	2	reprodmed3020015_makarova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test. Assuming binomial distribution, a two-way ANOVA is required for these experiments.	1	2	reprodmed3020015_makarova	0
+The latter could well contribute to alterations in PGRMC1 abundance cited by this study.			Line 189-96. The paragraph describes a result showing that the results are independent of P4. However, the control structure of comparisons is incorrect. The simplest direct comparison would be db-cAMP/P4 vs. db-cAMP to permit the argument the authors are presenting. However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4. The statistical significance of any result would be evaluated using multivariate analysis.	1	2	reprodmed3020015_makarova	0
+Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true).			Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1. The results of PMID: 22492871 and PMID: 21109987 MUST be cited here. This is not a novel finding.	1	2	reprodmed3020015_makarova	0
+Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool.			The discussion must qualify the results of Salsano et al. who used AG-205 to inhibit PGRMC1. There was no control in PGRMC1 genomic/CRISPR KO cells (not si/miRNA KO) to demonstrate that effects induced by AG-205 were due to PGRMC1.	1	2	reprodmed3020015_makarova	0
+The authors go on to show that expogenous expression of miR-98 lowers PGRMC1 abundance levels (a more apt description than the fuzzy description of “PGRMC1 downregulation”, which could be by a number of mechanisms as discussed below).			Line 261. mechanisms HAVE not (plural mechanisms)	1	2	reprodmed3020015_makarova	0
+The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation.			Lines 261-262. “In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded. Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) >>	1	2	reprodmed3020015_makarova	0
+Line 261. mechanisms HAVE not (plural mechanisms) Lines 261-262.			Lines 271-76. This is very long and burdens the reader. Always make new sentences when possible. Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy. Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation. Furthermore, controlling their expression may support the improvement of IVF (NOT IFV) outcome. Define IVF at first use.	1	2	reprodmed3020015_makarova	0
+In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			Lines 281-5. PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988). That could be discussed here in terms of mechanism.	1	2	reprodmed3020015_makarova	0
+Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented.			Lines 286-287. Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?). Suggest observations that may be able to discriminate between different possibilities.	1	2	reprodmed3020015_makarova	0
+The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required.			Furthermore, the required experiment is technically not challenging. The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed. The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required. This may be only correlation.	3	2	reprodmed3020015_makarova	0
+The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient.			Lines 142-3. “Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments. All reported results were observed in at least two independent experiments.	3	2	reprodmed3020015_makarova	0
+The revised manuscript as presently resubmitted is a minor revision.			Line 259. PGRMC1-independe (add t) >>	3	2	reprodmed3020015_makarova	0
+The western blot (also Fig.3) result must show the entire molecular weight range (e.g.			Lines 263-267: “Because, the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence. What is being said?	3	2	reprodmed3020015_makarova	0
+Therefore, a resubmission after major revision is recommended.			lines 281-282. There are two natural sentences here. However, as described above AG-205 is not a specific inhibitor of PGRMC1. [full stop, new sentence] Further study of PGRMC1 knockout on decidualization is needed.	3	2	reprodmed3020015_makarova	0
+The authors have addressed most of my concerns.			The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation. Sabbir’s results (ref 31) imply that this also holds for PGRMC1. The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned. To be clear, protein levels can be mediated by altered rates of transcription, translation, or degradation. This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues. Lines 300-307.	3	2	reprodmed3020015_makarova	0
+demonstrate that PGRMC1 abundance levels are higher in proliferative phase versus secretory phase endometrial patient biopsies (fig.1).			In conclusion, if this published was published in present form then it would invariably be cited as demonstrating that PGRMC1 downregulation by miR-98 causes decidualization. Since that is by no means certain, the study should not be published in its present form. The opportunity should remain open for the authors to convincingly demonstrate the requirement for PGRMC1 down-regulation in decidualization by performing the requested experiments, or others of their devising. However, in the first instance the manuscript cannot be published as it stands, and should therefore be rejected now with the stated option of subsequent resubmission only if new evidence is obtained.	5	2	reprodmed3020015_makarova	0
+All reported results were observed in at least two independent experiments.			The reviewer is not convinced that PGRMC1 level attenuation by miR-98 mechanistically drives attenuation. Cell biology is quite complex, and it remains fully feasible that PGRMC1 levels only correlate with the decidualization process. In the lack of convincing evidence, the manuscript should be rejected.	7	2	reprodmed3020015_makarova	0
+If further mechanism exploration is carried out, the research will be more complete and persuasive.			Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？The sample size is small. And please provide basic information of the patient.	1	2	reprodmed3020015_makarova	0
+The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient.			The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough. It is suggested to add cell lines or normal endometrial cells for verification. If further mechanism exploration is carried out, the research will be more complete and persuasive.	1	2	reprodmed3020015_makarova	0
+If further mechanism exploration is carried out, the research will be more complete and persuasive.			The author provided explanations for the questions. The manuscript has been modified accordingly. I think the manuscript could be accepted now.	3	2	reprodmed3020015_makarova	0
+The authors mention technical difficulties in establishing ESCs that express miRNA-resistant PGRMC1.			As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	reprodmed3020015_makarova	0
+However, as described above AG-205 is not a specific inhibitor of PGRMC1.			1) In general, the manuscript refers often to PGRMC1 expression or downregulation. These are fuzzy terms for proteins. It is much preferable to refer to protein abundance, and changes in abundance. Such changes may occur at the translational or post-translational level. Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool. The authors are strongly advised to change from e.g. ‘expression’ to ‘abundance’ throughout.	1	2	reprodmed3020015_perova	0
+PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491).			2) Additionally, the methods do not say that all results were repeated at least once, and that a representative results figure is presented. Therefore, the reviewer assumes that experiments have only been performed once. The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.	1	2	reprodmed3020015_perova	0
+Furthermore, the required experiment is technically not challenging.			3) Line 24. AG-205. The authors must refer to the fact that AG-205 is not specific for PGRMC1, contrary to many citations in the literature (such as would be the case here, if uncorrected). Cite PMID: 34944026, PMID: 34680104, PMID: 32924377. Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.	1	2	reprodmed3020015_perova	0
+That could be discussed here in terms of mechanism.			4) It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).	1	2	reprodmed3020015_perova	0
+Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.			5) Line 50. “(10-12); however,” Two discrete sentences “(10-12).	1	2	reprodmed3020015_perova	0
+It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.			6) Lines 76-78. Statistical sample sizes were unfavourably small. The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).	1	2	reprodmed3020015_perova	0
+The revised manuscript as presently resubmitted is a minor revision.			7) Lines 88-98. Cell culture. What characterization was performed to confirm the identity of cells as ESCs? In general, there is very little detail on these primary cultured lines. How long are they held in culture? Are they stored cryogenically? How are they re-seeded into culture?	1	2	reprodmed3020015_perova	0
+A codon-altered PGRMC1 expression vector would cost just several hundred dollars from a provider such as Genscript, so the experiment is eminently achievable.			8) Lines 121-133. What percentage acrylamide were the SDS PAGE gels. For what length of time and at what voltage settings was electrophoretic transfer to PVDF? In what device?	1	2	reprodmed3020015_perova	0
+The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			9) Line 126-7. Give the catalogue number of the secondary antibody.	1	2	reprodmed3020015_perova	0
+Figure 2 seems to depict just one cell culture (which is not even named), with 4 replicates of each measurement.			10) Line 131. On what device was ECL light detected? What were the device settings? How was pixel saturation dealt with?	1	2	reprodmed3020015_perova	0
+In general, there is very little detail on these primary cultured lines.			11) Line 13 Which ImageJ plug-in was used for quantification?	1	2	reprodmed3020015_perova	0
+Our responses to your comments are as follows.			12) Lines 134-137. Result errors should not be expressed as SEM, but as standard deviation (s.d.). All results should show s.d. error bars.	1	2	reprodmed3020015_perova	0
+And please provide basic information of the patient.			13) Line 150. The legend to fig. 1. inappropriately ends with “2.2. Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2.	1	2	reprodmed3020015_perova	0
+There is some concern about the specificity of the AG-205 inhibitor here and throughout, which weakens the deductive reasoning and is discussed below.			14) Lines 153-156, Figure 2. The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient. This result must be demonstrated in a substantively convincing manner.	1	2	reprodmed3020015_perova	0
+At the very least this mechanism merits detailed discussion in that section.			2. 15) Line 158/Fig. The western blot (also Fig.3) result must show the entire molecular weight range (e.g. in supplemental data), including MW markers. PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491). The latter could well contribute to alterations in PGRMC1 abundance cited by this study. It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.	1	2	reprodmed3020015_perova	0
+However, in the first instance the manuscript cannot be published as it stands, and should therefore be rejected now with the stated option of subsequent resubmission only if new evidence is obtained.			16) Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true).	1	2	reprodmed3020015_perova	0
+Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.			17) Line 163. Change 3.2. to 3.3.	1	2	reprodmed3020015_perova	0
+Figure 2 seems to depict just one cell culture (which is not even named), with 4 replicates of each measurement.			18) Lines 164-172. This cell culture experiment also seems to be based upon just one primary cell culture, split into replicates. I am uncertain because the methods do not explain how the experiment was performed. Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.	1	2	reprodmed3020015_perova	0
+In primary endometrial stem cell (ESC) culture they demonstrate that chemical induction (by db-cAMP/P4) of pseudo-decidualization in culture is accompanied by decreased PGRMC1 protein levels (fig.2).			19) Lines 178-9. If PGRMC1 down-regulation is critical, it should be possible to perturb decidualization by expression of an exogenous PGRMC1 protein. This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA. If siRNA-resistant PGRMC1 levels are able to inhibit decidualization it would provide convincing evidence in favour of the hypothesis that lowered PGRMC1 is required for decidualization, and is perhaps even a trigger. A codon-altered PGRMC1 expression vector would cost just several hundred dollars from a provider such as Genscript, so the experiment is eminently achievable.	1	2	reprodmed3020015_perova	0
+“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments.			20) Notably, Fig. 4. lacks the western blot equivalent to Fig.3A, showing that siRNA indeed depleted PGRMC1 levels.	1	2	reprodmed3020015_perova	0
+Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?).			21) Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test. Assuming binomial distribution, a two-way ANOVA is required for these experiments.	1	2	reprodmed3020015_perova	0
+Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？ The sample size is small.			22) Line 189-96. The paragraph describes a result showing that the results are independent of P4. However, the control structure of comparisons is incorrect. The simplest direct comparison would be db-cAMP/P4 vs. db-cAMP to permit the argument the authors are presenting. However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4. The statistical significance of any result would be evaluated using multivariate analysis.	1	2	reprodmed3020015_perova	0
+The western blot (also Fig.3) result must show the entire molecular weight range (e.g.			23) Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1. The results of PMID: 22492871 and PMID: 21109987 MUST be cited here. This is not a novel finding.	1	2	reprodmed3020015_perova	0
+Author Response Responses to Reviewer 2 We appreciate your review of our manuscript and constructive comments and suggestions.			24) The discussion must qualify the results of Salsano et al. who used AG-205 to inhibit PGRMC1. There was no control in PGRMC1 genomic/CRISPR KO cells (not si/miRNA KO) to demonstrate that effects induced by AG-205 were due to PGRMC1.	1	2	reprodmed3020015_perova	0
+[full stop, new sentence] Further study of PGRMC1 knockout on decidualization is needed.			25) Line 261. mechanisms HAVE not (plural mechanisms) >>	1	2	reprodmed3020015_perova	0
+It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.			26) Lines 261-262. “In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded. Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) >>	1	2	reprodmed3020015_perova	0
+Therefore, a resubmission after major revision is recommended.			27) Lines 271-76. This is very long and burdens the reader. Always make new sentences when possible. Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy. Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation. Furthermore, controlling their expression may support the improvement of IVF (NOT IFV) outcome. Define IVF at first use.	1	2	reprodmed3020015_perova	0
+If higher PGRMC1 levels are indeed critical for the suppression of decidualization, as is reasoned by the authors, then it should be possible to maintain PGRMC1 levels artificially to inhibit decidualization, as discussed below.			28) Lines 281-5. PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988). That could be discussed here in terms of mechanism.	1	2	reprodmed3020015_perova	0
+If further mechanism exploration is carried out, the research will be more complete and persuasive.			29) Lines 286-287. Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?). Suggest observations that may be able to discriminate between different possibilities.	1	2	reprodmed3020015_perova	0
+The authors go on to show that expogenous expression of miR-98 lowers PGRMC1 abundance levels (a more apt description than the fuzzy description of “PGRMC1 downregulation”, which could be by a number of mechanisms as discussed below).			In response the authors have made minimal changes to the manuscript, effectively restricted to rewording the title, and several minor text changes. Unfortunately, the authors have been unable to demonstrate the direct involvement of PGRMC1 downregulation in decidualization. Since this is the main publishable finding, the results as they stand do not merit publication. The authors mention technical difficulties in establishing ESCs that express miRNA-resistant PGRMC1. This may be due to stable plasmid selection conditions, which may affect the differentiation status of the ESC cells. It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1. An inducible vector may be advisable, such as tetracycline, IPTG, or other available systems. Such vectors typically include fluorescent protein reporters, enabling discrimination between infected and non-infected cells in one plate, or population separation by FACS.	3	2	reprodmed3020015_perova	0
+Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			The inclusion of the model of Fig. 6 in the manuscript depicts the problem. This manuscript will be cited as evidence of that inconclusive model. The reviewer is not convinced that PGRMC1 level attenuation by miR-98 mechanistically drives attenuation. Cell biology is quite complex, and it remains fully feasible that PGRMC1 levels only correlate with the decidualization process. In the lack of convincing evidence, the manuscript should be rejected.	5	2	reprodmed3020015_perova	0
+PGRMC1 protein instability could be the dominant effect involved, with miRNA transcript regulation playing only a minor part.			Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？	1	2	reprodmed3020015_perova	0
+Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2.			The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough. It is suggested to add cell lines or normal endometrial cells for verification. If further mechanism exploration is carried out, the research will be more complete and persuasive.	1	2	reprodmed3020015_perova	0
+Furthermore, controlling their expression may support the improvement of IVF (NOT IFV) outcome.			I requested a major revision including the use of miRNA-resistant exogenous PGRMC1 expression (altered codon usage) to demonstrate that lack of PGRMC1 attenuation will prevent decidualization. That new experimental result is what elevated the required changes to a major revision. The revised manuscript as presently resubmitted is a minor revision. This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization. This requires stronger evidence than that currently presented. Furthermore, the required experiment is technically not challenging. The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed. The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required. This may be only correlation. Before such a claim is published in the scientific literature, it must be supported by convincing evidence. Aspiration to clarify this in future papers does not merit publication of the present paper.	1	2	reprodmed3020015_perova	0
+Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.			Lines 142-3.“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments. All reported results were observed in at least two independent experiments.	1	2	reprodmed3020015_perova	0
+Author Response We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript.			Line 259. PGRMC1-independe (add t) >>	1	2	reprodmed3020015_perova	0
+Suggest observations that may be able to discriminate between different possibilities.			Lines 263-267: “Because the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence.	1	2	reprodmed3020015_perova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualisation.			There are two natural sentences here. However, as described above AG-205 is not a specific inhibitor of PGRMC1. [full stop, new sentence] Further study of PGRMC1 knockout on decidualization is needed.	1	2	reprodmed3020015_perova	0
+Author Response Responses to Reviewer 2 We appreciate your review of our manuscript and constructive comments and suggestions.			The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation. Sabbir’s results (ref 31) imply that this also holds for PGRMC1. The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned. To be clear, protein levels can be mediated by altered rates of transcription, translation, or degradation. This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues.	1	2	reprodmed3020015_perova	0
+Point 5: What is the main difference (except of the terrestrial data over China) to other combined GFMs, such as XGM2016 or XGM2019?			What is the main methodological difference and advantage of your method? Please comment on that question quantitatifely and qualitatively.	1	2	rs14061437_makarova	0
+The point-by-point response can be found from attachment, please see attachment.			Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach? Could you please comment on the effects you introduce by applying such a sharp truncation combination approach.	1	2	rs14061437_makarova	0
+If you have any information, please don’t hesitate to let us know.			Page 2, Line 60. Change “,” to “.”	1	2	rs14061437_makarova	0
+It has improved the readability, clarity, and quality of our manuscript.			Page 2, Line 62. Change “to use of high-accuracy” to “to use high-accuracy” Thank you.	1	2	rs14061437_makarova	0
+It has improved the readability, clarity, and quality of our manuscript.			Page 11, Line 356. Change “not reasonable” to “not a reasonable” Thank you.	1	2	rs14061437_makarova	0
+If you have any information, please don’t hesitate to let us know.			Page 16, Line 488. Considering the STD you should rather not give the second digit after the comma.	1	2	rs14061437_makarova	0
+43 This is due to the fact that the MSL presents geographical and time-dependent variations, 44 Citation: Lastname, F.; Lastname, F.; Lastname, F. Title.			Generally it is a good paper and it was interesting to me. However, comparing with the conclusions (I mean, the computed potential value) the paper is a bit too long. I can accept it, because it is a good explanatory text, however some partially offtopic or superfluous parts (e.g. point 2.1.2 (keeping the Table 1 only), the unified topo data in lines 181-191, the „textbook equations” of the gravity of a prism in page 7 and the textbook adjustment equations in page 8). It is rather an editorial question. The paper is good and acceptable without them, too.	1	2	rs14061437_makarova	0
+The point-by-point response can be found from attachment, please see attachment.			Line 259 refers to a x as a ’parameter’ in Eq 12, I suggest to use ’vector’ or ’parameter vector’ instead.	1	2	rs14061437_makarova	0
+Response to Reviewer 2 Comments: We thank you for your constructive and detailed comments.			As a structural suggestion, considerable part of the Results are rather belong to Discussion (thus making the better balance between these two chapters, in extent and in content, too): I suggest point 3.3 (or maybe also 3.2) to Discussion – as they are discussing the already shown results. Conclusion – it shouldn’t be a summary as should be recompiled. The second part of the 3rd paragraph and the 4th paragraph are enough, completed by the estimated error of the main conclusive potantial value.	1	2	rs14061437_makarova	0
+The estimation of height datum geopotential value usually depends on high- 15 precision global gravity field models (GFMs).			Page 2, Line 60. Change “,” to “.”	1	2	rs14061437_perova	0
+Your method for combining the satellte-only GFM with EGM2008 (sec.			Page 2, Line 62. Change “to use of high-accuracy” to “to use high-accuracy” Thank you.	1	2	rs14061437_perova	0
+Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach?			Page 11, Line 356. Change “not reasonable” to “not a reasonable” Thank you.	1	2	rs14061437_perova	0
+Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach?			Page 16, Line 488. Considering the STD you should rather not give the second digit after the comma.	1	2	rs14061437_perova	0
+2022, 14, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Last- name Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations.			What is the main difference (except of the terrestrial data over China) to other combined GFMs, such as XGM2016 or XGM2019? What is the main methodological difference and advantage of your method? Please comment on that question quantitatifely and qualitatively.	1	2	rs14061437_perova	0
+We have added descriptions and contents in the revised manuscript.			3.3) seems quite simple for me and probably not optimal. Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach? Could you please comment on the effects you introduce by applying such a sharp truncation combination approach. Your method for combining the satellite-only GFM with EGM2008 (sec.	1	2	rs14061437_perova	0
+If you have any information, please don’t hesitate to let us know.			Generally it is a good paper and it was interesting to me. However, comparing with the conclusions (I mean, the computed potential value) the paper is a bit too long. I can accept it, because it is a good explanatory text, however some partially offtopic or superfluous parts (e.g. point 2.1.2 (keeping the Table 1 only), the unified topo data in lines 181-191, the „textbook equations” of the gravity of a prism in page 7 and the textbook adjustment equations in page 8). It is rather an editorial question. The paper is good and acceptable without them, too.	1	2	rs14061437_perova	0
+The height related to the Earth's 37 gravity field usually refers to the orthometric or normal height, which is strictly based on 38 the geopotential number CP ( 0 LVD P P C W W   , where 0 LVD W is the geopotential value for 39 the local vertical datum, and WP indicates the gravity potential for point P).			Line 259 refers to a x as a ’parameter’ in Eq 12, I suggest to use ’vector’ or ’parameter vector’ instead.	1	2	rs14061437_perova	0
+To compen- 19 sate for the omission errors in satellite-only GFMs, a spectral expansion approach is used to obtain 20 the refined gravity field models using the EGM2008 (Earth Gravitational Model 2008) and residual 21 terrain model (RTM) technique.			As a structural suggestion, considerable part of the Results are rather belong to Discussion (thus making the better balance between these two chapters, in extent and in content, too): I suggest point 3.3 (or maybe also 3.2) to Discussion – as they are discussing the already shown results. Conclusion – it shouldn’t be a summary as should be recompiled. The second part of the 3rd paragraph and the 4th paragraph are enough, completed by the estimated error of the main conclusive potantial value.	1	2	rs14061437_perova	0
+1 Response to Reviewer 1 Comments Comments 1 Please add a table with patients' baseline characteristics, including BMI and smoking status.			Please add a table with patients' baseline characteristics, including BMI and smoking status.	1	2	s22072682_makarova	0
+Comments 2 - Given the poor quality of the images (Fig 1 and 2), it might be better to redraw them and describe them as an adaptation of the original publication.			Did authors estimate sample size? Please add in data analysis section	1	2	s22072682_makarova	0
+Response 3: I described baseline load of the participants.			Please add ethics protocol approval number	1	2	s22072682_makarova	0
+1 Response to Reviewer 1 Comments Comments 1 Please add a table with patients' baseline characteristics, including BMI and smoking status.			I found some minor English errors throughout the paper. Please have a re-check.	1	2	s22072682_makarova	0
+Comments 4 - In figure 3, the indication of the test time appears in the reverse direction.			I suggest presenting the paper's aim as the last idea in the introduction to make the manuscript easier to read.	1	2	s22072682_makarova	0
+Comments 4 I found some minor English errors throughout the paper.			Given the poor quality of the images (Fig 1 and 2), it might be better to redraw them and describe them as an adaptation of the original publication.	1	2	s22072682_makarova	0
+Comments 4 I found some minor English errors throughout the paper.			The methods do not specify how the baseline load of the participants was determined.	1	2	s22072682_makarova	0
+Comments 4 I found some minor English errors throughout the paper.			"In figure 3, the indication of the test time appears in the reverse direction. Time ""0"" should be at the start."	1	2	s22072682_makarova	0
+Comments 5 - I recommend deleting graphs 5, 8, and 11 because graphs 7, 10, and 13 show the same information together with lines obtained by analyzing the structural change of the time series.			I recommend deleting graphs 5, 8, and 11 because graphs 7, 10, and 13 show the same information together with lines obtained by analyzing the structural change of the time series.	1	2	s22072682_makarova	0
+Especially since “there does not appear to be any literature focused on examining the association between the constitution of remuneration committees and the reporting of CSR information”, the authors should provide the theoretical link to propose the hypotheses.			Theoretical issues: although the agency theory and practical function of remuneration committees were described in the manuscript, few efforts were made to give the main theoretical mechanism/logic. Especially since “there does not appear to be any literature focused on examining the association between the constitution of remuneration committees and the reporting of CSR information”, the authors should provide the theoretical link to propose the hypotheses.	1	2	su14020860_makarova	0
+Author Response Dear Reviewer 1, We are sending you the second version of our paper.			Methodological issues: the dummy of CSR_index is too simple and cannot address the complexity of CSR strategy and CSR disclosure. And so is the independent variable. Many other important control variables should be introduced to the regression, such as, financial performance, the firm size, the firm age, the industry-fixed effects, the country-fixed effects. The results may not be robust without these controls.	1	2	su14020860_makarova	0
+Now, we would firstly like to thank you for your efforts in revising our study and we hope that the arguments and changes are satisfactory.			Other issues: the figure 1 can be re-designed to conform the academic norms.	1	2	su14020860_makarova	0
+We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			The article is methodologically OK and well written. However, in general, we see many articles like this in the area of social and environmental reporting. Also, the application of agency theory is so common that reading it over and again in the new studies also makes the readers bored these days. Now it is important that the new researchers try to take the research on this area to a new and interesting level. However, I accept this article only for the moderating variable the authors have tested.	1	2	su14020860_makarova	0
+We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			The introduction lacks of motivation. The reader needs to understand why your research question is important.	1	2	su14020860_makarova	0
+Author Response Dear Reviewer 3, Thank you for your kind comments.			The theoretical background is weak. I am not convinced on your explanation on why committees would affect CSR.	1	2	su14020860_makarova	0
+The reader needs to understand why your research question is important.			Your empirical model needs to be reconsidered. Indeed, I believe you have some omitted control variables which may cause an endogeneity issue and drive your findings	1	2	su14020860_makarova	0
+Best regards, The authors Author Response File: Author Response.docx			The paper lacks of robustness checks	1	2	su14020860_makarova	0
+We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			The conclusion needs to be rewritten to highlight the practical contributions of the paper	1	2	su14020860_makarova	0
+We are hopeful you find our responses/edits to your concerns (expressed below) sufficient.			although the agency theory and practical function of remuneration committees were described in the manuscript, few efforts were made to give the main theoretical mechanism/logic. Especially since “there does not appear to be any literature focused on examining the association between the constitution of remuneration committees and the reporting of CSR information”, the authors should provide the theoretical link to propose the hypotheses.	1	2	su14020860_perova	0
+Now it is important that the new researchers try to take the research on this area to a new and interesting level.			the dummy of CSR_index is too simple and cannot address the complexity of CSR strategy and CSR disclosure. And so is the independent variable. Many other important control variables should be introduced to the regression, such as, financial performance, the firm size, the firm age, the industry-fixed effects, the country-fixed effects. The results may not be robust without these controls.	1	2	su14020860_perova	0
+We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			the figure 1 can be re-designed to conform the academic norms.	1	2	su14020860_perova	0
+Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			The article is methodologically OK and well written. However, in general, we see many articles like this in the area of social and environmental reporting. Also, the application of agency theory is so common that reading it over and again in the new studies also makes the readers bored these days. Now it is important that the new researchers try to take the research on this area to a new and interesting level. However, I accept this article only for the moderating variable the authors have tested.	1	2	su14020860_perova	0
+We had the firm size variable labelled as LTA which is calculated as logarithm of total assets.			The paper studies the relationship between remuneration committees and CSR as well as the moderating role of independent directors. While the paper seems interesting I have several concerns that I would like to share with the authors: The introduction lacks of motivation. The reader needs to understand why your research question is important.	1	2	su14020860_perova	0
+Please, notice that our responses are provided in bold after your comments/suggestions.			The theoretical background is weak. I am not convinced on your explanation on why committees would affect CSR.	1	2	su14020860_perova	0
+Please, notice that our responses are provided in bold after your comments/suggestions.			Your empirical model needs to be reconsidered. Indeed, I believe you have some omitted control variables which may cause an endogeneity issue and drive your findings	1	2	su14020860_perova	0
+The reader needs to understand why your research question is important.			The paper lacks of robustness checks	1	2	su14020860_perova	0
+Please, notice that our responses are provided in bold after your comments/suggestions.			The conclusion needs to be rewritten to highlight the practical contributions of the paper	1	2	su14020860_perova	0
+It is also essential to make a better connection between these notions, since the relationship between them is not evident per se.” Governance approaches mentioned are now related to Transformative Social-Ecological Innovation (TSEI), which are innovations with explicit transformative goals and attention to mutual gains, and originating from political science and negotiation theory in particular.			This paper seeks to develop the notion of a Natural Social Contract (NSC) to enable transformative governance based on co-evolution. In a nutshell, to quote, the NSC “involves the fine-tuning of top-down policy and visions with important bottom-up processes (…), and takes place in a hybrid sphere, in which there is room for collaboration between governments, businesses, knowledge institutions and civil society, among others, characterized by Penta-helix models and based on multiple value creation”. Because of this collaborative, polycentric approach, the papers draw on Evolutionary Governance Theory, and develops a perspective of “co-evolutionary steering of interrelated transitions”. The paper’s aspiration is clear and laudable; yet its elaboration is wanting. My recommendation therefore leans towards a strong ‘revise and resubmit’, based on these five points.	1	2	su14052976_makarova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			First, my main problem is that the paper reads as a compilation of manifestos, rather than one argumentative thread. From begin to end, the paper presents a mix of literature review and substantive argumentation, plus some examples. My recommendation is to set out a clearer agenda, in which the first part delves into the literature to explain the key issue, key concepts and key dilemmas, and a second part presents solutions and directions meeting the key questions and dilemmas. In brief, the paper warrants more focus, edge and structure.	1	2	su14052976_makarova	0
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			A second point is the limited elaboration of the social contract, including the natural aspect. A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement. Fig2. moreover, details many aspects. What remains unclear and implicit, however, is the form and operationalisation of such contract. How is the social contract articulated; how is consent manifested? How does one conceive and apply natural ‘design principles’ (669)? This also involves a question of scale: how do agreements at project level chime with societal notions of Social Contracts? How does a project mission contribute to a broader social contract? Much of the joining forces in the hybrid sphere between institutional domains may be primarily instrumental (pooling resources, aligning interests, preventing holdups), below the scope of an (aspired) social contract. What kind of hybrid practices gave shape to the latter? And how then is the aspect of nature taken on board?	1	2	su14052976_makarova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			A third point concerns the link between theory and practice. The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary. Much of this, moreover, is action-oriented, pointing out major carriers and direction of change. Two cases serve as quick illustrations, loosely underwriting some key statements. This presents, in my view, too much of a gap. The paper could do more to translate the general aspects of transformative governance into more specific project aspects, as an evaluative framework for debating the cases.	1	2	su14052976_makarova	0
+What could be helpful is, in your argumentation, briefly incorporate the genealogy of your key concepts, while you work with them, thus showing more clearly how you combine and transform them.			Fourth, the section on co-evolution warrants serious re-thinking. Rather than elaborating the notion and development of NSC, this section largely discusses co-evolutionary approaches, with emphasis on discursive aspects (e.g., metaphors, open concepts and master signifiers). While the section aims to uncover the generative mechanisms behind a ‘transition to a Natural Social Contract’, it lacks focus and thread. In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI). This raises two basic problems. One, the definition and position of NSC in this conceptual medley remains vague. Two, the nature and form of evolutionary variation, selection and dynamics remain unclear. Co-evolution is now associated with all kinds of change-in-tandem, social-sustainable, different institutional actors, discursive items, etc. One wonders whether the notion of transformative change really warrant this evolutionary perspective.	1	2	su14052976_makarova	0
+"The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the?"			Following on the latter, my final point concerns the relation between transformative change and NSC. One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc. On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118). I would suggest the paper can delve deeper into the question of how the NSC can help to meet and overcome such resistances, and to bring forces of change and resistance more in line in the story.	1	2	su14052976_makarova	0
+20 Reviewer 3: The paper presents quite an interesting discussion.			Source citations are sometimes unclear, as they refer to “	1	2	su14052976_makarova	0
+Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.			"Language is generally okay. The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the? coevolution"", ""health care"", ""es"" etc....)"	3	2	su14052976_makarova	0
+This paper has much improved, also because its scope is clearer.			However, the paper in many parts takes an extreme normative tone that weakens its scientific contribution. It is important that the authors give more emphasis on the scientific evidences of what they defend and anchor their argumentation in such evidences, whether theoretical or empirical.	1	2	su14052976_makarova	0
+Figure 2: Possible systemic leverage points for a societal transformation towards a Natural Social Contract, through transformative governance based on co-evolution across several interrelated dimensions (this figure is a synthesis of table 3.4 in Huntjens, 2021) Figure 3: The Intervention Flower as a tool for identifying options for co-evolutionary steering through connecting actor-coalitions and interdependent systemic leverage points.			Make the problematization clearer, exploring what motivates the proposed theoretical study and in which previous studies it is anchored. In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.	1	2	su14052976_makarova	0
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			"Remove from the problematization the ""normative-prescriptive"" tone starting from questions and nuisances that justify the study and from gaps in the literature. I am referring to universal and generalist statements with a ""should be"" tone that do not fit well with a scientific article, like the one in lines 51 to 54, among many others in the article."	1	2	su14052976_makarova	0
+2 Editor: After receiving the three reviews, we would still recommend major revision.			Make clearer and more explicit how the article meets the proposed problematic and responds to the gaps found in the debate, including raising the limits of such proposition.	1	2	su14052976_makarova	0
+Much of this, moreover, is action-oriented, pointing out major carriers and direction of change.			"Further problematize the discussions/debates that are being mobilized in the article. The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial. It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates. To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy. Only in these aspects it is possible to bring several elements that problematize the discussion and show that there is not a single path or model for building public governance that favors sustainability."	1	2	su14052976_makarova	0
+It is important that the authors give more emphasis on the scientific evidences of what they defend and anchor their argumentation in such evidences, whether theoretical or empirical.			In terms of the debate on public governance it would be important to insert the discussion of co-evolutionary governance not only ideally, but also exploring the differences between this theoretical proposition and other forms of governance already discussed in the literature as collaborative governance (Ansell and Gash, 2015), for example. Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.	1	2	su14052976_makarova	0
+Cover letter to Journal ‘Sustainability’ Special Issue: Environmental Policy and Governance: Evolutionary Perspectives Subject: Point by point response to editor’s and reviewers’ comments Date: 1 February 2022 Manuscript ID: sustainability-1512113 Manuscript title: Transition policy after corona: The importance of a Natural Social Contract and co-evolutionary governance No.			"Empirical cases are very relevant, but are placed as ""mere examples"" without delving into the challenges and difficulties lived in practice. In this sense, it is not clear the relationship between the propositions presented by the authors in section 4 and the empirical or even theoretical evidences presented in the article. I suggest to improve the interface between the propositions presented and what comes before in the text."	1	2	su14052976_makarova	0
+One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive.			The authors made a visible effort of revision, introducing new sections and discussing better the debates on which their arguments are based. I don't see a problem in a theoretical article to be normative, however some problems in the paper remain and it still need a revision to be published. Specially it needs better clarity in terms of focus and structure. Here are my recommendations:”	3	2	su14052976_makarova	0
+This would take care of the remarks about case studies and examples, and avoid the impression that there's several manifestos, or a manifesto immediately derived from one empirical situation.			The text puts in relation 03 new notions that need to be better related by the authors: evolutionary/transformative governance, transformative socioecological innovation and natural social contract. These notions are proposed by the authors and based on their previous work (since they rely on self-citation). Therefore, it seems essential to me to make it clear in each of the sections of the article that deal with these notions from where they start and how they relate to previous debates to support the authors' arguments. It is also essential to make a better connection between these notions, since the relationship between them is not evident per se.”	3	2	su14052976_makarova	0
+8 Reviewer 1: A second point is the limited elaboration of the social contract, including the natural aspect.			"In this sense, I recommend to start with the broader discussion of the natural contract (beggining of session 3), ending with the question of how to put in practice this social contract, what challenges (what is the main focus of this text). The line 160 present this question as: ""The big question for steering is: how to initiate and accelerate system changes whose features are not perfectly clear and often times underdeveloped compared to the existing systems? This would make room to bring session 02 that addresses evolutionary/transformative governance to the TSEI.”"	3	2	su14052976_makarova	0
+We have included a reference to the anxiety and (absent) feelings of control and competence.			"The section 02 ""Transformation pleas and approach to achieve this"" can be improved. I recommend reviewing the title referring to the governance debate that is addressed in the section, for example: ""Transformative pleas and approaches of governance to achieve this""...It is also important to better relate/ differentiate the evolutionary governance proposed by authors and the others discussed in the literature (adaptive, reflexive, deliberative, collaborative, etc.). The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution. I suggest that the authors present the debate and the different definitions and then present the notion of GE, relating it to the TSEI.”"	3	2	su14052976_makarova	0
+Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.			The examples placed in section 03 could compose a section in itself, which sought to relate theory and practical examples (not cases), illustrating the proposed model. It will be nice to put forward the discussion of the conceptual variables of transformative governance proposed.	3	2	su14052976_makarova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			It remains not clear the relationship between the propositions presented by the authors in section 4 and the discutions in other sections. I suggest to improve the interface between the propositions presented and what comes before in the text, specialy linking with the contributions and limites of the proposal to face this challenges.	3	2	su14052976_makarova	0
+2 Editor: After receiving the three reviews, we would still recommend major revision.			Formal aspects: The text needs a language revision and formatting of the figures to make them more readable. Avoid the excessive use of acronyms and review them throughout the text. I thank the authors and the editor for this opportunity to review and discuss this interesting paper and I hope that my comments help to improve the paper. Best regards.	3	2	su14052976_makarova	0
+Figure 2: Possible systemic leverage points for a societal transformation towards a Natural Social Contract, through transformative governance based on co-evolution across several interrelated dimensions (this figure is a synthesis of table 3.4 in Huntjens, 2021) Figure 3: The Intervention Flower as a tool for identifying options for co-evolutionary steering through connecting actor-coalitions and interdependent systemic leverage points.			I'll suggest a format review in the figures and their quotations, as well as the box and the table.	1	2	su14052976_makarova	0
+20 Reviewer 3: The paper presents quite an interesting discussion.			First, my main problem is that the paper reads as a compilation of manifestos, rather than one argumentative thread. From begin to end, the paper presents a mix of literature review and substantive argumentation, plus some examples. My recommendation is to set out a clearer agenda, in which the first part delves into the literature to explain the key issue, key concepts and key dilemmas, and a second part presents solutions and directions meeting the key questions and dilemmas. In brief, the paper warrants more focus, edge and structure.	1	2	su14052976_perova	0
+"19 Reviewer 2: 6) Empirical cases are very relevant, but are placed as ""mere examples"" without delving into the challenges and difficulties lived in practice."			A second point is the limited elaboration of the social contract, including the natural aspect. A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement. Fig2. moreover, details many aspects. What remains unclear and implicit, however, is the form and operationalisation of such contract. How is the social contract articulated; how is consent manifested? How does one conceive and apply natural ‘design principles’ (669)? This also involves a question of scale: how do agreements at project level chime with societal notions of Social Contracts? How does a project mission contribute to a broader social contract? Much of the joining forces in the hybrid sphere between institutional domains may be primarily instrumental (pooling resources, aligning interests, preventing holdups), below the scope of an (aspired) social contract. What kind of hybrid practices gave shape to the latter? And how then is the aspect of nature taken on board?	1	2	su14052976_perova	0
+It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates.			A third point concerns the link between theory and practice. The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary. Much of this, moreover, is action-oriented, pointing out major carriers and direction of change. Two cases serve as quick illustrations, loosely underwriting some key statements. This presents, in my view, too much of a gap. The paper could do more to translate the general aspects of transformative governance into more specific project aspects, as an evaluative framework for debating the cases.	1	2	su14052976_perova	0
+While the section aims to uncover the generative mechanisms behind a ‘transition to a Natural Social Contract’, it lacks focus and thread.			Fourth, the section on co-evolution warrants serious re-thinking. Rather than elaborating the notion and development of NSC, this section largely discusses co-evolutionary approaches, with emphasis on discursive aspects (e.g., metaphors, open concepts and master signifiers). While the section aims to uncover the generative mechanisms behind a ‘transition to a Natural Social Contract’, it lacks focus and thread. In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI). This raises two basic problems. One, the definition and position of NSC in this conceptual medley remains vague. Two, the nature and form of evolutionary variation, selection and dynamics remain unclear. Co-evolution is now associated with all kinds of change-in-tandem, social-sustainable, different institutional actors, discursive items, etc. One wonders whether the notion of transformative change really warrant this evolutionary perspective.	1	2	su14052976_perova	0
+The case at local and regional levels are well developed, but the discussion at international level seems poor, maybe the authors could exclude this mentioned focus o the Introduction I'll suggest a format review in the figures and their quotations, as well as the box and the table.			Following on the latter, my final point concerns the relation between transformative change and NSC. One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc. On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118). I would suggest the paper can delve deeper into the question of how the NSC can help to meet and overcome such resistances, and to bring forces of change and resistance more in line in the story.	1	2	su14052976_perova	0
+!” (France in anger) was a popular facebook site.			Source citations are sometimes unclear, as they refer to “	1	2	su14052976_perova	0
+The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution.			"This paper has much improved, also because its scope is clearer. I am still puzzled about the selection aspect of EGT, notably the specific roles of metaphors, open concepts and master signifiers play here (as objects and agents of selection) - particularly now NCS has been presented as a master signifier. But that may be for a later argument. Language is generally okay. The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the? coevolution"", ""health care"", ""es"" etc....)”"	3	2	su14052976_perova	0
+Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.			1) Make the problematization clearer, exploring what motivates the proposed theoretical study and in which previous studies it is anchored. In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.	1	2	su14052976_perova	0
+It will be nice to put forward the discussion of the conceptual variables of transformative governance proposed.			"2) ""Remove from the problematization the """"normative-prescriptive"""" tone starting from questions and nuisances that justify the study and from gaps in the literature."" I am referring to universal and generalist statements with a ""should be"" tone that do not fit well with a scientific article, like the one in lines 51 to 54, among many others in the article."	1	2	su14052976_perova	0
+The line of argumentation has been greatly improved (something we are happy about ourselves).			3) Make clearer and more explicit how the article meets the proposed problematic and responds to the gaps found in the debate, including raising the limits of such proposition.	1	2	su14052976_perova	0
+What kind of hybrid practices gave shape to the latter?			"4) Further problematize the discussions/debates that are being mobilized in the article. The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial. It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates. To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy. Only in these aspects it is possible to bring several elements that problematize the discussion and show that there is not a single path or model for building public governance that favors sustainability."	1	2	su14052976_perova	0
+How does one conceive and apply natural ‘design principles’ (669)?			5) In terms of the debate on public governance it would be important to insert the discussion of co-evolutionary governance not only ideally, but also exploring the differences between this theoretical proposition and other forms of governance already discussed in the literature as collaborative governance (Ansell and Gash, 2015), for example. Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.	1	2	su14052976_perova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			"6) ""Empirical cases are very relevant, but are placed as """"mere examples"""" without delving into the challenges and difficulties lived in practice."" In this sense, it is not clear the relationship between the propositions presented by the authors in section 4 and the empirical or even theoretical evidences presented in the article. I suggest to improve the interface between the propositions presented and what comes before in the text."	1	2	su14052976_perova	0
+In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.			We want to sincerely thank you for your interest in the Sustannability and for sharing your work with us. I hope this review will encourage your work and the improvement of the text.	1	2	su14052976_perova	0
+In a nutshell, to quote, the NSC “involves the fine-tuning of top-down policy and visions with important bottom-up processes (…), and takes place in a hybrid sphere, in which there is room for collaboration between governments, businesses, knowledge institutions and civil society, among others, characterized by Penta-helix models and based on multiple value creation”.			1) The text puts in relation 03 new notions that need to be better related by the authors: evolutionary/transformative governance, transformative socioecological innovation and natural social contract. These notions are proposed by the authors and based on their previous work (since they rely on self-citation). Therefore, it seems essential to me to make it clear in each of the sections of the article that deal with these notions from where they start and how they relate to previous debates to support the authors' arguments. It is also essential to make a better connection between these notions, since the relationship between them is not evident per se.”	3	2	su14052976_perova	0
+For more information see section 2 on page 3-10.			"2) In this sense, I recommend to start with the broader discussion of the natural contract (beggining of session 3), ending with the question of how to put in practice this social contract, what challenges (what is the main focus of this text). The line 160 present this question as: ""The big question for steering is: how to initiate and accelerate system changes whose features are not perfectly clear and often times underdeveloped compared to the existing systems? This would make room to bring session 02 that addresses evolutionary/transformative governance to the TSEI.”"	3	2	su14052976_perova	0
+It is important that the authors give more emphasis on the scientific evidences of what they defend and anchor their argumentation in such evidences, whether theoretical or empirical.			"2) The section 02 """"Transformation pleas and approach to achieve this"""" can be improved."" I recommend reviewing the title referring to the governance debate that is addressed in the section, for example: ""Transformative pleas and approaches of governance to achieve this""...It is also important to better relate/ differentiate the evolutionary governance proposed by authors and the others discussed in the literature (adaptive, reflexive, deliberative, collaborative, etc.). The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution. I suggest that the authors present the debate and the different definitions and then present the notion of GE, relating it to the TSEI.”"	3	2	su14052976_perova	0
+How does a project mission contribute to a broader social contract?			3) The examples placed in section 03 could compose a section in itself, which sought to relate theory and practical examples (not cases), illustrating the proposed model. It will be nice to put forward the discussion of the conceptual variables of transformative governance proposed.	3	2	su14052976_perova	0
+One wonders whether the notion of transformative change really warrant this evolutionary perspective.			4) It remains not clear the relationship between the propositions presented by the authors in section 4 and the discutions in other sections. I suggest to improve the interface between the propositions presented and what comes before in the text, specialy linking with the contributions and limites of the proposal to face this challenges.	3	2	su14052976_perova	0
+To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy.			5) Formal aspects: The text needs a language revision and formatting of the figures to make them more readable. Avoid the excessive use of acronyms and review them throughout the text.	3	2	su14052976_perova	0
+We fully agree that there is not a single path or model for building public governance that favors sustainability.			Best regards I thank the authors and the editor for this opportunity to review and discuss this interesting paper and I hope that my comments help to improve the paper.	3	2	su14052976_perova	0
+The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.			The abstract is very confusingly written and needs a lot of editing. This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.	1	2	su14052981_makarova	0
+Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19.			I have some theoretical questions about the applicability of these approaches outside of Korea. For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.) I think many cities and countries will not be comfortable with this approach. I think this needs some exploration.	1	2	su14052981_makarova	0
+1 Dear Reviewer #2, I would like to thank Reviewer for the issues raised and for the recommendations.			I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned. I think that is both a big oversight, and an opportunity for critics. I think that piece needs to be included and worked into any assessment of smart cities and pandemics. Additionally, I've included a few other pieces on COVID and smart cities.	1	2	su14052981_makarova	0
+However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure.			Confusing or awkward wording in the abstract: “Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense? Just meaning sequential?)	1	2	su14052981_makarova	0
+“ 1) Confusing or awkward wording in the abstract: The text has been modified as follows: #1.			“a method responding in-time sequential by flexibly combining”	1	2	su14052981_makarova	0
+Table 7 does not bring much useful insights as well in its current form.			“Second, flexible smart city services are combined and deleted as needed to cope with COVID-19 has 17 been summarized. Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information. Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.” 2)	1	2	su14052981_makarova	0
+Are there concrete examples of such systems being built and operated, then just turned off?			(pg3) Tables 1 is interesting, but it’s not clear the source of the data. Is this from the literature? (if so, needs sourcing), or a synthesis by the authors? This may be because of the error message in sourcing in the document I received.	1	2	su14052981_makarova	0
+“a method responding in-time sequential by flexibly combining”			(pg4) Table 3 isn’t clear to me. What is “EA”? Not spelled out.	1	2	su14052981_makarova	0
+Sharifi, A., Khavarian-Garmsir, A. R., & Kummitha, R. K. R. (2021).			(pg5) Table 4 is very useful.	1	2	su14052981_makarova	0
+Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.”			(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen? Are there concrete examples of such systems being built and operated, then just turned off? Who would oppose turning them off? (I’d imagine many stakeholders) 3)	1	2	su14052981_makarova	0
+Are there concrete examples of such systems being built and operated, then just turned off?			Monitoring of sewage/wastewater for COVID case estimation Farkas, K., Hillary, L. S., Malham, S. K., McDonald, J. E., & Jones, D. L. (2020). Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19. Current Opinion in Environmental Science & Health, 17, 14-20. Daughton, C. G. (2020). Wastewater surveillance for population-wide Covid-19: the present and future. Science of the Total Environment, 736, 139631. Bogler, A., Packman, A., Furman, A., Gross, A., Kushmaro, A., Ronen, A., ... & Bar-Zeev, E. (2020). Rethinking wastewater risks and monitoring in light of the COVID-19 pandemic. Nature Sustainability, 3(12), 981-990. Jaiswal, R., Agarwal, A., & Negi, R. (2020). Smart solution for reducing the COVID‐19 risk using smart city technology. IET Smart Cities, 2(2), 82-88.	1	2	su14052981_makarova	0
+However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard.			I think the paper, and especially the abstract, are much improved. Both in terms of readability and accessibility, but also in terms of context and framing. I think the paper has the ability to make a contribution to the literature in a way that the previous version may not have been able to.	3	2	su14052981_makarova	0
+There are referencing issues with sections as well (e.g.			I think mentioning the scenario approach in the abstract might be worthwhile, but I don't think it's a huge problem if it isn't mentioned. Either way, I think the paper is in much better shape.	3	2	su14052981_makarova	0
+1 Dear Reviewer #2, I would like to thank Reviewer for the issues raised and for the recommendations.			This article advocates for better flexibility (in the sense of adding and removing when necessary) of smart city services and better integration between them. The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model. First, I have a series of concerns regarding the form. There are some phrasing issues and typos throughout the paper.	1	2	su14052981_makarova	0
+It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.			There are referencing issues with sections as well (e.g. line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic. For many references (e.g. [15], [22], [25]) it is impossible to retrieve the source. For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source. All these issues could have been picked up by a simple check. Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.	1	2	su14052981_makarova	0
+“a method responding in-time sequential by flexibly combining”			The content of the paper raises major concerns as well. In the introduction, I could not understand the motivation of the paper. The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic. However, I am lacking a concrete example of failure that proves the authors’ point. It is well-known that different services are hard to integrate together (this is not specific to the smart city), but I would have liked the authors to illustrate in light of the pandemic context, and maybe to the context of Korea since their contribution is specific to this geographic context in that it integrates the 12 smart city categories of Korea and the healthcare phases of Korea. The contribution is not presented in the introduction, only the problem is.	1	2	su14052981_makarova	0
+However, I am lacking a concrete example of failure that proves the authors’ point.			The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper. It is also hard to read, as it contains more tabular content than text. In Table 3, what is an EA?	1	2	su14052981_makarova	0
+It is well-known that many standards exist and are not used in every service, which causes integration issues.			Table 4 presents smart city projects of several different countries. It the threats concern COVID only, it does not really make sense, as many of the mentioned services simply do not have this goal (e.g. smart grid). If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature. The authors than write that hese services have a limitation in that cannot predict and prevent transmission of threats such as COVID.	1	2	su14052981_makarova	0
+From the additional explanations in the text, I understand that the idea is to track the movements of everyone and then use that information to trace back the contacts of confirmed infected patients.			Table 5 is useless in my opinion. It is well-known that many standards exist and are not used in every service, which causes integration issues. The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.	1	2	su14052981_makarova	0
+This article advocates for better flexibility (in the sense of adding and removing when necessary) of smart city services and better integration between them.			Section 2.3 has the same title as Section 2.2.	1	2	su14052981_makarova	0
+Additionally, I've included a few other pieces on COVID and smart cities.			It presents phased plans for pandemics. Several different plans are described, which is useless information for the reader. Indeed, the phases in a Texan county are of no use to understand the authors’ contribution. They could easily break the section down to the essential information that would be the WHO phases and the Korean healthcare phases, and mention that there are global and local plans throughout the world with different phases. Tables 8 and 9 can thus be removed. Table 7 is not very informative, all the phases say that actions from the previous phase should be continued or initiated. I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors. A case study approach restricted to Korea would be more informative and more consistent with the contribution of the paper, which is as I mentioned specific to Korea.	1	2	su14052981_makarova	0
+The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model.			Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find. I suggest e.g. “Revised model of smart city service structure”.	1	2	su14052981_makarova	0
+Smart solution for reducing the COVID‐19 risk using smart city technology.			The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV. However, it misses a critical point of COVID. Infected people are in the public space without knowing that they have COVID, since contagion happens before symptoms appears. Once infected people have been identified, they are supposedly in quarantine. Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts? Or do the authors want to check if infected people are outside their home instead of in quarantine? This is not clear to me. Also, and more importantly, the integration and flexibility aspects are not really addressed in the end. The authors do not explain how the services they propose could be integrated with existing ones (e.g. which standards should be used) and how the services can be added or removed. This part is only represented as clouds in the proposed structure model but not discussed further. Therefore, it seems that the contribution breaks down to proposing strict tracking services which feasibility and acceptance by the population is not assessed.	1	2	su14052981_makarova	0
+As advised by the reviewer, the content of the scenario was additionally reflected in the abstract.			My last concern is the ethical aspects of the proposed services. The authors acknowledge that not addressing these is a limitation of their research. In my opinion, this is more than a limitation, this is essential to discuss given the nature of what the authors propose.	1	2	su14052981_makarova	0
+Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19.			In summary, my suggestions are to focus the paper on the Korean context, in an in-depth case study approach where authors detail more their contribution, clearly illustrate what the problem is and how their contribution helps solving it.	1	2	su14052981_makarova	0
+The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement.			The authors made substantial efforts to take my comments into account, and I commend them for that. However, despite the good improvements, I have some remaining concerns.	3	2	su14052981_makarova	0
+Thank you once again for your pertinent suggestions and feedback.			"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version. Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography. Please feel free to add them back if you feel they should be included."	3	2	su14052981_makarova	0
+There are some phrasing issues and typos throughout the paper.			The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement. I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable. Indeed, if my understanding is correct, they are the factors that cause the limitations of smart city services the authors want to address.	3	2	su14052981_makarova	0
+[15], [22], [25]) it is impossible to retrieve the source.			The authors made some efforts to streamline Section 2, and it is much easier to follow in its current form. Table 3 just shows that medical, crime prevention, transportation, and environmental services are the most frequent, which is already said in the text, but without an explanation of what pilot city, regulatory sandbox, and smart city challenge are, it is difficult to extract anything else from this table. Table 7 does not bring much useful insights as well in its current form. The authors have not responded to my previous comment on this table, I thus redirect them to my review of the previous version. However, some parts still appear unnecessary to me.	3	2	su14052981_makarova	0
+Indeed, the phases in a Texan county are of no use to understand the authors’ contribution.			Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors. I would thus include it in Section 2 instead. Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2. It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.	3	2	su14052981_makarova	0
+(if so, needs sourcing), or a synthesis by the authors?			From the additional explanations in the text, I understand that the idea is to track the movements of everyone and then use that information to trace back the contacts of confirmed infected patients. I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations. The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.	3	2	su14052981_makarova	0
+This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.			Finally, and this is my biggest remaining concern, the authors have not responded to my comment regarding the lack of explanations on how the flexible adding and removal of the proposed services would be achieved. This is still unclear to me in the paper. This is a very important point, since it changes what is the contribution of the paper. The current contribution seems to be the proposal of new services to handle the COVID situation, or similar situations in the future. However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure. In other terms, what can we use from the authors' paper to achieve a more flexible smart city service structure?	3	2	su14052981_makarova	0
+I think the paper has the ability to make a contribution to the literature in a way that the previous version may not have been able to			In brief, the authors have made major improvements on the form and good improvements to the content. However, there are still several of my previous comments that were not answered or not answered in a sufficiently convincing way.	3	2	su14052981_makarova	0
+Table 3 just shows that medical, crime prevention, transportation, and environmental services are the most frequent, which is already said in the text, but without an explanation of what pilot city, regulatory sandbox, and smart city 2 challenge are, it is difficult to extract anything else from this table.			"The whole manuscript is full of ""Error! reference source..."" (e.g. line 85)"	1	2	su14052981_makarova	0
+Finally, and this is my biggest remaining concern, the authors have not responded to my comment regarding the lack of explanations on how the flexible adding and removal of the proposed services would be achieved.			Line 142: lack of flexibility	1	2	su14052981_makarova	0
+Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.			Please always explain abbreviations before use - e.g. line 149	1	2	su14052981_makarova	0
+The content of the paper raises major concerns as well.			Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020	1	2	su14052981_makarova	0
+I have some theoretical questions about the applicability of these approaches outside of Korea.			Lines 352-354: here we have two times Figure 2. Moreover, please bring the text together.	1	2	su14052981_makarova	0
+"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version."			In this source you can find a whole list of smart city technologies http://www.sinfonia-smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work	1	2	su14052981_makarova	0
+In brief, the authors' changes and explanations are a bit light for what I expected from a major revisions recommendations, but they added some nice clarifications that, for the most part, satisfy my concerns.			The abstract is very confusingly written and needs a lot of editing. This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.	1	2	su14052981_perova	0
+There are also many in-text reference errors with tables, which makes it hard to follow given the high number of tables (13!).			I have some theoretical questions about the applicability of these approaches outside of Korea. For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.) I think many cities and countries will not be comfortable with this approach. I think this needs some exploration.	1	2	su14052981_perova	0
+As a result, the critical point of being identified before an infectious disease spreads was missed.			I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned. I think that is both a big oversight, and an opportunity for critics. I think that piece needs to be included and worked into any assessment of smart cities and pandemics. Additionally, I've included a few other pieces on COVID and smart cities.	1	2	su14052981_perova	0
+Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers.			“Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense? Just meaning sequential?)	1	2	su14052981_perova	0
+Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately.			“a method responding in-time sequential by flexibly combining” “Second, flexible smart city services are combined and deleted as needed to cope with COVID-19 has 17 been summarized.	1	2	su14052981_perova	0
+For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.)			Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information. Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.” 2) Theoretical/Substantive questions: “Second, flexible smart city services are combined and deleted as needed to cope with COVID 19 has 17 been summarized.	1	2	su14052981_perova	0
+This is the place where the story must be told as simply and clearly as possible, and I think a lot of the abstract is confusing and hard to interpret.			(pg3) Tables 1 is interesting, but it’s not clear the source of the data. Is this from the literature? (if so, needs sourcing), or a synthesis by the authors? This may be because of the error message 3 in sourcing in the document I received.	1	2	su14052981_perova	0
+Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers.			(pg4) Table 3 isn’t clear to me. What is “EA”? Not spelled out.	1	2	su14052981_perova	0
+1 Dear Reviewer #1, I am grateful for your suggestions regarding my paper, and my answers to your comments are provided below.			(pg5) Table 4 is very useful.	1	2	su14052981_perova	0
+- “This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.” #4.			(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen? Are there concrete examples of such systems being built and operated, then just turned off? Who would oppose turning them off? (I’d imagine many stakeholders) 3) Key missing components/literatures:	1	2	su14052981_perova	0
+The authors made some efforts to streamline Section 2, and it is much easier to follow in its current form.			Smart City and COVID	1	2	su14052981_perova	0
+First, I have a series of concerns regarding the form.			Monitoring of sewage/wastewater for COVID case estimation	1	2	su14052981_perova	0
+The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV.			I think the paper, and especially the abstract, are much improved. Both in terms of readability and accessibility, but also in terms of context and framing. I think the paper has the ability to make a contribution to the literature in a way that the previous version may not have been able to.	3	2	su14052981_perova	0
+If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature.			I think mentioning the scenario approach in the abstract might be worthwhile, but I don't think it's a huge problem if it isn't mentioned. Either way, I think the paper is in much better shape.	3	2	su14052981_perova	0
+If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature.			This article advocates for better flexibility (in the sense of adding and removing when necessary) of smart city services and better integration between them. The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model. First, I have a series of concerns regarding the form. There are some phrasing issues and typos throughout the paper.	1	2	su14052981_perova	0
+Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find.			There are referencing issues with sections as well (e.g. line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic. For many references (e.g. [15], [22], [25]) it is impossible to retrieve the source. For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source. All these issues could have been picked up by a simple check. Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.	1	2	su14052981_perova	0
+Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately.			The content of the paper raises major concerns as well. In the introduction, I could not understand the motivation of the paper. The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic. However, I am lacking a concrete example of failure that proves the authors’ point. The contribution is not presented in the introduction, only the problem is. It is well-known that different services are hard to integrate together (this is not specific to the smart city), but I would have liked the authors to illustrate in 2 light of the pandemic context, and maybe to the context of Korea since their contribution is specific to this geographic context in that it integrates the 12 smart city categories of Korea and the healthcare phases of Korea.	1	2	su14052981_perova	0
+Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper. It is also hard to read, as it contains more tabular content than text. In Table 3, what is an EA?	1	2	su14052981_perova	0
+If the answer is yes, the official should seek approval from relevant authorities to get access to the data.			Table 4 presents smart city projects of several different countries. It the threats concern COVID only, it does not really make sense, as many of the mentioned services simply do not have this goal (e.g. smart grid). If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature. The authors than write that hese services have a limitation in that cannot predict and prevent transmission of threats such as COVID.	1	2	su14052981_perova	0
+Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography.			Table 5 is useless in my opinion. It is well-known that many standards exist and are not used in every service, which causes integration issues. The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.	1	2	su14052981_perova	0
+Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			Section 2.3 has the same title as Section 2.2.	1	2	su14052981_perova	0
+The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic.			It presents phased plans for pandemics. Several different plans are described, which is useless information for the reader. Indeed, the phases in a Texan county are of no use to understand the authors’ contribution. They could easily break the section down to the essential information that would be the WHO phases and the Korean healthcare phases, and mention that there are global and local plans throughout the world with different phases. Tables 8 and 9 can thus be removed. Table 7 is not very informative, all the phases say that actions from the previous phase should be continued or initiated. I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors. A case study approach restricted to Korea would be more informative and more consistent with the contribution of the paper, which is as I mentioned specific to Korea.	1	2	su14052981_perova	0
+However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard.			Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find. I suggest e.g. “Revised model of smart city service structure”.	1	2	su14052981_perova	0
+Thank you again for your supportive review to improve the manuscript.			The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV. However, it misses a critical point of COVID. Infected people are in the public space without knowing that they have COVID, since contagion happens before symptoms appears. Once infected people have been identified, they are supposedly in quarantine. Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts? Or do the authors want to check if infected people are outside their home instead of in quarantine? This is not clear to me. Also, and more importantly, the integration and flexibility aspects are not really addressed in the end. The authors do not explain how the services they propose could be integrated with existing ones (e.g. which standards should be used) and how the services can be added or removed. This part is only represented as clouds in the proposed structure model but not discussed further. Therefore, it seems that the contribution breaks down to proposing strict tracking services which feasibility and acceptance by the population is not assessed.	1	2	su14052981_perova	0
+line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic.			My last concern is the ethical aspects of the proposed services. The authors acknowledge that not addressing these is a limitation of their research. In my opinion, this is more than a limitation, this is essential to discuss given the nature of what the authors propose.	1	2	su14052981_perova	0
+I think mentioning the scenario approach in the abstract might be worthwhile, but I don't think it's a huge problem if it isn't mentioned.			In summary, my suggestions are to focus the paper on the Korean context, in an in-depth case study approach where authors detail more their contribution, clearly illustrate what the problem is and how their contribution helps solving it.	1	2	su14052981_perova	0
+In Smart Cities for Technological and Social Innovation (pp.			The authors made substantial efforts to take my comments into account, and I commend them for that. However, despite the good improvements, I have some remaining concerns.	3	2	su14052981_perova	0
+The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement.			"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version. Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography. Please feel free to add them back if you feel they should be included."	3	2	su14052981_perova	0
+Ministry of Land, Infrastructure and Transport (Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data.			The motivation of the paper is still unclear for me in the introduction, and the reasons for the limitations of smart city services are not clear in Section 2, but is much better explained later in the paper, which is a nice improvement. I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable. Indeed, if my understanding is correct, they are the factors that cause the limitations of smart city services the authors want to address.	3	2	su14052981_perova	0
+Sharifi, A., Khavarian-Garmsir, A. R., & Kummitha, R. K. R. (2021).			The authors made some efforts to streamline Section 2, and it is much easier to follow in its current form. Table 7 does not bring much useful insights as well in its current form. The authors have not responded to my previous comment on this table, I thus redirect them to my review of the previous version. However, some parts still appear unnecessary to me. Table 3 just shows that medical, crime prevention, transportation, and environmental services are the most frequent, which is already said in the text, but without an explanation of what pilot city, regulatory sandbox, and smart city 2 challenge are, it is difficult to extract anything else from this table.	3	2	su14052981_perova	0
+would it be constrained by laws and norms elsewhere?)			Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors. I would thus include it in Section 2 instead. Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2. It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.	3	2	su14052981_perova	0
+COVID-19 Scenario: Outbreak of COVID-19 in Korea ) scenario assumed by this study is uncertain.			From the additional explanations in the text, I understand that the idea is to track the movements of everyone and then use that information to trace back the contacts of confirmed infected patients. I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations. The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.	3	2	su14052981_perova	0
+I still encourage the authors to have a deep reflection on the legal and population acceptance of their proposal, but this can be the goal of another paper.			Finally, and this is my biggest remaining concern, the authors have not responded to my comment regarding the lack of explanations on how the flexible adding and removal of the proposed services would be achieved. This is still unclear to me in the paper. This is a very important point, since it changes what is the contribution of the paper. The current contribution seems to be the proposal of new services to handle the COVID situation, or similar situations in the future. However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure. In other terms, what can we use from the authors' paper to achieve a more flexible smart city service structure?	3	2	su14052981_perova	0
+However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure.			In brief, the authors have made major improvements on the form and good improvements to the content. However, there are still several of my previous comments that were not answered or not answered in a sufficiently convincing way.	3	2	su14052981_perova	0
+The authors than write that these services have a limitation in that cannot predict and prevent transmission of threats such as COVID.			"The whole manuscript is full of ""Error! reference source..."" (e.g. line 85)"	1	2	su14052981_perova	0
+1 Dear Reviewer #2, I would like to thank Reviewer for the issues raised and for the recommendations.			Line 142: lack of flexibility	1	2	su14052981_perova	0
+That said, I think it requires some tweaks to make it as effective as it could be.			Please always explain abbreviations before use - e.g. line 149 Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020	1	2	su14052981_perova	0
+Regarding the form, the reference errors have been removed, but the line breaks when referring to figures are still there.			Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020.	1	2	su14052981_perova	0
+Bogler, A., Packman, A., Furman, A., Gross, A., Kushmaro, A., Ronen, A., ... & Bar-Zeev, E. (2020).			Lines 352-354: here we have two times Figure 2. Moreover, please bring the text together.	1	2	su14052981_perova	0
+It would allow a smoother transition between Section 2 and Section 3, as the modified Section 2 would conclude with the need for a novel smart city service structure.			In this source you can find a whole list of smart city technologies http://www.sinfonia-smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work	1	2	su14052981_perova	0
+Point 2: Are the research design, questions, hypotheses and methods clearly stated?			Thank you for giving the opportunity to read such an interesting paper.	1	2	su14095590_perova	0
+1 Response to Reviewer 5 Comments Point 1: Title: The title demands a review because there isn’t a connection between the first part and the second one.			For a better and accurate model I recommend the authors to eliminate from the model the extreme value. For example in Fig 2. - workers values greater then 40, community greater than 58, environment 45, customers 46 (the bullets), in Fig 3 - values with global score greater than 130, in Fig. 5 also all the bullets. Then repeat all the statistical analysis.	1	2	su14095590_perova	0
+Literature review: There is still work to do discussing the five areas, not just relating them to each dimension of the Triple Bottom Line of the sustainability concept.			In lines 391-395 I would prefer an Sk and Ku between -1 and 1. You have some values that are not included in this interval - maybe because of the outliers from the figures above that I recommend to exclude.	1	2	su14095590_perova	0
+We have made all the minor corrections indicated.			The paper is interesting; however, it is not clear what the authors are trying to achieve with this study. The authors test a popular assessment tool using confirmatory factor analysis and finds that the model is weak. However, there is no in-depth discussion or recommendations that follow. In fact, the discussion needs to be grounded in previous research findings in regard to CSR/sustainability indices/assessment tools/initiatives. Unfortunately, I am really not sure what the point is here.	1	2	su14095590_perova	0
+It definitely adds a value to the existed literature.			Further, the hypothesis does not seem to be written correctly. First, it should be stated prior to the methods section. Second, currently it is descriptive. In other words, it doesn't state what the authors are trying to test in this study.	1	2	su14095590_perova	0
+1 Response to Reviewer 3 Comments (round 2) Point 1:			Unfortunately, I cannot recommend publication unless major changes are made to the discussion section and possibly recommendations are added for future research and practice.	1	2	su14095590_perova	0
+It’s clever to reduce the number of companies, though it isn’t clear who contacts the companies, the authors or Company B?			English language and style are fine/minor spell check required .	3	2	su14095590_perova	0
+The authors should reflect the presence of that in the text.			Are the arguments and discussion of findings coherent, balanced and compelling?	3	2	su14095590_perova	0
+Some authors were added, but for me, this section demands an improvement to enhance the quality of the text, and to turn it into a scientific paper.			Are the conclusions thoroughly supported by the results presented in the article or referenced in secondary literature?	3	2	su14095590_perova	0
+Point 3: Introduction: This section demands more attention because there are some empty spaces to fill.			The paper is well structured and presented in a very meaningful manner. It definitely adds a value to the existed literature.	1	2	su14095590_perova	0
+There isn’t a source below the Title of Figure 1, though after that seems that B Corporation is the owner of it.			Line 169 page 4, studies [28] & [29] do not reference to any presented argument(s) unless the preceding para.	1	2	su14095590_perova	0
+If the rules of the Journal don’t allow to write down the Figures who is the author, I suggest to authors write it in a paragraph before the Fig 1, or in a footnote, according to Editor’s allowance.			The word of Table in all text should start with capital letter (e.g. Line 199, L. 223)	1	2	su14095590_perova	0
+Suddenly, appears one in the Method section, in a paper with a limited presentation of a technical model/certificate from an existed company.			Paragraph presented after Table 2 (page 7) is a repetitive content to what has been presented in the Table. It could be removed or move to footnote.	1	2	su14095590_perova	0
+I accessed the link available in line 191 and there was a message ‘404 page not found, but when I clicked on ‘What is B Corporation’, I found the ‘areas’ and the steps for a company that intends to assess its practices and be certified.			It is better to show the P-values in Table 3 rather than (***) although a note is indicated in this regard.	1	2	su14095590_perova	0
+Method: For me, as commented in the first round, Table 2 is a repetition, doesn’t need to be in the text.			Very good practical recommendations and criticism to the B Corp certification have been presented in the last para. of section (6). No any future research recommendations have been suggested by the respected authors.	1	2	su14095590_perova	0
+Response 5: We believe that the reviewers' suggestions have been very helpful in improving the manuscript.			Is the content succinctly described and contextualized with respect to previous and present theoretical background and empirical research (if applicable) on the topic? Must be improved Response 4: Thanks for the comment.	3	2	su14095590_perova	0
+Seems that verb ‘improve’ isn’t appropriate for ‘impact’, which leads the reader to understand that these impacts are negative, which isn’t true.			Can be improved. Are the research design, questions, hypotheses and methods clearly stated?	3	2	su14095590_perova	0
+The analysis is interesting, but demands more comparisons and explanations for cuts and adds.			The text has some improvements, but there are some points that still demand more attention.	3	2	su14095590_perova	0
+In the first round, I observed and commented that the link in line 191 wasn’t working.			The title demands a review because there isn’t a connection between the first part and the second one. We understand what the authors intend to inform us after reading the paper. Even though this was clear to me, the title doesn’t show the real idea developed in the paper. The objective is a good guide for the title.	1	2	su14095590_perova	0
+Point 3: Introduction: This section demands more attention because there are some empty spaces to fill.			The content is well organized, but the results need some to attract the reader. What is the main result of the B assessment with the confirmatory analysis? Besides, the contribution isn’t clear here.	1	2	su14095590_perova	0
+Point 3: Unfortunately, I cannot recommend publication unless major changes are made to the discussion section and possibly recommendations are added for future research and practice.			This section demands more attention because there are some empty spaces to fill. The authors should explain better what is B Impact Assessment. What is its contribution to the structure of a model using Confirmatory Analysis? For me, lines 39 and 40 show the aim of the paper, but the idea is different from the ones presented in the abstract (“The main objective is to understand the B Impact Assessment, verifying its added value in the sustainability process and socio-economic development of the business sector”). The sentence in the Introduction is more complete than the main objective in the Abstract. The ‘areas’ represent a broader goal than the main dimensions of sustainability (Triple Bottom Line).	1	2	su14095590_perova	0
+Thus, to ensure the 3 consistency of the database and the statistical analysis subsequently performed, we reduced the database to a recent period in which the same measurement model was used.” Results: I didn’t see ads in Cronbach’s alpha analysis (the text is the same as the first version, see lines 457-486 – new version and compare with an old version from line 405- 433), though there are some adjustments in their numeric results.			The lines from 70 to 82 could be summarized, and cut and paste in the Introduction section. I suggest cutting the text from lines 90 to 103 because it’s quite an advertisement for the B Certificate. The content in this section is exclusively dedicated to the Certification and the Corporation which offers it, there isn’t any scientific information there. The section demands a literature review about the theme, as the relationship between the certificate with adding value, sustainability, and the ‘areas’ governance, workers, community, environment, and customers. What is the idea of the assessment of sustainability, in these areas? This discussion is the basis of the paper, related to the objective, and connected with Figure 1 presented in the next section, Method. There isn’t a literature review in this work. Even 2 though the areas are from B Lab, the authors should study and related them to sustainability and value creation.	1	2	su14095590_perova	0
+What is the main result of the B assessment with the confirmatory analysis?			In the Abstract, the objective added an economic dimension to the development, but in lines 180 and 181 the impact to be measured includes just environmental and social. It was not clear to me who is the author of Figure 1, the B Lab, or the authors of this paper? There isn’t a source below the Title of Figure 1, though after that seems that B Corporation is the owner of it. The source of data from all companies in the sample isn’t clear, is B Corporation (site) or authors contact all these companies and did the survey? I understood that the data is from the B Corporation website. I accessed the link available in line 191 and there was a message ‘404 page not found, but when I clicked on ‘What is B Corporation’, I found the ‘areas’ and the steps for a company that intends to assess its practices and be certified. The authors should develop more theory than describe a model which has already existed. Table 2 and the paragraph after it is a repetition of Figure 1 and Table 1, could be cut. It’s clever to reduce the number of companies, though it isn’t clear who contacts the companies, the authors or Company B? The hypothesis is interesting when they come from a discussion in the literature review to the field research to test a theory (models, methods, frameworks, etc). Suddenly, appears one in the Method section, in a paper with a limited presentation of a technical model/certificate from an existed company.	1	2	su14095590_perova	0
+Point 6: Results: The sentence from line 137 to line 138 isn’t clear when I saw Figure 6.			The sentence from line 137 to line 138 isn’t clear when I saw Figure 6. The authors could use a standard for a plural of index word, ‘indices’ (345) or ‘indexes’ (362). In line 384 the authors should substitute ‘chapter’ for ‘section’. The authors presented and analyzed the variables/areas of certification with many models, it is interesting and is a basis for analysis in the next section.	1	2	su14095590_perova	0
+There are minor corrections to be considered.			In the first sentence, what is the topic that the authors mentioned? What source provided that information? The analysis is interesting, but demands more comparisons and explanations for cuts and adds. For me, the answer to Hypothesis 1 isn’t clear. The authors should reflect the presence of that in the text. The Cronbach’s Alpha ‘selected’ just ‘environment’ area of the B Lab model. This is an interesting result, which could be more analyzed with other assessment models presented in the text, to say what the authors wrote at the end of the section.	1	2	su14095590_perova	0
+The sentence in the Introduction is more complete than the main objective in the Abstract.			Conclusion: I just see one objective, but in the first sentence the authors wrote ‘objectives’. Where are they? The suggestions are interesting, but demand more analysis in the previous section.	1	2	su14095590_perova	0
+Below we provide the details on how your comments and suggestions have been addressed.			1- Need to update the literature review with recent studies	1	2	su14127346_makarova	0
+We take this review as a valuable opportunity to foster the quality of the paper.			2- Please add a theoretical background section	1	2	su14127346_makarova	0
+But you are working one sector only, aren't you?			3- What are the policy implications of your study	1	2	su14127346_makarova	0
+Similar problems might exist for a negative tobin's Q.			4- Add discussion of results section and relate your results with theories.	1	2	su14127346_makarova	0
+Similar problems might exist for a negative tobin's Q.			"""To the best of our knowledge, this is the only study that addresses the role of economic and political uncertainty as a moderator in a model that accounts for the dynamic patterns of firm environmental performance"" (line 128). --> I believe the authors intended ""moderating role."""	1	2	su14127346_makarova	0
+The authors could go into greater detail to explain the logic behind this relationship.			The authors could go into greater detail to explain the logic behind this relationship. As it stands, the paper offers little insight into the reasoning behind their hypothesis.” We answered this comment along different dimensions.	1	2	su14127346_makarova	0
+International Journal of Financial Studies, 10(1), 15.			It would be useful if the authors provided some speculative thoughts on the finding that the slope of the GenDiv variable is negative in highly uncertain countries on the firm's environmental performance (Figure 3 (c) red line).	1	2	su14127346_makarova	0
+Overall, I believe that this study has a high potential to add to the literature on the relationship between gender diversity and firm environmental performance.			1- In lines 28 and onwards, you argue a lack of literature looking at dynamic (GMM) analysis of the role of female on CSR and ESG issues, but fail to mention some examples. Please provide some, and position your work in that (brief) strand of literature. One example you should consider is Gaio, C., & Gonçalves, T. C. (2022). Gender diversity on the board and firms’ corporate social responsibility. International Journal of Financial Studies, 10(1), 15.	1	2	su14127346_makarova	0
+The arguments above provides a rationale for the worsened environmental performances of firms in response to higher uncertainties, and explains the gap in effects for firms across countries of different uncertainty levels.			2- Please revise if it makes sense to present your research hypothesis in the introduction.	1	2	su14127346_makarova	0
+This suggests that an effective way to improve environmental quality and mitigate the adverse effects of economic activities is to encourage gender diversity in corporate boards in countries with low uncertainty where we find a higher impact on the firms’ environmental performances.			"3- Please reconsider the use of adjectives such as ""complicated"" (line 79), interestingly (line 88) and so on."	1	2	su14127346_makarova	0
+We investigate this finding further by examining, for different gender diversity levels, (a) the intensity of the relationship across high and low uncertainty levels and (b) the marginal impact of a country’s uncertainty status on the firm’s environmental performance (Figure 3).			"4- What do you mean by ""in the crowd"" (line 141). Please revise carefully english for clarity."	1	2	su14127346_makarova	0
+Regards, Pavagal Latha Fundamentals Content Support			5- Please revise your data: your max tangibility is over 2 (which means that Net PP&E, a subset of Assets, is 2 times those same Assets - this should be a typo. Similar problems might exist for a negative tobin's Q.	1	2	su14127346_makarova	0
+But you are working one sector only, aren't you?			"6 - In line 192 you mention ""all the sectors"". But you are working one sector only, aren't you?"	1	2	su14127346_makarova	0
+The gap in environmental performance between countries with high versus low uncertainty increases as gender diversity in corporate boards increases.			7- Explain your sentences in lines 242-244. What did you do specifically?	1	2	su14127346_makarova	0
+We revise and elaborate on the explanation and interpretation of the results.			"8- Lines 330 and onwards are presenting conclusions that are not warranted by the results and the english is a bit strange (what do you measnby ""households' anxiety""?). Similar problems stem from the following paragraphs discussing the results."	1	2	su14127346_makarova	0
+Among the three methods used, the upper bilateral floret injection (UBFI) maximizes DON accumulation in grain and significantly distinguishes DON content in different wheat genotypes, indicating that the method could be useful for breeding program to develop wheat cultivars with resistance to DON accumulation.			I would like to suggest authors label inoculation points in Figure 1, which could give readers a clear view of three different inoculation methods.	1	2	toxins14060409_makarova	0
+The paper is of high interest for the community working on FHB and shows the importance of inoculation method on the results of disease and mycotoxin accumulation.			Line 142: “proportion of PPSD” should be “proportion of PSD or PPSD”.	1	2	toxins14060409_makarova	0
+The following are some comments and suggestions.			Line 143: “PPSD-premature spike death” should be “PPSD-proportion of premature spike death”.	1	2	toxins14060409_makarova	0
+Mycotoxin deoxynivalenol (DON) contamination of wheat and other grains has been a concern of food safety.			Lines 168 - 169: Should it be that DON content of most lines without Fhb1 exceeded 1000 µg/kg under UBFI? Where is 4224.38 µg/kg in Table 1? Do you mean in Table S1?	1	2	toxins14060409_makarova	0
+I would like to recommend its acceptance for publication.			Lines 411 & 413: “PPSB” should be “PPSD”.	1	2	toxins14060409_makarova	0
+Point 3: Identity of the strain used for infection should be confirmed: a multilocus species characterisation is needed to confirm the species of the strain or a reference to a publication where the strain was described and appropriately characterised.			Table S1: “PPSD - premature spike death” in the footnote should be “PPSD - proportion of premature spike death”.	1	2	toxins14060409_makarova	0
+Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			Fhb1- and Fhb1+ don’t appear in Table S1, so they can be removed from the footnote.	1	2	toxins14060409_makarova	0
+Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			I would suggest to avoid citing supplementary figures at the beginning of the chapter. See for example chapter 2.4. Given that 2.4 chapter discusses data of supplementary figure 4 I would suggest to make figure S4 as Figure 4 of the manuscript.	1	2	toxins14060409_makarova	0
+The paper is of high interest for the community working on FHB and shows the importance of inoculation method on the results of disease and mycotoxin accumulation.			To futher clarify the infection methods a supplementary figure detailing graphically the modes of inoculation with figures would make the paper extremely useful for the community	1	2	toxins14060409_makarova	0
+Given that 2.4 chapter discusses data of supplementary figure 4 would suggest to make figure S4 as Figure 4 of the manuscript.			Identity of the strain used for infection should be confirmed: a multilocus species characterisation is needed to confirm the species of the strain or a reference to a publication where the strain was described and appropriately characterised.	1	2	toxins14060409_makarova	0
+1 Response to Reviewer 2 Comments Point 1: I would suggest to avoid citing supplementary figures at the beginning of the chapter.			Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication)	1	2	toxins14060409_makarova	0
+While there are important new insights provided by this work, the manuscript can be improved in terms of data presentation and referencing the existing literature on the IFITM proteins.			The largest concern is regarding the colocalization of IFITM3 and Rab11A. It seems (as the authors state) that these represent abortive events (Figure 7). Therefore the inclusion of the movie examining the colocalization of Rab11A GFP, IFITM3-SNAP, and labeled virion seems completely unnecessary. In addition, the movie does not provide any details on cellular features and a viewer is left wondering where the foci are coming from and going. This reviewer suggests removing the movie or alternatively including labels on the movie of cellular features and including a scale bar.	1	2	v11060548_makarova	0
+These IFITM3 clusters were found to correlate with restriction of virus infection and to co-localize with incoming virus.			b. The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?	1	2	v11060548_makarova	0
+Figure 2A: The siRNA control should be shown.			c. Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.	1	2	v11060548_makarova	0
+Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.			d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.	1	2	v11060548_makarova	0
+Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.	1	2	v11060548_makarova	0
+Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu.			Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.	1	2	v11060548_makarova	0
+Figure 3A: Why is there so much IFITM3 present in the uninfected cells?			Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.	1	2	v11060548_makarova	0
+In this study the authors examine the localization of IFITM3 during a viral infection in airway epithelial cells.			Figure 4: Uninfected control should be shown in the main text.	1	2	v11060548_makarova	0
+Figure 4: Label the figure so that it can be interpreted independent of the legend.			Figure 4: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_makarova	0
+Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity.			Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_makarova	0
+Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017).			Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.	1	2	v11060548_makarova	0
+The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.	1	2	v11060548_makarova	0
+The supplemental figures only include experiments that reproduce previously published work or are not absolutely necessary to support the major and most novel findings of the paper.			Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.	1	2	v11060548_makarova	0
+Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.			Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.	1	2	v11060548_makarova	0
+The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.	1	2	v11060548_makarova	0
+Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity.			Supplemental figures should be included in the main text.	1	2	v11060548_perova	0
+Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017).			The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining. At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?	1	2	v11060548_perova	0
+They convincingly shown that IFITM3 clusters in cells where NP is primarily in the cytoplasm in an interferon dependent manner.			Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.	1	2	v11060548_perova	0
+Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.			Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.	1	2	v11060548_perova	0
+The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.	1	2	v11060548_perova	0
+They convincingly shown that IFITM3 clusters in cells where NP is primarily in the cytoplasm in an interferon dependent manner.			Figure 2A: The siRNA control should be shown. qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.	1	2	v11060548_perova	0
+While there are important new insights provided by this work, the manuscript can be improved in terms of data presentation and referencing the existing literature on the IFITM proteins.			Figure 2B: These data need to be improved. The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.	1	2	v11060548_perova	0
+This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species.			Figure 3A: Why is there so much IFITM3 present in the uninfected cells? This does not appear to increase post infection as was shown previously.	1	2	v11060548_perova	0
+The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining.			Figure 4: Uninfected control should be shown in the main text.	1	2	v11060548_perova	0
+Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			Figure 4: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_perova	0
+At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.	1	2	v11060548_perova	0
+The authors have largely addressed my previous comments.			Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.	1	2	v11060548_perova	0
+There are few clarifications that would strengthen the overall manuscript.			Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity. Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.	1	2	v11060548_perova	0
+Overall it is an interesting study, using sophisticated microscopy techniques to examine the intracellular distribution of IFITM3 during viral infection.			Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2014; Li, JBC, 2015) Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza.	1	2	v11060548_perova	0
+At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			Line 65 states that subsequent studies have failed to confirm an association between SNP rs12252 and severe influenza. This may be true, but far more studies have confirmed this link than have refuted it. Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent. Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu. For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.	1	2	v11060548_perova	0
+Figure 3A: Why is there so much IFITM3 present in the uninfected cells?			Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017). This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species. Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.	1	2	v11060548_perova	0
+They also discussed their results in detail and put the difference in their results compared to the data with the literature.			The innovation of this research needs to be better explored.	1	2	w14030367_perova	0
+Response 15: Thanks very much for this comment.			Cite the following references: Panagopoulos, A. (2021). Energetic, economic and environmental assessment of zero liquid discharge (ZLD) brackish water and seawater desalination systems. Energy Conversion and Management, 235. Techno-economic assessment of Minimal Liquid Discharge (MLD) treatment systems for saline wastewater (brine) management and treatment. Process Safety and Environmental Protection, 146, pp. 656-669. Study and evaluation of the characteristics of saline wastewater (brine) produced by desalination and industrial plants. Environmental Science and Pollution Research, 1-14. Lines 33-38: You should mention that discharge of PPCPs degrades water quality and thus it cannot be directly used for potable water and industrial applications.	1	2	w14030367_perova	0
+I can see very well-defined experiments and produced good results.			Conclusion: Discuss the applicability of your findings/results and future study in this field.	1	2	w14030367_perova	0
+Response 13: Thank you very much for your kind reminder.			Conclusion: Make it as one or two paragraphs.	1	2	w14030367_perova	0
+Dear Authors, i reviewed again the revised version.			Table 1: What is the salinity (in mg/L) of the samples ?	1	2	w14030367_perova	0
+However, the long-term pilot-scale study is urge to be investigated.			However, there are some mistakes in the writing of the paper.	1	2	w14030367_perova	0
+The PPCPs catalog includes a variety of chemicals, such as human and veteri- 36 nary drugs, fragrances and disinfectants used in personal care products (e.g., soaps, lo- 37 tions, sun-screens and body cleaning products) and household chemicals [3].			The title seems good, but the abstract seems to be fine. Please add one problem statement line in abstract to justify this sentence ``the long-term pilot-scale study is urged to be investigated.``.	1	2	w14030367_perova	0
+: +86 15765532017; E-mail: chnwangjinlong@163.com (J. Wang) 14 Abstract： Biological activated carbon (BAC) biofilter coupling ultrafiltration (UF) is a promising 15 process for the treatment of surface water contaminated by pharmaceutical and personal care prod- 16 ucts (PPCPs).			Research gap should be delivered on more clear way with directed necessity for the future research work.	1	2	w14030367_perova	0
+Response 5: Thanks very much for this comment.			Introduction section must be written on more quality way, i.e., more up-to-date references addressed. Please target the specific gap such as 2015-2021 etc.	1	2	w14030367_perova	0
+Dear Authors, i reviewed again the revised version.			Page 1 Line 38. Please cite this reference with existing reference 3….Role of nanotechnology for design and development of cosmeceutical: application in makeup and skin care.	1	2	w14030367_perova	0
+Why is the DO concentration of BAC-effluent sometimes higher than the Raw water.			The novelty of the work must be clearly addressed and discussed, compare previous research with existing research findings and highlight novelty.	1	2	w14030367_perova	0
+The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent).			What is the main challenge? Why author choose this material? Please highlight in the introduction part.	1	2	w14030367_perova	0
+The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances.			Page 2 Line 55 need a reference. Please consider these at end of this sentence……The oxidation method exhibited a fast reaction speed and high removal efficiency…(i) Role of nanomaterials in the treatment of wastewater: A review (ii) Advances and challenges in developing efficient graphene oxide-based ZnO photocatalysts for dye photo-oxidation.	1	2	w14030367_perova	0
+Response 2: We agree with the suggestion of this reviewer.			The main objective of the work must be written on the more clear and more concise way at the end of introduction section.	1	2	w14030367_perova	0
+Two native English-speaking colleagues help us verify the manuscript.			Please check the abbreviations of words throughout the article. All should be consistent.	1	2	w14030367_perova	0
+##Comments on the title, Abstract, and References			Please add chemical reagents section and stated all chemical with brand specifications.	1	2	w14030367_perova	0
+The paper is interesting.I recommend publication only if the following issues can be addressed.			Regarding the replications, authors confirmed that replications of experiment were carried out. However, these results are not shown in the manuscript, how many replicated were carried out by experiment? Results seem to be related to a unique experiment. Please, clarify whether the results of this document are from a single experiment or from an average resulting from replications. If replicated were carried out, the use of average data is required as well as the standard deviation in the results and figures shown throughout the manuscript. In case of showing only one replicate explain why only one is shown and include the standard deviations.	1	2	w14030367_perova	0
+Point 7: Please show the t-value and p-value in a scientific view (e.g., 2.79E-4 would be P <.001).			Please revise your paper accordingly since some issue occurs on several spots in the paper. Please use Fig. or figure? It very confusing. Article should be in one pattern.	1	2	w14030367_perova	0
+The article topic is intriguing and promising in the area.			Please provide high quality image of figure	1	2	w14030367_perova	0
+Response 2: Thank you very much for your kind reminder.			Please use one style for units such as m3/h or m3h-1 Please revise your paper accordingly since some issue occurs on several spots in the paper.	1	2	w14030367_perova	0
+Overall, the manuscript can be accepted after the minor correction/revision to be published in the journal of Water.			Please add a comparative profile section to compare your results and prove how it better than previous.	1	2	w14030367_perova	0
+Is it true that the molecular weight of viruses and bacteria is more than that of PPCPs?			Section 4 should be renamed by Conclusion and Future perspectives. Conclusion section is missing some perspective related to the future research work, quantify main research findings, highlight relevance of the work with respect to the field aspect. In the present form conclusion is very weird.	1	2	w14030367_perova	0
+Raw water quality parameters of river water (significant value, 0.05).			To avoid grammar and linguistic mistakes, Major level English language should be thoroughly checked. Please revise your paper accordingly since several language issue occurs on several spots in the paper.	1	2	w14030367_perova	0
+Two native English-speaking colleagues help us verify the manuscript.			Please follow the journal guidelines. Reference formatting need carefully revision. All must be consistent in one formate.	1	2	w14030367_perova	0
+Please, clarify whether the results of this document are from a single experiment or from an average resulting from replications.			Authors should avoid abbreviations in the title and the abstract.	1	2	w14030367_perova	0
+Why is the DO concentration of BAC-effluent sometimes higher than the Raw water.			Authors may revise the title to include river water instead of surface water.	1	2	w14030367_perova	0
+Why is the standard deviation of Erythromycin showing a high value?			References should be according to the journal format.	1	2	w14030367_perova	0
+The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent).			Line 44-45: Ultrafiltration (UF) as emerging technology, has been widely used to remove pollutants such as particles, colloids, bacteria and viruses, reducing the risk of water-borne diseases and …UF membranes cannot effectively rejected these soluble substances”. Please clarify, why ultrafiltration cannot remove the PPCPs since it can remove bacteria and viruses.	1	2	w14030367_perova	0
+Point 3: Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10].			Please add t-test results in Table 1 for each parameter to understand the significant differences. Authors may provide data in the supporting information file.	1	2	w14030367_perova	0
+Response 9: Thank you very much for your kind reminder.			The reviewer suggests evaluating the statistical t-test for Figures 4.	1	2	w14030367_perova	0
+Response 9: Thanks very much for this comment.			t-test results should be included in Table Why is the standard deviation of Erythromycin showing a high value?	1	2	w14030367_perova	0
+Point 15: Please add a comparative profile section to compare your results and prove how it better than previous.			Please look at the curve of BAC/UF; there was a symbol missing in near 85 days. Please show each symbol.	1	2	w14030367_perova	0
+Point 16: Section 4 should be renamed by Conclusion and Future perspectives.			The conclusion may be revised.	1	2	w14030367_perova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			Authors should avoid abbreviations in the title. Authors should add a list of abbreviations before the references.	3	2	w14030367_perova	0
+The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances.			Please avoid repeating the full name and abbreviation throughout the manuscript if you used the first-time abbreviation.	3	2	w14030367_perova	0
+Size exclusion is considered the primary removal mechanism for the UF.			Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. Size exclusion is considered the primary removal mechanism for the UF. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite”. Sorry, but I don't see why UF can reject viruses and bacteria but not PPCPs. Is it true that the molecular weight of viruses and bacteria is more than that of PPCPs? Please elaborate.	3	2	w14030367_perova	0
+Response 4: Thank you very much for your kind reminder.			The authors should include some information regarding the biological degradation of PPCPs by microorganisms in the introduction section.	3	2	w14030367_perova	0
+Besides, the t-test was used and proved the significant difference between BAC-Effluent and BAC/UF-effluent.			Line 53-55: Have the authors compared the BAC-UF performance to that of nanofiltration?	3	2	w14030367_perova	0
+The paper is interesting.I recommend publication only if the following issues can be addressed.			I strongly suggest that the authors explain how they performed the t-tests. Please include the sample size (n) or degree of freedom (df).	3	2	w14030367_perova	0
+Point 2: Please avoid repeating the full name and abbreviation throughout the manuscript if you used the first-time abbreviation.			Please show the t-value and p-value in a scientific view (e.g., 2.79E-4 would be P <.001).	3	2	w14030367_perova	0
+Point 3: How many replications you performed for your experiments?			Line 161-176: Please delete the name of medicinal compounds not used for this study.	3	2	w14030367_perova	0
+- Lines 33-38: You should mention that discharge of PPCPs degrades water quality and thus it cannot be directly used for potable water (via desalination) and industrial applications.			1) was observed in Fig. 1(c). Why is the DO concentration of BAC-effluent sometimes higher than the Raw water. Please explain it in the manuscript. The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent). Point 6: Why the same trend of increase or decrease in graphs (Fig. 1) was observed in Fig. 1(c).	3	2	w14030367_perova	0
+Response 6: Thanks very much for this comment.			The same trend (Figure 2) was mainly due to the stable removal ability of BAC and UF for organics, causing the removal restriction. The periodic backwash (7days) of BAC caused sometimes the dissolved oxygen concentration of BAC-effluent higher than that of raw water. After the gas scrubbing and the hydraulic backwashing, the dissolved oxygen detection of the effluent was carried out, resulting in the above results for dissolved oxygen. Besides, the t-test was used and proved the significant difference between BAC-Effluent and BAC/UF-effluent. Response 6: Thanks very much for this comment.	3	2	w14030367_perova	0
+Please feel free to contact me if any questions.			There is no sampling procedure, depth, or random selection of the soils evaluated.	1	2	w14081258_makarova	0
+Worse more, the soil can not be brought to China.			Is the explanation of χ is for molecular connectivity indices or branching index?	1	2	w14081258_makarova	0
+Author Response Dear Editor： Thank you very much for the comments of my manuscript.			If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.	1	2	w14081258_makarova	0
+Haidian District, Beijing, 100048 P.R.China Tel.			The basic physi-chemical parameters of soils should be given.	1	2	w14081258_makarova	0
+Your careful review of this manuscript is highly appreciated.			The irrigation water physi-chemical parameters should be given.	1	2	w14081258_makarova	0
+Yours sincerely, Shiyu Wang [email protected] Author Response File: Author Response.docx			The kinect models should be given.	1	2	w14081258_makarova	0
+Yours sincerely, Shiyu Wang [email protected]			Figure 1 should be revised according to kinect model.	1	2	w14081258_makarova	0
+China Institute of Water Resources and Hydropower Research Chegongzhuang West Road NO.			I strongly recommend the authors to analysis the biomass of the soil samples.	1	2	w14081258_makarova	0
+In the future study, this would be taken into accounted.			"I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: ""The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars"" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?"	3	2	w14081258_makarova	0
+Comments I do not understand why soil samples were 0.3 mm sieved.			Figure 1 legends of axis need font size larger, please make readable the figuresR：	3	2	w14081258_makarova	0
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar.			There is no sampling procedure, depth, or random selection of the soils evaluated.	1	2	w14081258_perova	0
+Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu.			Is the explanation of χ is for molecular connectivity indices or branching index?	1	2	w14081258_perova	0
+In the future study, this would be taken into accounted.			The values obtained of I DW and steric hindrance are not explained.	1	2	w14081258_perova	0
+Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP.			The more critical issue lies in the analytical methodology used for isomers quantification. First of all, figure one is a chromatogram, not a spectrum. The chromatogram is not well resolved, i.e., there is no separation among the different analytes determined. For example, look at peaks 4, 5, and 6. There is no resolution among the isomers, the same for NP 7 and 8 and NP 8 and 10; thus, it is impossible to quantify them. If correct quantification is not assessed, then figure 1 and table 1 and 2 are not valid.	1	2	w14081258_perova	0
+Author Response Dear Editor： Thank you very much for the comments of my manuscript.			The basic physi-chemical parameters of soils should be given.	1	2	w14081258_perova	0
+Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu.			The irrigation water physi-chemical parameters should be given.	1	2	w14081258_perova	0
+"Moreover, in the subsequent sentence is reported: ""The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars""Did you sieve the soil twice at 0.3 mm and then 0.25 mm?"			The kinect models should be given.	1	2	w14081258_perova	0
+The following are the details of the revision and marked red in the “resubmit” version.			Figure 1 should be revised according to kinect model.	1	2	w14081258_perova	0
+Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104).			I strongly recommend the authors to analysis the biomass of the soil samples.	1	2	w14081258_perova	0
+气相色谱-质谱法检测地下水中12种对壬基酚同分异构体.分析化学，2013，41（11）：1699-1703.			"I do not understand why soil samples were 0.3 mm sieved. Why? The active soil components (lime, sand, clay) where microorganisms live and organic carbon is accumulated is just less than 2mm. Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.R： Moreover, in the subsequent sentence is reported: ""The soil was 0.25 mm-sieved to remove large particles; and then weighed series of 10 g aliquots into 250 mL brown jars"" Did you sieve the soil twice at 0.3 mm and then 0.25 mm?"	1	2	w14081258_perova	0
+Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu.			Figure 1 legends of axis need font size larger, please make readable the figuresR：	1	2	w14081258_perova	0
+I have tried to address your principle concern by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	This may be a difference in semantics, but I do not consider R2 the implementation of an extant case the same as an academic contribution in developing a qualitative case study.			admsci5030125_boyarkin	0
+I have tried to improve the information density of the paper from being to verbiose.	2	1	On page 5 the authors claim the main form of validation of the CIRA approach is a case study.			admsci5030125_boyarkin	0
+I have added the results of the stakeholder analysis to improve the communication our view of the PRP.	2	1	You stated 9 risks in table 3 while you mentioned eight in the title of that table as well as on page 9 in the beginning of 4.2.1 section.			admsci5030125_boyarkin	0
+I have tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	As an editor I would think twice if a young scientist whom I don't know offers her./himself as a reviewer and I would certainly ask for recommendations.			admsci5030125_boyarkin	0
+I have elaborated on all the areas you commented, and clarified the CIRA methodology and hopefully my contribution.	2	1	There is of course a demand for reliable reviewers but an editor is invited by a publisher because he knows the field and the players.			admsci5030125_boyarkin	0
+Thank you for comments on the grammar issues, these are now fixed. Abbrevation issues should be fixed.	2	1	Kind regards, Gaute Wangen Second Round of Evaluation Round 2: Despite the article even became longer, the line of thought is now clear.			admsci5030125_boyarkin	0
+I have also tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	4) The abbreviated version of RCA has been used once before stating it with its extended version for the first time.			admsci5030125_boyarkin	0
+I have included all your suggested changes together with a major spellcheck.	4	1	You have some examples of what has been done in the area of peer review process and the risk analysis of such a process.			admsci5030125_boyarkin	0
+I have tried to address your principle concern by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	Best regards, Gaute Wangen Round 2: It is great to see that most of the comments have been considered and there were actions to fix those in the revised version of the paper.			admsci5030125_makarova	0
+I have tried to improve the information density of the paper from being to verbiose.	2	1	Possible risks in peer review are many more than the ones listed in Table 3.			admsci5030125_makarova	0
+I have added the results of the stakeholder analysis to improve the communication our view of the PRP.	2	1	Kind regards, Gaute Wangen Round 1: and Author Response The topic of CIRA is interesting; the choice of the PRP as a case is certainly not bad.			admsci5030125_makarova	0
+I have tried to address your other concerns by improving the scope of the work, revising the method section, results, discussion and conclusion.	2	1	You did cite a reference in this regard, but you may need to briefly explain it.			admsci5030125_makarova	0
+I have elaborated on all the areas you commented, and clarified the CIRA methodology and hopefully my contribution.	2	1	2) There are some typos, punctuations and grammatical errors in the paper, such as: Some grammatical errors such as the following sentence in the abstract: “This work will mainly benefit practitioners… by providing a new approach to risk manage human factors…”.			admsci5030125_makarova	0
+I have included all your suggested changes together with a major spellcheck.	4	1	al.” rather than “et al.”, page 3 “Editor” rather than “editor” “E.g.” rather than “e.g.” Some commas are needed; e.g.			admsci5030125_makarova	0
+Agree that it should not be an issue. Therefore, I recalibrated the piece to note that co-operatives are not third sector organizations, but instead have similar features that could allow the institutional analyst fertile ground for innovations in governance.	2	1	Round 1: Author Response to Reviewer 1 I copied and pasted the reviewer commentary, below.			admsci5030148_devetyaro	0
+I agree that this is not a case study; I made the mistake in haste. I have therefore changed the paper from a case study to a discussion paper. Additionally, I note in the paper that I perform an archival analysis of media accounts in an effort to demonstrate how the utilization of the ODPs for analysizing secondary data may better position the analyst to perform rigorous, interogative fieldwork.	2	1	Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			admsci5030148_devetyaro	0
+See the DISCUSSION and CONCLUSION sections, where I have added additional analyses.	2	1	The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			admsci5030148_devetyaro	0
+Corrected and accounted for.	2	1	Nonprofit and Voluntary Sector Quarterly 41(3): 431-451.			admsci5030148_devetyaro	0
+I do not believe this is a necessary reference as I cite the International Co-operative Alliance principles, and the Rochdale Principles are somewhat commonly understood.	2	1	Round 2: Author Response to Reviewer 1 Thank you so much for your efforts.			admsci5030148_devetyaro	0
+Accounted for.	2	1	The writing is good, there is a sound development of the theory, and some well drawn conclusions.			admsci5030148_devetyaro	0
+Addressed in the text.	2	1	This is unclear in the paper - a methodology chapter/discussion is needed.			admsci5030148_devetyaro	0
+Corrected.	2	1	In addition, the developed model (Figure 1) is not new.			admsci5030148_devetyaro	0
+I have changed the headings for these sections to better reflect the content. They read as follows: 2.	4	1	Minor issues (details really): - Wrong reference number on page 4, line 14: should be 15.			admsci5030148_devetyaro	0
+This is general knowledge, thereby not necessitating a reference. If the reviewers disagree, I would add a reference.	4	1	The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation.			admsci5030148_devetyaro	0
+I have added commentary about moral hazard and information asymetries.	4	1	"COMMENT #6: Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"			admsci5030148_devetyaro	0
+Agreed. Agreed. I have added additional elaboration.	4	1	To address these subtlies see: Turnbull (2002) A New Way to Govern Cornforth (2004) The Governance of Associations and Mutuals: A Paradox Perspective Ridley-Duff (2007) Communitarian Perspectives on Social Enterprise			admsci5030148_devetyaro	0
+Commentary added, nothing the need for additiona interogation.	4	1	To address these subtlies see: Turnbull (2002) A New Way to Govern Cornforth (2004) The Governance of Associations and Mutuals: A Paradox Perspective Ridley-Duff (2007) Communitarian Perspectives on Social Enterprise			admsci5030148_devetyaro	0
+Commentary added for further clarity.	4	1	Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.			admsci5030148_devetyaro	0
+"I agree with your points about the over-emphasis on principle-agent types of governance arrangements. The Cornforth (2004) article reoriented my thinking (thank you). I recognized that it was not ""P-A"" I was critiquing, but the unitarist tradition. That approach helped with definitional rigor"	2	1	For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors.			admsci5030148_devetyaro	0
+"I believe the recalibration of the core critique from ""P-A"" to unitarism addresses the commentary, here."	2	1	The style is more journalistic and free opinion than rigourous research.			admsci5030148_devetyaro	0
+"A further recalibration. I adusted the commentary to address ""pluralistic"" governance as opposed to stakeholder governance."	2	1	Thank you very much for your helpful commentary!			admsci5030148_devetyaro	0
+I editted the paper for consistenct, in line with your commentary.	2	1	- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			admsci5030148_devetyaro	0
+The definition used is from the official representative body, the International Co-operative Alliance.	2	1	The author only focuses on the application of Ostrom design principles to nonprofit governance, thereby neglecting the solid theoretical foundation that is absolutely necessary to build a more extensive governance framework for third sector organizations.			admsci5030148_devetyaro	0
+Acknowledged. First, the paper is no longer listed as a case study, but as a discussion piece. Second, I note in the paper that I am not making the claim that the co-operative model is part of the third sector.	2	1	Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.			admsci5030148_devetyaro	0
+"Acknowledged. The ""Literature Review"" section now falls under a more accurate heading of ""Introducing the Ostrom Design Principles."""	2	1	THEORISATION Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.			admsci5030148_devetyaro	0
+I believe that this then addresses the issues that fall in line with your critiques about the theoretical foundation.	2	1	COMMENT #4: The comments added in the beginning of Chapter 3 on methodology are important and improves the text a lot.			admsci5030148_devetyaro	0
+As per your suggestions, I did focus on the co-operative model I believe that Cornforth’s paper (2004) might be a good starting point.	2	1	I still have doubts about the basis for conclusion and generalization of findings.			admsci5030148_devetyaro	0
+Very helpful! Thank you!	2	1	A sectoral comparison of wage levels and wage inequality in human services industries.			admsci5030148_devetyaro	0
+Note that the paper no longer claims this to be a case study, and instead positions itself as a discussion paper, performing a secondary archival analysis of media accounts.	2	1	"COMMENT #6: Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"			admsci5030148_devetyaro	0
+Thank you.	2	1	I still have doubts about the basis for conclusion and generalization of findings.			admsci5030148_devetyaro	0
+I agree with your critique. I have adjusted accordingly. I did this by separating the co-operative model from the third sector, and instead noted that there exists commonalities which allows for comparative learning and innovations in governance.	2	1	Turning to the good things: I do like the introduction and idea of analysing by using Ostrom's design principles, and I do find the CEC case interesting - as far as the given information goes.			admsci5030148_devetyaro	0
+Note that throughout the document I incorporate Coule's unitary-pluralist dimensions.	2	1	Second, the board-manager relationship is only a small part of the research on nonprofit boards, which in turn is the major focus of nonprofit governance research (Cornforth, 2012; Renz and Andersson, 2014).			admsci5030148_devetyaro	0
+Agreed and accounted for.	2	1	Nonprofit governance and accountability: Broadening the theoretical perspective.			admsci5030148_devetyaro	0
+Agreed. I misrepresented the analysis. It is not a case study, but instead an cursory archival analysis and exploration, utilizing secondary sources.	2	1	However, there are subtleties in the literature that need considering, and also the methodology (which is extremely weak).			admsci5030148_devetyaro	0
+Agreed. The Analysis, Discussiom, and Conclusion sections are more fleshed out.	2	1	Second, the board-manager relationship is only a small part of the research on nonprofit boards, which in turn is the major focus of nonprofit governance research (Cornforth, 2012; Renz and Andersson, 2014).			admsci5030148_devetyaro	0
+Agreed, and accounted for.	2	1	Thank you very much for your helpful commentary!			admsci5030148_devetyaro	0
+I hope that the revisions in the abstract an introduction better clarify the purpose.	2	1	THEORISATION Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.			admsci5030148_devetyaro	0
+Agreed. I accounted for this throughout the document. Terminology is used in a definitionally consistent manner.	2	1	Your comments have made this paper significantly better.			admsci5030148_devetyaro	0
+Note that I adjusted the paper, and recalibrated it from a case study, to a discussion paper that utilizes an archival analysis of secondary sources. I do this in order to demonstrate how the Ostrom Design Principles may be used by the researcher to better contextualize the subject being studied so they may enter the field prepared to deeply interrogate.	2	1	Cornforth (2004) has already analyzed the governance of co-operatives from multiple theoretical R5 perspectives.			admsci5030148_devetyaro	0
+Noted and accounted for throughout the document.	2	1	I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).			admsci5030148_devetyaro	0
+Acknowledged. I cleaned and honed the Introduction, and added a more robust Conclusion.	2	1	These apply in both cases because funding is sought from wealthy providers with the power to frame laws that require their investees to use the money for the purposes stimpulated by them.			admsci5030148_devetyaro	0
+Therefore, I recalibrated the piece to note that co-operatives are not third sector organizations, but instead have similar features that could allow the institutional analyst fertile ground for innovations in governance.	2	1	It is very similar to the nonprofit governance model of Van Puyvelde et al.			admsci5030148_makarova	0
+I have therefore changed the paper from a case study to a discussion paper. Additionally, I note in the paper that I perform an archival analysis of media accounts in an effort to demonstrate how the utilization of the ODPs for analysizing secondary data may better position the analyst to perform rigorous, interogative fieldwork.	2	1	These apply in both cases because funding is sought from wealthy providers with the power to frame laws that require their investees to use the money for the purposes stimpulated by them.			admsci5030148_makarova	0
+See the DISCUSSION and CONCLUSION sections, where I have added additional analyses.	2	1	It is good to see the author is open for suggestions from reviewers.			admsci5030148_makarova	0
+Corrected and accounted for.	2	1	ANALYSIS The framework for this section exists but feels a bit superficial and under-developed.			admsci5030148_makarova	0
+I do not believe this is a necessary reference as I cite the International Co-operative Alliance principles, and the Rochdale Principles are somewhat commonly understood.	2	1	COMMENT #3: On top of page 11 there is still no examples of adversarial effects removed by co-operative.			admsci5030148_makarova	0
+Accounted for.	2	1	The writing is good, there is a sound development of the theory, and some well drawn conclusions.			admsci5030148_makarova	0
+Addressed in the text.	2	1	This is unclear in the paper - a methodology chapter/discussion is needed.			admsci5030148_makarova	0
+Corrected.	2	1	The text itself is now almost finished, I only have a few minor comments: - The literature part is improved by changing the headline, putting it in the right setting.			admsci5030148_makarova	0
+I have changed the headings for these sections to better reflect the content.	4	1	Round 1: and Author Response The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			admsci5030148_makarova	0
+This is general knowledge, thereby not necessitating a reference. If the reviewers disagree, I would add a reference.	4	1	It is very similar to the nonprofit governance model of Van Puyvelde et al.			admsci5030148_makarova	0
+I have added commentary about moral hazard and information asymetries.	4	1	Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.			admsci5030148_makarova	0
+Agreed. Agreed. I have added additional elaboration.	4	1	Voluntas 22:566–586 R6 Round 1: Author Response to Reviewer 2 I copied and pasted the reviewer commentary, below.			admsci5030148_makarova	0
+Commentary added, nothing the need for additiona interogation.	4	1	Public enterprises, third sector cooperatives are all mixedup.			admsci5030148_makarova	0
+Commentary added for further clarity.	4	1	To address these subtlies see: Turnbull (2002) A New Way to Govern Cornforth (2004) The Governance of Associations and Mutuals: A Paradox Perspective Ridley-Duff (2007) Communitarian Perspectives on Social Enterprise			admsci5030148_makarova	0
+I agree with your points about the over-emphasis on principle-agent types of governance arrangements.	2	1	I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).			admsci5030148_makarova	0
+"I believe the recalibration of the core critique from ""P-A"" to unitarism addresses the commentary, here."	2	1	Some well grounded comments on their use in other governance research are merited right at the end.			admsci5030148_makarova	0
+"A further recalibration. I adusted the commentary to address ""pluralistic"" governance as opposed to stakeholder governance."	2	1	In addition, the developed model (Figure 1) is not new.			admsci5030148_makarova	0
+I editted the paper for consistenct, in line with your commentary.	2	1	"R14 COMMENT #5: Conclusion: middle of page 16: ""we cannot know the motivations behind.."" Have you tried to talk to them?"			admsci5030148_makarova	0
+The definition used is from the official representative body, the International Co-operative Alliance.	2	1	THEORISATION Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.			admsci5030148_makarova	0
+As per your suggestions, I did focus on the co-operative model I believe that Cornforth’s paper (2004) might be a good starting point.	2	1	Several stakeholder governance frameworks for third sector organizations have already been developed in the literature (Young, 2011; Van Puyvelde et al., 2012; Wellens and Jegers, 2014).			admsci5030148_makarova	0
+I agree with your critique. I have adjusted accordingly.	2	1	I did this by separating the co-operative model from the third sector, and instead noted that there exists commonalities which allows for comparative learning and innovations in governance.			admsci5030148_makarova	0
+Note that throughout the document I incorporate Coule's unitary-pluralist dimensions.	2	1	Nonprofit and Voluntary Sector Quarterly 41(3): 431-451.			admsci5030148_makarova	0
+Agreed and accounted for.	2	1	Annals of Public and Cooperative Economics, 75(1), 11-32.			admsci5030148_makarova	0
+Agreed. I misrepresented the analysis. It is not a case study, but instead an cursory archival analysis and exploration, utilizing secondary sources.	2	1	It is very similar to the nonprofit governance model of Van Puyvelde et al.			admsci5030148_makarova	0
+Agreed. The Analysis, Discussiom, and Conclusion sections are more fleshed out.	2	1	Thanks so much for your thoughtful commentary!			admsci5030148_makarova	0
+Agreed, and accounted for.	2	1	Instead of being vague and using a mixed terminology, the manuscript should clearly distinguish which nonprofit type is the subject of the paper, and be very careful in generalizing the findings of this study to third sector governance, which mistakenly happens in the conclusion (p. 15).			admsci5030148_makarova	0
+I hope that the revisions in the abstract an introduction better clarify the purpose.	2	1	Instead of being vague and using a mixed terminology, the manuscript should clearly distinguish which nonprofit type is the subject of the paper, and be very careful in generalizing the findings of this study to third sector governance, which mistakenly happens in the conclusion (p. 15).			admsci5030148_makarova	0
+Agreed. I accounted for this throughout the document. Terminology is used in a definitionally consistent manner.	2	1	The writing is good, there is a sound development of the theory, and some well drawn conclusions.			admsci5030148_makarova	0
+Note that I adjusted the paper, and recalibrated it from a case study, to a discussion paper that utilizes an archival analysis of secondary sources.	2	1	- The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?			admsci5030148_makarova	0
+Noted and accounted for throughout the document.	2	1	The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			admsci5030148_makarova	0
+Acknowledged. Acknowledged. I cleaned and honed the Introduction, and added a more robust Conclusion.	2	1	THEORISATION Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.			admsci5030148_makarova	0
+We fundamentally reorganized our manuscript so that its story and structure might be much clearer in the revision. In Section 2.2 we explicitly presented three hypotheses that are tested and discussed in later sections using information on six cluster cases. Section 5 also was reorganized so that the relationship of local cluster management to basic conditions and to national cluster polity, respectively, could be tested for each country using some cases of clusters and cluster management.	2	1	It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number.			admsci5040213_boyarkin	0
+We revised and simplified Figure 1 by deleting cluster performance and its relationships with other factors because in fact we do not address them in our manuscript.	2	1	KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			admsci5040213_boyarkin	0
+We read and cited in our manuscript the suggested and other references that correspond to [25], [31], [32], [33], [34], [35] and [36] (especially in Section 2.2) in revising our conceptual framework.	2	1	Freeman, C., 1988: Japan: A new National System of Innovation?			admsci5040213_boyarkin	0
+This manuscript addresses the relationship of national cluster policy and its local implementation, and not the policy organization within the government. Therefore, we slightly changed the titles of the manuscript and Section 5, and revised related terms throughout the manuscript, using the terms such as “local implementation” or “local management”.	2	1	; Soskice, D. (eds): Varieties of capitalism: The institutional foundations of comparative advantage.			admsci5040213_boyarkin	0
+The revised version went through a professional, native English editing.	4	1	Freeman, C., 1995: The National System of Innovation in Historical Perspective.			admsci5040213_boyarkin	0
+Indeed, it is not a big surprise that the characteristics of national cluster policies and their local implementation are consistent, but still it is our (small but significant) contribution to concretely show the consistency in international comparison with original interviews of cluster managers. It is beyond the scope of our manuscript to answer the question as to which model is better for the cluster performance, so we leave it as a future research agenda.	4	1	The authors should include a discussion on how these R4 countries differ e.g.			admsci5040213_boyarkin	0
+We revised the concept and its explanation, and added some references in Section 2.2 and in Introduction. For more details, please see below.	2	1	But the analysis of the data is weak, and the paper remains largely descriptive.			admsci5040213_boyarkin	0
+In this manuscript, we do not intend to demonstrate direct performance effect of cluster conditions, cluster policy, and cluster management. This is beyond the scope of our paper. It would be an important research topic to challenge the “technocratic faith”, but we cut the discussion about the factors of cluster performance from the text and Figure 1 in order to avoid any misunderstanding. Instead, we added a detailed discussion about “initial conditions” (now “basic conditions”) of clusters in Section 2.2 and 3.1 (a new section).	2	1	For instance, what does it mean if a regional cluster organisation is pre-dating the national policy programme it is supposed to implement, as evident from some of the cases?			admsci5040213_boyarkin	0
+In page 4 below in Section 2.2, we added a brief explanation of the “initial” (now “basic”) conditions of clusters. There we suggested that they comprise various regional characteristics and that also R8 scientific or industrial focus may differ across clusters within life science or biotechnology. In the new Section 3.1, we explained the differences of basic conditions in more detail.	2	1	In the following, we will reply to each of your comments, using blue letters for our responses.			admsci5040213_boyarkin	0
+It is true that in some cases local cluster organization had been established and had started its activity before the focal cluster policy started. Because of limited information, we could not sufficiently consider cluster and policy dynamics over time, but referred to the lack of dynamic approach in our study in the conclusion.	2	1	Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp.			admsci5040213_boyarkin	0
+* We recognized that we did not concretely explain the initial (we changed the term to “basic” in the revision) conditions of clusters in the previous version. Moreover, we recognize that it is important to discuss in detail how these countries differ regarding basic conditions and why the national cluster policies differ across them. Therefore, we fundamentally changed the structure of our manuscript and set up a new Section 3.1 to discuss these issues in detail. Here, we cited the suggested book chapter and paper, but mainly used statistical evidence from OECD Science, Technology and Industry Scoreboard on (different) innovation systems to characterize basic conditions of clusters and to distinguish between private- and public-driven clusters. We summarized the discussion in Table 1 (new).	2	1	In the following, we will reply to each of your comments, using blue letters for our responses.			admsci5040213_boyarkin	0
+Maybe “intellectual cluster” is a specific jargon in the Japanese policy. Following your suggestion, and according to the EU reports we cited in our manuscript, we deleted the adjective “intellectual” for clusters and just use “clusters”, because it does neither affect our concept nor findings.	2	1	In sum, I recommend that the paper be reconsidered after a major revision.			admsci5040213_boyarkin	0
+We did not include USA in the international comparison because, as you correctly suggested, there is no comparable national cluster policy there. However, we use USA in Table 1 on basic conditions as the baseline reference for the three countries.	2	1	This should be backed up with evidence (data) and preferably rephrased, such as ‘recognised R13 research universities’, or ‘elite’ with reference to the German federal government’s programme of university excellence (Exzellenzinitiative).			admsci5040213_boyarkin	0
+We recognize that the term “representative” is misleading. We replaced it with “outstanding” in the revised manuscript.	2	1	• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			admsci5040213_boyarkin	0
+We added detailed information about the number and types of interview partners and interview time on page 9 at the end of Section 2.3 (Research Methodology).	2	1	As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing.			admsci5040213_boyarkin	0
+* We did not interview the officers in charge of cluster policy in the government because information on national cluster policy was sufficiently available from cluster managers and second sources including ministry’s websites and because we are more interested in the relationship of cluster policy with basic conditions and local management than in the cluster policy itself.	2	1	This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation.			admsci5040213_boyarkin	0
+With 3 references, we discussed in the (newly inserted) third paragraph of Section 2.3 the advantages of comparative case studies across countries focusing on biotechnology.	2	1	How can these be ‘representative’ for whatever population of biotech clusters in these countries?			admsci5040213_boyarkin	0
+The classification between top-down and bottom-up clusters is misleading. In the conceptual framework we distinguish between public-driven and private-driven clusters, so that in the revised version we focused on this type of distinction and cut all descriptions on top-down and bottom-up clusters.	2	1	Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_boyarkin	0
+Referring to Kiese (2013) and Sternberg et al. (2010), but also other papers such as Crespy et al. (2007) for France, we added that, especially for Germany, we should not underestimate the roles of states and local authorities in the cluster policy. Finally, in the conclusion, we stressed the lack of multilevel governance approach as a limitation, which should be more explicitly considered in future research.	2	1	When explaining the choice of regional cases, the term ‘representative’ appears misleading.			admsci5040213_boyarkin	0
+We recognized that this section has several problems, but only this section describes and compares local cluster management based on our own interviews, so that this is the core part of our manuscript. Therefore, we fundamentally changed this section in the following way. First, we have some reference in the revised version, whereas this section is mainly based on our interviews. Second, at the beginning of this section, we explained the relevance of this section in our concept, especially with regard to our (new) hypotheses. Third, more importantly, we reorganized this section to a comparison of cluster management across three countries, focusing on the selection procedure of joint R&D R11 projects (and cutting other information such as monitoring process and support programs). In this way, we made it clear what we do in this section: to check the consistency of local cluster management (regarding project selection) with the cluster’s basic conditions and the national policy. We believe that we could clarify the relevance of the content of this section with the preceding part and the conclusion of this manuscript.	2	1	Even when the industry (biotechnology) is held constant, clusters differ in a number of ways, esp.			admsci5040213_boyarkin	0
+Cluster Organization’s governance and financing, which was already described in another section in the previous version, is now explained even earlier, in Section 3.2.	2	1	Round 2: Author Response to Reviewer 1 Thank you for your comments.			admsci5040213_boyarkin	0
+We cut the most part of the discussion in the conclusion (which was mostly redundant with the contents of preceding sections), and added policy implication and some limitations (lack of multilevel governance and dynamic perspectives, possible selection bias of target clusters, and the descriptive and qualitative nature of the study) combined with future research perspectives.	2	1	Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_boyarkin	0
+We cited this paper in footnote 4 in page 3.	2	1	However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified.			admsci5040213_boyarkin	0
+We agree with the suggestion and changed “few” to “no”. We also checked all the other terms “few” in the manuscript and treated them in the suggested way.	2	1	It looks as if one interview was done with the cluster manager in the six regions, but the authors also claim to have interviewed ‘the presidents of cluster firms’ (p. 8, line 20) without indicating the number.			admsci5040213_boyarkin	0
+We changed the sentence (now page 6, line 22) in the following way: “may enhance the incentives of member firms to succeed and partially prevent moral hazard and crowding out”.	2	1	• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			admsci5040213_boyarkin	0
+Based on the argument in Section 2.2, we presented three hypotheses in page 7, which are repeatedly addressed in later sections and the conclusion.	2	1	• Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.			admsci5040213_boyarkin	0
+We found no evidence on policy learning between Germany and France.	2	1	• Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’.			admsci5040213_boyarkin	0
+We changed the phrase to “a series of national cluster policy” (now page 18, line 5) at the beginning of Section 4.2.	2	1	Taking these up in the empirical discussion could help the reader’s orientation.			admsci5040213_boyarkin	0
+We noticed in footnote 18 in page 18 that some cluster regions may have applied for this program twice or three times after failing in the previous competition.	2	1	It seems that the most prominent or successful cases have been selected, at least for Germany and France.			admsci5040213_boyarkin	0
+We cited these studies in Introduction and in Section 4.3 (page 18, footnotes 20 and 21).	2	1	in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up.			admsci5040213_boyarkin	0
+We cut the previous Table 2 (on each target cluster and cluster management) in the revision and also the related sentence.	2	1	The authors might find the following major and minor comments helpful when revising their paper.			admsci5040213_boyarkin	0
+We agree with you. However, according to the MEXT’s definition, clusters need not involve local high-tech ventures. R&D collaboration between local public research institute and private firms outside of the cluster area is also an important part of a cluster for MEXT. Regarding the aim of local authorities, which includes the development of a cluster around a research organization or hospital, indeed cluster policy has not produced any outcomes yet.	2	1	in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up.			admsci5040213_boyarkin	0
+We used “prominent” and “excellent” instead of “famous”.	2	1	As Kiese (2009) showed for three European Countries including Germany and France, these differences leave an imprint on the design and implementation of national cluster policies.			admsci5040213_boyarkin	0
+We cut this sentence.	2	1	All remarks have been considered, most have been fully incorporated and the paper has gained substantially from revision.			admsci5040213_boyarkin	0
+We cited this paper in footnote 14 in Section 3.2. (3) to mention that Trinational BioValley has hardly had any impact in terms of intensifying cross-border interaction.	2	1	The authors seem to assume that cluster policies are either bottom-up or top-down, without any shades of grey in between.			admsci5040213_boyarkin	0
+We changed the layout of the tables enlarging the font.	2	1	In: Environment and Planning C, 28(6): 1063-1082.			admsci5040213_boyarkin	0
+In the revision, we write the funding amount for Japan in Yen, but give the converged amount in Euro in parentheses. We use the most recent exchange rate for convenience, and explain it where it is first mentioned (note to Table 1 in page 16 and footnote 17 in page 17).	2	1	R7 LONGHI, C.; ROCHHIA, S., 2013: Cluster policy for innovation and competitiveness: Lessons from the French experience.			admsci5040213_boyarkin	0
+We added a discussion on the national systems of innovation in the first two paragraphs of Section 3.1 (p. 9) referring to the suggested seminal works. Moreover, in the remaining part of this section (pp. 10-11), we added some sentences with supporting references on the underlying institutional differences.	4	1	Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique.			admsci5040213_boyarkin	0
+At the beginning of Section 3.2, as well as at the end of the description of each cluster, we mitigated the dichotomist nature in the typology of cluster development by some qualification. For example, we changed the formulation from “characterized as public-driven clusters” to “characterized as more public-driven than private-driven clusters”.	4	1	The paper thus rests on the untested presumption that cluster management and national policies actually impact the performance of clusters.			admsci5040213_boyarkin	0
+We added a new reference on the development in Japan (Okubo and Tomiura 2010) but deleted the sentence about France because we could not show a reference in English.	4	1	The language the authors use is here a bit imprecise - they seem to be talking about the management of cluster initiatives, not about the management of the policies themselves (which is more a question of how the program oversight is organized within government; also an interesting but different question).			admsci5040213_boyarkin	0
+According to this suggestion, we dropped footnote 14 with the related reference.	4	1	When outlining their research methodology, the authors should state precisely how many interviews they conducted.			admsci5040213_boyarkin	0
+We added the information sources under the table.	4	1	• Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references.			admsci5040213_boyarkin	0
+We replaced “policy” by “programs” in this sentence.	4	1	Remaining Comments Although the new section 3.1 helps situating the cases within their respective national contexts, it’s focus is on performance indicators.			admsci5040213_boyarkin	0
+In Conclusion, we presented again three hypotheses in a new paragraph in Conclusion after the first paragraph (p. 22). Then, in the following paragraph (p. 23) we added a discussion on how differences in cluster policies and cluster management are linked to those in national systems of innovation, with three sentences.	4	1	; Soete, L. (eds): Technical Change and Economic Theory.			admsci5040213_boyarkin	0
+We fundamentally reorganized our manuscript so that its story and structure might be much clearer in the revision. In Section 2.2 we explicitly presented three hypotheses that are tested and discussed in later sections using information on six cluster cases. Section 5 also was reorganized so that the relationship of local cluster management to basic conditions and to national cluster polity, respectively, could be tested for each country using some cases of clusters and cluster management.	2	1	KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			admsci5040213_devetyaro	0
+We revised and simplified Figure 1 by deleting cluster performance and its relationships with other factors because in fact we do not address them in our manuscript.	2	1	Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_devetyaro	0
+We read and cited in our manuscript the suggested and other references that correspond to [25], [31], [32], [33], [34], [35] and [36] (especially in Section 2.2) in revising our conceptual framework.	2	1	It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.			admsci5040213_devetyaro	0
+This manuscript addresses the relationship of national cluster policy and its local implementation, and not the policy organization within the government. Therefore, we slightly changed the titles of the manuscript and Section 5, and revised related terms throughout the manuscript, using the terms such as “local implementation” or “local management”.	2	1	Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.			admsci5040213_devetyaro	0
+Indeed, it is not a big surprise that the characteristics of national cluster policies and their local implementation are consistent, but still it is our (small but significant) contribution to concretely show the consistency in international comparison with original interviews of cluster managers. It is beyond the scope of our manuscript to answer the question as to which model is better for the cluster performance, so we leave it as a future research agenda.	4	1	• Methodology: The choice of clusters is generally well explained on pp.			admsci5040213_devetyaro	0
+We revised the concept and its explanation, and added some references in Section 2.2 and in Introduction. For more details, please see below.	2	1	Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.			admsci5040213_devetyaro	0
+In this manuscript, we do not intend to demonstrate direct performance effect of cluster conditions, cluster policy, and cluster management. This is beyond the scope of our paper. It would be an important research topic to challenge the “technocratic faith”, but we cut the discussion about the factors of cluster performance from the text and Figure 1 in order to avoid any misunderstanding. Instead, we added a detailed discussion about “initial conditions” (now “basic conditions”) of clusters in Section 2.2 and 3.1 (a new section).	2	1	This reformulation is incorrect as the Koschatzky (2000) paper pre-dated the BioValley initiative and does not look at INTERREG either.			admsci5040213_devetyaro	0
+In page 4 below in Section 2.2, we added a brief explanation of the “initial” (now “basic”) conditions of clusters. There we suggested that they comprise various regional characteristics and that also R8 scientific or industrial focus may differ across clusters within life science or biotechnology.	2	1	• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			admsci5040213_devetyaro	0
+It is true that in some cases local cluster organization had been established and had started its activity before the focal cluster policy started. Because of limited information, we could not sufficiently consider cluster and policy dynamics over time, but referred to the lack of dynamic approach in our study in the conclusion.	2	1	• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			admsci5040213_devetyaro	0
+We recognized that we did not concretely explain the initial (we changed the term to “basic” in the revision) conditions of clusters in the previous version. Moreover, we recognize that it is important to discuss in detail how these countries differ regarding basic conditions and why the national cluster policies differ across them. Therefore, we fundamentally changed the structure of our manuscript and set up a new Section 3.1 to discuss these issues in detail. Here, we cited the suggested book chapter and paper, but mainly used statistical evidence from OECD Science, Technology and Industry Scoreboard on (different) innovation systems to characterize basic conditions of clusters and to distinguish between private- and public-driven clusters. We summarized the discussion in Table 1 (new).	2	1	the interface between national cluster policies and cluster management on the ground, and thus makes an original contribution to our understanding of cluster policies and their implementation.			admsci5040213_devetyaro	0
+Maybe “intellectual cluster” is a specific jargon in the Japanese policy. Following your suggestion, and according to the EU reports we cited in our manuscript, we deleted the adjective “intellectual” for clusters and just use “clusters”, because it does neither affect our concept nor findings.	2	1	• Page 15, line 24: Here and elsewhere, universities are attributed with the term ‘famous’.			admsci5040213_devetyaro	0
+We did not include USA in the international comparison because, as you correctly suggested, there is no comparable national cluster policy there. However, we use USA in Table 1 on basic conditions as the baseline reference for the three countries.	2	1	In the new Section 3.1, we explained the differences of basic conditions in more detail.			admsci5040213_devetyaro	0
+We recognize that the term “representative” is misleading. We replaced it with “outstanding” in the revised manuscript.	2	1	Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e.			admsci5040213_devetyaro	0
+We added detailed information about the number and types of interview partners and interview time on page 9 at the end of Section 2.3 (Research Methodology).	2	1	• Page 4, line 23: ‘few studies’ – If you mean ‘few’, please state the references.			admsci5040213_devetyaro	0
+We did not interview the officers in charge of cluster policy in the government because information on national cluster policy was sufficiently available from cluster managers and second sources including ministry’s websites and because we are more interested in the relationship of cluster policy with basic conditions and local management than in the cluster policy itself.	2	1	The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies.			admsci5040213_devetyaro	0
+With 3 references, we discussed in the (newly inserted) third paragraph of Section 2.3 the advantages of comparative case studies across countries focusing on biotechnology.	2	1	Furthermore, the state government of Bavaria may be a regional government, but it is certainly not a local authority (like the city of Munich; page 15, line 29) in Germany’s system of multilevel governance.			admsci5040213_devetyaro	0
+The classification between top-down and bottom-up clusters is misleading. In the conceptual framework we distinguish between public-driven and private-driven clusters, so that in the revised version we focused on this type of distinction and cut all descriptions on top-down and bottom-up clusters.	2	1	When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!			admsci5040213_devetyaro	0
+Referring to Kiese (2013) and Sternberg et al. (2010), but also other papers such as Crespy et al. (2007) for France, we added that, especially for Germany, we should not underestimate the roles of states and local authorities in the cluster policy. Finally, in the conclusion, we stressed the lack of multilevel governance approach as a limitation, which should be more explicitly considered in future research.	2	1	For the German cases, the authors may have underestimated public agency, since they did not investigate the role of state governments (Bavaria, Baden-Württemberg) within the country’s system of multilevel governance, which is key to understand cluster policies in Germany (cf.			admsci5040213_devetyaro	0
+We recognized that this section has several problems, but only this section describes and compares local cluster management based on our own interviews, so that this is the core part of our manuscript. Therefore, we fundamentally changed this section in the following way. First, we have some reference in the revised version, whereas this section is mainly based on our interviews. Second, at the beginning of this section, we explained the relevance of this section in our concept, especially with regard to our (new) hypotheses. Third, more importantly, we reorganized this section to a comparison of cluster management across three countries, focusing on the selection procedure of joint R&D R11 projects (and cutting other information such as monitoring process and support programs). In this way, we made it clear what we do in this section: to check the consistency of local cluster management (regarding project selection) with the cluster’s basic conditions and the national policy. We believe that we could clarify the relevance of the content of this section with the preceding part and the conclusion of this manuscript.	2	1	I would still like to see a qualification stating that R16 private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.			admsci5040213_devetyaro	0
+Cluster Organization’s governance and financing, which was already described in another section in the previous version, is now explained even earlier, in Section 3.2.	2	1	Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_devetyaro	0
+We cut the most part of the discussion in the conclusion (which was mostly redundant with the contents of preceding sections), and added policy implication and some limitations (lack of multilevel governance and dynamic perspectives, possible selection bias of target clusters, and the descriptive and qualitative nature of the study) combined with future research perspectives.	2	1	In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.			admsci5040213_devetyaro	0
+We cited this paper in footnote 4 in page 3.	2	1	Compared to the Japanese cases, the French and German cases are clearly more bottom-up, but they still display clear – if not dominant – elements of top-down governance and public agency.			admsci5040213_devetyaro	0
+We agree with the suggestion and changed “few” to “no”. We also checked all the other terms “few” in the manuscript and treated them in the suggested way.	2	1	We tried to consider the comments and suggestions as far as possible in revising our manuscript.			admsci5040213_devetyaro	0
+We changed the sentence (now page 6, line 22) in the following way: “may enhance the incentives of member firms to succeed and partially prevent moral hazard and crowding out”.	2	1	There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti).			admsci5040213_devetyaro	0
+Based on the argument in Section 2.2, we presented three hypotheses in page 7, which are repeatedly addressed in later sections and the conclusion.	2	1	Freeman, C., 1988: Japan: A new National System of Innovation?			admsci5040213_devetyaro	0
+We found no evidence on policy learning between Germany and France.	2	1	P. 11: References on Japan and France should be added to the last two sentences of the first paragraph.			admsci5040213_devetyaro	0
+We changed the phrase to “a series of national cluster policy” (now page 18, line 5) at the beginning of Section 4.2.	2	1	It seems that the most prominent or successful cases have been selected, at least for Germany and France.			admsci5040213_devetyaro	0
+We noticed in footnote 18 in page 18 that some cluster regions may have applied for this program twice or three times after failing in the previous competition.	2	1	Also sound profiling of the different cluster programs and initiatives.			admsci5040213_devetyaro	0
+We cited these studies in Introduction and in Section 4.3 (page 18, footnotes 20 and 21).	2	1	• It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices.			admsci5040213_devetyaro	0
+We cut the previous Table 2 (on each target cluster and cluster management) in the revision and also the related sentence.	2	1	The performance of clusters is also affected by many exogenous influences not captured in the simple model.			admsci5040213_devetyaro	0
+We agree with you. However, according to the MEXT’s definition, clusters need not involve local high-tech ventures. R&D collaboration between local public research institute and private firms outside of the cluster area is also an important part of a cluster for MEXT. Regarding the aim of local authorities, which includes the development of a cluster around a research organization or hospital, indeed cluster policy has not produced any outcomes yet.	2	1	The conceptual framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included.			admsci5040213_devetyaro	0
+We used “prominent” and “excellent” instead of “famous”.	2	1	Specific Comments • Page 3, line 3: The authors might want to link to the literature on differentiated knowledge bases, which assigns an analytical (science-based) knowledge base to the biotech industry (cf.			admsci5040213_devetyaro	0
+We cut this sentence.	2	1	P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before.			admsci5040213_devetyaro	0
+We cited this paper in footnote 14 in Section 3.2. (3) to mention that Trinational BioValley has hardly had any impact in terms of intensifying cross-border interaction.	2	1	• The layout of the tables may be improved, especially the spacing of the text.			admsci5040213_devetyaro	0
+We changed the layout of the tables enlarging the font.	2	1	Round 1: Referee Report for Administrative Sciences Management of cluster policies for innovation: Comparative case studies of Japanese, German, and French biotechnology clusters Manuscript no.			admsci5040213_devetyaro	0
+In the revision, we write the funding amount for Japan in Yen, but give the converged amount in Euro in parentheses. We use the most recent exchange rate for convenience, and explain it where it is first mentioned (note to Table 1 in page 16 and footnote 17 in page 17).	2	1	P. 18 (section 4.1, top paragraph): I would rather prefer “a series of national cluster programmes” as part of an evolving policy.			admsci5040213_devetyaro	0
+The revised version went through a professional, native English editing. Thank you for your recommendation. We provided minor corrections to our manuscript based on the following comments. The revised version went through a professional, native English editing.	4	1	o P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably R15 taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?			admsci5040213_devetyaro	0
+We added a discussion on the national systems of innovation in the first two paragraphs of Section 3.1 (p. 9) referring to the suggested seminal works. Moreover, in the remaining part of this section (pp. 10-11), we added some sentences with supporting references on the underlying institutional differences.	4	1	• Page 10, line 28: The number of 85 applicants contains some clusters who applied more than once in the three rounds of the contest.			admsci5040213_devetyaro	0
+At the beginning of Section 3.2, as well as at the end of the description of each cluster, we mitigated the dichotomist nature in the typology of cluster development by some qualification. For example, we changed the formulation from “characterized as public-driven clusters” to “characterized as more public-driven than private-driven clusters”.	4	1	However, sections 4.3 to 4.5 leave an impression that the assessment of these cases as ‘bottom-up’ is not completely justified.			admsci5040213_devetyaro	0
+We added a new reference on the development in Japan (Okubo and Tomiura 2010) but deleted the sentence about France because we could not show a reference in English.	4	1	I would still like to see a qualification stating that R16 private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.			admsci5040213_devetyaro	0
+According to this suggestion, we dropped footnote 14 with the related reference.	4	1	Would suggest to use these case studies as a source of ideas on how policies and cluster initiative management might relate to each other; that would make this a more interesting paper.			admsci5040213_devetyaro	0
+We added the information sources under the table.	4	1	According to the definition on the top of page 4, the term denotes a R9 cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper.			admsci5040213_devetyaro	0
+We replaced “policy” by “programs” in this sentence.	4	1	• It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices.			admsci5040213_devetyaro	0
+In Conclusion, we presented again three hypotheses in a new paragraph in Conclusion after the first paragraph (p. 22). Then, in the following paragraph (p. 23) we added a discussion on how differences in cluster policies and cluster management are linked to those in national systems of innovation, with three sentences.	4	1	Broad Comments The paper fills an important research deficit by focusing on the interdependence between national cluster policy programmes and their implementation by cluster management on the ground.			admsci5040213_devetyaro	0
+We added Freud’s heritage and contemporary research e.g. Jung’s contribution to the Big 5.	2	1	Author Response Author Response File: Author Response.docx			admsci6020005_makarova	0
+We took great effort introducing above mentioned perspectives into our paper.	2	1	The recommendations were very constructive and we hope that we have addressed them all.			admsci6020005_makarova	0
+We decided to call them both gurus because David obviously is the one but our paper refers to Jennifer.	2	1	There has been some backlash to the application of brand personality to inanimate objects.			admsci6020005_makarova	0
+We added Allen & Olson and other limitations such as criticism on the conceptual, methodological and substantive level as well as confusing user profiles (e.g., upper class) with brand characteristics and weak discriminatory power.	2	1	(f) “Archetype Articulate Charisma” (heading title in Table 3): This is not clear.			admsci6020005_makarova	0
+We revised the statement in the sense that it is part and not different from it.	2	1	After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity.			admsci6020005_makarova	0
+We fully agree and deleted the claim to be Grounded Theory.	2	1	We hope that we have given full consideration to all comments.			admsci6020005_makarova	0
+We cited Roberts (2010) who, based on extensive research, found different leading archetypes depending on the product category. We added our thoughts as scope for further research in chapter 4.We added Matzler et al. (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) as research on customer-brand relationship.	2	1	Please don’t expect them to consult original sources by Jung!			admsci6020005_makarova	0
+The survey was conducted face-to-face in the classroom and students asked personally.	2	1	We also mentioned the difference between semiotic and formalist approach.			admsci6020005_makarova	0
+We added more screenshots and background on the commercials.	2	1	We also mentioned the difference between semiotic and formalist approach.			admsci6020005_makarova	0
+We added the questions that participants were asked. Everything was based on pencil and paper.	2	1	"Padgett, Dan and Michael S. Mulvey (2007), ""Differentiation Via Technology: Strategic Positioning of Services Following the Introduction of Disruptive Technology,"" Journal of Retailing, 83 (4), 375-91."			admsci6020005_makarova	0
+Changed in the term ‘manifestation.’ Findings: Who rated the three personality traits?	2	1	"For example the fact that some ""heroes"" can move across categories - e.g., Tiger Woods for Buick."			admsci6020005_makarova	0
+The students.	2	1	After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity.			admsci6020005_makarova	0
+Principal Component with Varimax rotation.	2	1	As mentioned at the onset of my review, I fully support the authors’ aim to revitalize and blend archetypal and personality-based approaches to brand identity.			admsci6020005_makarova	0
+We changed the wording. In fact, the cited Cinderalla is a prime example of blended archetype.	2	1	There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.			admsci6020005_makarova	0
+We cleaned up our description of the experimental design by adding the questions that we asked, the scale we used etc.	2	1	First of all we would like to thank our reviewer for introducing new perspectives.			admsci6020005_makarova	0
+We tried our best to address this problem, see the revised version of our paper.	2	1	A close reading of some of the cited work (i.e., Mark and Pearson 1991) recognizes that archetypes can be blended.			admsci6020005_makarova	0
+We added Freud’s heritage and contemporary research e.g. Jung’s contribution to the Big 5. We took the liberty to refer to Kant and Goethe re the origin of the term ‘archetype’ which may give it a more philosophical touch. We would have loved to substantiate the archetype concept by neurophysiological research but were not convinced about the reliability of claimed research findings so we added it under scope for further research (chapter: Implications).	2	1	There has been some backlash to the application of brand personality to inanimate objects.			admsci6020005_perova	0
+We took great effort introducing above mentioned perspectives into our paper. It stretches across the whole paper. We also mentioned the difference between semiotic and formalist approach.	2	1	This may be of concern, because the paper uses hotels as an example.			admsci6020005_perova	0
+We decided to call them both gurus because David obviously is the one but our paper refers to Jennifer.	2	1	"Scott, Linda M. (1994), ""The Bridge from Text to Mind: Adapting Reader-Response Theory to Consumer Research,"" Journal of Consumer Research, 21 (3), 461-80."			admsci6020005_perova	0
+We added Allen & Olson and other limitations such as criticism on the conceptual, methodological and substantive level as well as confusing user profiles (e.g., upper class) with brand characteristics and weak discriminatory power.	2	1	It is interesting and the work of Aaker may also be interpreted to represent images of Freedom, Social, Order and Ego.			admsci6020005_perova	0
+We revised the statement in the sense that it is part and not different from it.	2	1	"REFERENCES Allen, Douglas E and Jerry Olson (1995), ""Conceptualizing and Creating Brand Personality: A Narrative Theory Approach,"" Advances in Consumer Research, 22 (1), 392-93."			admsci6020005_perova	0
+We fully agree and deleted the claim to be Grounded Theory.	2	1	As mentioned at the onset of my review, I fully support the authors’ aim to revitalize and blend archetypal and personality-based approaches to brand identity.			admsci6020005_perova	0
+We cited Roberts (2010) who, based on extensive research, found different leading archetypes depending on the product category. We added our thoughts as scope for further research in chapter 4.We added Matzler et al. (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) as research on customer-brand relationship.	2	1	(c) The abstract states: “Aaker is seen by many as the branding guru” and refers to Aaker (1997) in the introductory paragraph.			admsci6020005_perova	0
+The survey was conducted face-to-face in the classroom and students asked personally. The two Asian hotel chains are globally not very prominent. Most Western executives stay at well-known global hotel chains (e.g., Accor, Hilton) because they have a corporate contract. Shangri-La and Banyan Tree are not that visible in Western countries. This stays in contrast to Asian executives.	2	1	Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).			admsci6020005_perova	0
+We added more screenshots and background on the commercials. Whereas the BT commercials features beautiful pictures that are sequence-wise interchangeable, the BT commercial unfolds and has it climax at the end with the wolves warming the traveler.	2	1	The recommendations were very constructive and we hope that we have addressed all.			admsci6020005_perova	0
+We added the questions that participants were asked. Everything was based on pencil and paper.	2	1	This may be of concern, because the paper uses hotels as an example.			admsci6020005_perova	0
+Changed in the term ‘manifestation.’ Findings: Who rated the three personality traits?	2	1	(f) “Archetype Articulate Charisma” (heading title in Table 3): This is not clear.			admsci6020005_perova	0
+The students. We added that part.	2	1	This is highly relevant to answering the “so what?” question that is addressed in the implications section.			admsci6020005_perova	0
+Principal Component with Varimax rotation. We added in the text.	2	1	(d) Indeed, Aaker (1997) is a highly-cited work.			admsci6020005_perova	0
+We changed the wording. In fact, the cited Cinderalla is a prime example of blended archetype.	2	1	I hope the authors find these comments helpful.			admsci6020005_perova	0
+We hope that we have given full consideration to all comments. We found them very useful and they increased the value of our paper considerably.	2	1	Personally, I agree with you – brand personality is a key facet of brand identity.			admsci6020005_perova	0
+We cleaned up our description of the experimental design by adding the questions that we asked, the scale we used etc. We included the example of David Beckham. In this context we cited Matzler et al (Personality, person-brand fit, and brand community: An investigation of individuals, brands, and brand communities. Journal Of Marketing Management) who looked into the archetype-brand-consumer match. We are delighted that our reviewer sees the link (as the authors do) between Aaker and Jung’s images of Freedom, Social, Order and Ego.	2	1	This may be of concern, because the paper uses hotels as an example.			admsci6020005_perova	0
+We tried our best to address this problem, see the revised version of our paper. However, this is an uphill task. It goes in the direction of advertising effectiveness. We would have loved to cite some reliable sources on advertising effectiveness e.g. by means of neurophysiological evidence based research but tried in vain.	2	1	"For example the fact that some ""heroes"" can move across categories - e.g., Tiger Woods for Buick."			admsci6020005_perova	0
+line 76 and 79: Sentence about two farms with financial support is deleted from the aim.	2	1	As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			agriculture12060868_makarova	0
+"Line 78: ""designed to meet behavioural needs...."" is deleted Overall, the objective now is focused on the hypotheses."	2	1	The following changes could improve the quality of the paper.			agriculture12060868_makarova	0
+l 80-83 is now deleted from the objectives 4.	2	1	There are only minor comments with very specific issues:			agriculture12060868_makarova	0
+Figure 1 must was lost from the last document, and we are so sorry for this. The figure is now included in the new document.	2	1	The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			agriculture12060868_makarova	0
+More explanation is included in the figure text of figure 2	2	1	However, it would be more elegant to have more batches.			agriculture12060868_makarova	0
+Corrections are made in the text of figure 4.	2	1	The reviewer is completely right in adressing this point.			agriculture12060868_makarova	0
+l 206, l225 Info is changed according to the reviewers request.	2	1	The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+Insted of inserting P-values in the text when reporting significant differences for the ones that are in tables, we have included more references to the stat-tables.	2	1	There are only minor comments with very specific issues:			agriculture12060868_makarova	0
+This is now included.	2	1	As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			agriculture12060868_makarova	0
+Stillborn piglets vs piglets with no milk: more details are included about this in the description of the M/M	2	1	However, it would be more elegant to have more batches.			agriculture12060868_makarova	0
+224-225: confusion about parity is corrected,	2	1	As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			agriculture12060868_makarova	0
+Figure 5 and 7. There is always effects of batches in these types of studies. This is also why we prefer to have several batches, but the main effect should not be biased by the batch effect in the present study, as the main finding are quite clear even in our small data set.	2	1	However, it would be more elegant to have more batches.			agriculture12060868_makarova	0
+mean + se is given in all tables and figures, Replicates are now included and abbreviations explained.	2	1	Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			agriculture12060868_makarova	0
+Italic for P-values in the text. Done!	2	1	Author Response Figure 1 was missed from the last MS.			agriculture12060868_makarova	0
+Figure 1 was no included by an error in the last MS. It is now inserted again.	2	1	Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			agriculture12060868_makarova	0
+Batches and number of litters are added to the figure legends.	2	1	The system we have used is to refer to the table with statistical values when reporting significant differences for the results presented in figures and tables, but for those that are presented in the text only, we also have P- and F-values in brackets.			agriculture12060868_makarova	0
+Space before and after sign is added.	2	1	This was unfortunately not possible in the present iinnovation project.			agriculture12060868_makarova	0
+A new and improved version of figure 6 is now included.	2	1	The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+We have inserted * to denote differences instead of letters.	2	1	The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+Table 4 is inserted again in the writing form.	2	1	The writing and data presenting need to be improved.			agriculture12060868_makarova	0
+The meaning of ***: this is explained.	2	1	Finally, thank you for being so patient with our manuscript			agriculture12060868_makarova	0
+Quality of figure 7 is improved.	2	1	The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+We better clarified this in the method section (L101, L105-106). 2000 is the year of the establishment of Conservation and Safety Network. In this frame, seeds were collected and maintained in long term storage conditions by Arsial in the subsequent years.	2	1	Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			agronomy12061433_makarova	0
+Between 8-10 grams (about 2000 seeds) we don’t think is a detail to insert in the manuscript.	2	1	The results are described and discussed in detail.			agronomy12061433_makarova	0
+Yes, we clarified (L116).	2	1	The results are described and discussed in detail.			agronomy12061433_makarova	0
+We added these details (L132 and L136) The results are described and discussed in detail.	2	1	We also thank you for the positive and encouraging comment.			agronomy12061433_makarova	0
+We prefer to keep the introduction in order to give a broad overview to readers	2	1	Additional corrections through the text have been done.			agronomy12061433_makarova	0
+"The cluster analysis is based on phenotypic traits, this analysis revealed specific clusters of ""Cornetto di Pontecorvo"" and similar types, although they were not clearly separated at the hierarchical level. The genotyping analysis provided more in-depth insights. We addressed this in the discussions section (see L485 and subsequent)."	2	1	Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			agronomy12061433_makarova	0
+Product is correct since refer to This crop (singular), we specify (as a food and non-food product)	2	1	See the attachment Comments for author File: Comments.pdf			agronomy12061433_makarova	0
+We agree and better positioned the table 3 after the sentence Line 385-388: the figure legend does not provide enough information on this figure #Answer:	2	1	2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			agronomy12061433_makarova	0
+Figure 4 caption has been improved	2	1	See the attachment Comments for author File: Comments.pdf			agronomy12061433_makarova	0
+We agree and modified the sentence (see L439)	2	1	line 115: Did one repetition mean one plant?line 130: How many fruits were used as a sample of representative fruits?			agronomy12061433_makarova	0
+We change referring to ddRAD seq (see L473) Conclusion:	2	1	1) The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.			agronomy12061433_makarova	0
+In order to answer to this question, we improved the conclusion (L524-529)	2	1	2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			agronomy12061433_makarova	0
+We better clarified this in the method section (L101, L105-106). 2000 is the year of the establishment of Conservation and Safety Network. In this frame, seeds were collected and maintained in long term storage conditions by Arsial in the subsequent years.	2	1	line 135: How many fruits were used as a bulk of representative fruits?			agronomy12061433_perova	0
+Between 8-10 grams (about 2000 seeds) we don’t think is a detail to insert in the manuscript.	2	1	There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			agronomy12061433_perova	0
+Yes, we clarified (L116).	2	1	This work is quite interesting and is well-written.			agronomy12061433_perova	0
+We added these details (L132 and L136)	2	1	Below point by point response to concerns Reviewer 1 The manuscript is generally well written.			agronomy12061433_perova	0
+We have carefully considered comments and thoughtful suggestions, revising the manuscript accordingly. All amendments suggested have been included. Additional corrections through the text have been done.	2	1	Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			agronomy12061433_perova	0
+We prefer to keep the introduction in order to give a broad overview to readers	2	1	2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			agronomy12061433_perova	0
+"The cluster analysis is based on phenotypic traits, this analysis revealed specific clusters of ""Cornetto di Pontecorvo"" and similar types, although they were not clearly separated at the hierarchical level. The genotyping analysis provided more in-depth insights. We addressed this in the discussions section (see L485 and subsequent)."	2	1	There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			agronomy12061433_perova	0
+Product is correct since refer to This crop (singular), we specify (as a food and non-food product)	2	1	 The results are described and discussed in detail.			agronomy12061433_perova	0
+We agree and better positioned the table 3 after the sentence	2	1	#Answer: Thanks We hope that these revisions successfully address the raised concerns, remaining available for any other requests regarding the manuscript.			agronomy12061433_perova	0
+Figure 4 caption has been improved	2	1	#Answer: Thanks We hope that these revisions successfully address the raised concerns, remaining available for any other requests regarding the manuscript.			agronomy12061433_perova	0
+We agree and modified the sentence (see L439)	2	1	Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			agronomy12061433_perova	0
+We change referring to ddRAD seq (see L473)	2	1	Below point by point response to concerns Reviewer 1 The manuscript is generally well written.			agronomy12061433_perova	0
+In order to answer to this question, we improved the conclusion (L524-529)	2	1	It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			agronomy12061433_perova	0
+The content of the two papers Reviewer 1 mentions here focussed on the impact of the pandemic on management changes in dogs and on dog owner experiences at a single time point during the most restrictive ‘lockdown’. The current paper is different to either of these, and indeed any other published paper, in that it focusses on changes in separation related behaviour over time, across two different surveys in a longitudinal cohort. The paper is about separation-related behaviour, not about the pandemic. The changes in leaving routine that occurred during the pandemic afforded the opportunity to study what impact this had on a large population of pet dogs over time, that would otherwise have been impossible to capture at such scale. The topic is unique and important in that it adds evidence for an association that has previously been supposed or observed by individual practitioners, but not demonstrated in the published literature, and certainly not on this scale.	2	1	Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+Yes, this was purposeful, it provides a summary of the aims & methods to introduce the discussion as not everyone reads papers in a linear fashion. We have not made any changes based on this statement.	2	1	“More Attention than Usual”: A Thematic Analysis of Dog Ownership Experiences in the 530 UK during the First COVID-19 Lockdown.			ani12040482_perova	0
+There is a difference between something that is considered to be “well-known” or folk-knowledge and demonstrable evidence. The sentence quoted here is not untrue, or exaggerated, however, if the Editor wishes us to explore this further, we would kindly request the Reviewer provide us with examples of publications where this has been empirically demonstrated, as we have not found any in our own searches.	2	1	I felt the discussion about changes in greeting behaviour as an early indicator was interesting and could be expanded.			ani12040482_perova	0
+Given the lack of constructive feedback in these statements, no changes have been made based on these comments.	2	1	I felt the discussion about changes in greeting behaviour as an early indicator was interesting and could be expanded.			ani12040482_perova	0
+Thank you, in line 16 and 32 we have added “when restrictions had eased” to explain why we surveyed in October 2020.	2	1	 Line 461: change “onto” to “on to” Thank you, we’ve corrected the typo.			ani12040482_perova	0
+Thank you, we have now expanded this to read: “In addition to being a welfare concern for the dog, SRBs are often unwanted or problematic for the owner and/or neighbours such as in the case of excessive vocalisation, which may lead to noise complaints, or in terms of house soiling and destruction of property, which may be particularly problematic for people living in rental accommodation. In such cases, these issues may increase risk of relinquishment [7]” (see new lines 56-59) It may be helpful to discuss why there is poor owner adherence to programmes to reduce SRB (Line 66-67).	2	1	My main question when reading this section concerned how owners would know that certain separation-related behaviors occurred when they were away?			ani12040482_perova	0
+The following text has now been added beginning at line 69: “Whilst the precise reasons for poor adherence to advice for preventing or reducing SRBs have not been documented, it is at least partly attributed to the relative difficulty of adhering to specific aspects of these programmes, with the easier pieces of advice followed more often than the parts that take more commitment and consideration [13].” I wasn't sure if line 79-80 was needed- about dogs having returned to pre-pandemic pattern, I think it is a little dismissive of your own research which is valuable and applicable beyond the pandemic.	2	1	We have addressed your comments and responded in the attached file.			ani12040482_perova	0
+Thank you for your comment, I think we were pre-judging that people would think it was just another covid paper, and we are glad to see you agree with us that it is valuable beyond the pandemic. This line has now been deleted.	2	1	I felt the discussion about changes in greeting behaviour as an early indicator was interesting and could be expanded.			ani12040482_perova	0
+The findings were all extremely similar, so it felt quite repetitive to discuss them all. We actually had more detail in an early draft then trimmed it out. However, we have now added some more of this detail as requested (see lines 100 to 112), whilst trying to limit repetition.	2	1	The authors gathered data (via owner surveys) about the same dogs before the pandemic, at the beginning of the pandemic and approx.			ani12040482_perova	0
+We presumed that at some point owners would pop out of the house, to take out the bin or go to the garage etc.. Such leaving events are likely to be experienced differently by some dogs, as the owners won’t likely give off the same cues as when they leave for an extended period of time, such as picking up house keys, or leaving through a different door. This has been explained now in lines 179-83.	2	1	This Figure should be moved to the Supplement material as it tells nothing about the dogs’ behaviour.			ani12040482_perova	0
+Thank you, this seemed ok when we submitted it, but must have gone wrong when uploaded but we’ve corrected it now.	2	1	Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+We’ve amended description of the study periods in a few places, including between lines 132-142, which we hope addresses this query now.	2	1	Here are some suggestions that can be made throughout the paper:			ani12040482_perova	0
+One of the other reviewers also commented on this and asked for Figure 1 and much of the text to be moved to the supplemental material, so It’s also further explained in the supplementary material how it links to working and in lines 140-142 in the main text which now explains that many businesses were open again in October.	2	1	The article, “Impact of Changes in Owner Leaving Pattern on Separation-Related Behaviour in Pet Dogs in the UK” investigated whether the prevalence of SRB in dogs changed with changes in dog management during and after COVID lockdowns.			ani12040482_perova	0
+This was due to the errors you mentioned below. We used the Word automatic referencing and somehow that seemed to get broken when the document was uploaded to the online system. It’s been done manually now.	2	1	The authors still take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed.			ani12040482_perova	0
+See above.	2	1	The entire first paragraph of Discussion repeats what has already been written in earlier chapters (line 372-379).			ani12040482_perova	0
+Agreed, this was how we interpreted why the owner was reporting this change. We have rephrased this section now.	2	1	According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			ani12040482_perova	0
+Thank you for your review. We have expanded the discussion to address these points, however due to another reviewer saying the paper was already too long, we have kept this brief. A section detailing relevance to practitioners has been added to lines 545-551.	2	1	Specific points requiring attention: L18-20 „Whether dogs showed SRBs or not changed considerably over the months of the study, and one in ten dogs were found to have developed new SRBs in October, that they didn’t show before the pandemic” Overall, the percentage of dogs showing SRBs decreased during the study, and this should be clarified in the sentence above.			ani12040482_perova	0
+Yes, this is correct. This is stated in line 232-233 or the original submission, but it seems we omitted it from the Methods section. Thank you for spotting that. Text has been added to lines 198-200 to explain this.	2	1	L118 Materials and Methods This section (after the Ethics) should begin with the Subjects section.			ani12040482_perova	0
+Within the results table (Table 2) the direction of the estimate for this result is negative. Whilst we agree that it would be more impactful worded the other way around, we worry that changing the way it is described won’t match up to the data as it was coded and reported. We haven’t changed this wording for these reasons.	2	1	We have addressed your comments and responded in the attached file.			ani12040482_perova	0
+Yes, and no, depending on your perspective. Whilst this was a significant predictor in the modelling, in reality 55% of dogs that were being left again changed from SRB+ to SRB-. So whilst 45% of dogs still showed it, a large proportion did not.	2	1	Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing.			ani12040482_perova	0
+Thank you for these thoughtful comments, we agree that we were a bit overly focussed on the results that supported the studies hypothesis and neglected to discuss the findings related to the dogs whose SRB appeared to have resolved. The number and percentage are detailed in the results section (lines 282-283 of the original submission and 298-298 in this one), but we failed to discuss these in the Discussion. We ran two models, one focussing on the dogs who started SRB- to see how many changed to SRB+ or remained SRB- and what factors influenced this, and the other focussing on dogs that started SRB+ to see what factors influence whether they changed to SRB- or remained SRB+. These models therefore, cover the comparisons you suggest (unless we’ve misinterpreted your comments). We have now added some discussion of the dogs who began the pandemic SRB+ however, as this was lacking from the first version (see the paragraph beginning at line 524).	2	1	noise complaints and issues with housing; property damage; decline of human-pet bond) could be beneficial (line 56-57).			ani12040482_perova	0
+Thank you for this comment, we have discussed this further in the Discussion now (see lines 517-523)	2	1	Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+Thank you, we have changed the title to: Impact of changes in time left alone on separation-related behaviour in UK pet dogs	2	1	L232-242 These sections should be moved to the Materials and Methods as a “Subjects” section			ani12040482_perova	0
+This is how it was phrased in the survey, so this has not been changed.	2	1	Materials and Methods: Line 151: how was the criterion of “at least 5 minutes” chosen?			ani12040482_perova	0
+Thank you, this edit has been applied.	2	1	Larger issues: I would like to see the authors address a few different angles in the discussion 1.			ani12040482_perova	0
+It seemed unfeasible to expect a dog to never be left for even a minute or two, for example if the owner pops outside to take out the bin or to get something from the garage/shed etc.	2	1	This is discussed later on but highlighting here would be useful.			ani12040482_perova	0
+The wording given to the owners is all presented here, and in the supplementary material. No further definitions for behaviours or vocalisations were given. Pined, whined and cried were all mentioned, as all are words owner tend to use to describe similar dog vocalisations. As with all owner awareness of SRBs, it is fundamentally challenging unless they use recording equipment to view their dogs when they leave them or are otherwise able to view their dogs when alone such as by looking through a window. As this is a limitation of the method used, the appropriate place to mention this would be in the Discussion, not the Methods, so we have not discussed this limitation there.	2	1	This is a great figure, thank you for including it.			ani12040482_perova	0
+The wording of the question asked about the dog being left alone, without human company (see line 173), so it applied to anyone, and specifically meant when no-human was present in the house with the dog.	2	1	There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing?			ani12040482_perova	0
+This has now been explained, however this section has been moved entirely to the supplementary material on request of other reviewers.	2	1	We might see some dogs become sensitized over time and develop SRB (or something else changes in their life to precipitate it), while others might habituate or their behavior undergo extinction.			ani12040482_perova	0
+Not strictly speaking, no. Dogs who were not left in February were excluded from the analysis. So, whilst there are 0 of them, that’s because of methodological choice not because there weren’t any dogs left alone. A note explaining this has been added to the figure legend.	2	1	Line 96-97- some more detail on the findings of these various surveys would be beneficial similarly to Bowen's study, just so provide a clear background/discussion of literature setting scene for your study.			ani12040482_perova	0
+There were issues in the figure referencing when we uploaded the manuscript to the submission system. This has now been corrected.	2	1	Larger issues: I would like to see the authors address a few different angles in the discussion 1.			ani12040482_perova	0
+For some reason I was convinced I’d read it was optional for longer discussions, but having double checked, you are indeed correct. Thank you. A brief conclusion section has now been added.	2	1	Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+Thank you, we have corrected the typo.	2	1	L314 Table 2: bold is not used here although it was used in Table 1			ani12040482_perova	0
+This has been corrected	2	1	Manuscript has been slightly improved, but my comments were not included.			ani12040482_perova	0
+Thank you for your comments. The detail was provided to anchor the study in the background of what was occurring at the time, but we agree it is not necessary to the reader to see in the main text. These details and Figure 1 have been moved to the Supplementary material.	2	1	Materials and Methods: Line 151: how was the criterion of “at least 5 minutes” chosen?			ani12040482_perova	0
+We agree this was completely missed from the Discussion. It wasn’t intentional, we were just overly focussed on the results that related to the research hypothesis. Discussion around this has been added now to lines 524-544.	2	1	This Figure should be moved to the Supplement material as it tells nothing about the dogs’ behaviour.			ani12040482_perova	0
+Agreed, we have altered this part of the simple summary and abstract and included discussion of this in lines 539-544 now as well as into the conclusions, thank you.	2	1	Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+Leaving time has been changed throughout to ‘time left alone’ L33 10th instead of “10th” Thank you, we’ve corrected the typo.	2	1	There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing?			ani12040482_perova	0
+Thank you for pointing this out. We define SRB in the opening lines of the Introduction (45-50), however you are right that in places the terms can get confusing. Section 3.7.3. details behaviours owners reported as associated with separation, that are not what class as SRBs. Those were names ‘other’ separation behaviour but this has now been clarified, hopefully, by remaining the section ‘Other’ separation-linked behaviour, and then in the text it is now stated they are not SRBs. The term ‘separation anxiety’ was used in the introduction where it referenced a study that used that specific term. This has now been put into quote marks to be clear it is their term, not ours. The one mention of separation-distress has also now been changed.	2	1	Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing.			ani12040482_perova	0
+Thank you, we’ve corrected the typo.	2	1	The article is generally well-written, the topic is timely, important, and interesting from a dog welfare aspect.			ani12040482_perova	0
+This has now been discussed further in lines 98-100.	2	1	This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			ani12040482_perova	0
+This has now been moved to the beginning of the methods section.	2	1	This is good news, and it is unclear for me why is it reported as bad news.			ani12040482_perova	0
+Agreed, this has been amended and condensed, thank you.	2	1	L291 The odds ratio (OR) is 5.38 and not 4.38 in Table 1.			ani12040482_perova	0
+Thank you, this seemed ok until the manuscript was uploaded but we’ve corrected it now.	2	1	Introduction: This is clear and in general provides a good background to the topic (bar a couple of points where more expansion would be beneficial).			ani12040482_perova	0
+We asked about the dogs’ management and behaviour over the previous 7 days so as to limit recall bias and anchor the data to a specific snapshot of time, rather than being more general. This was first asked for the first survey which was specifically looking at changes in management and behaviour during the strict lockdown period, so needed to be time specific. Questions were then repeated in the follow-up survey. Wording has been added to lines 165-169 now to clarify this.	2	1	The manuscript: Impact of Changes in Owner Leaving Pattern on Separation- Related Behavior in Pet Dogs in the UK is one more paper on the behavior of dogs in the COVID-19 pandemic developed in the last two years.			ani12040482_perova	0
+this has been done now.	2	1	This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			ani12040482_perova	0
+Thank you for spotting this, we have updated both the methods (L243-247) and results section (L370-374) with further detail of inter-rater reliability checks.	2	1	As I wrote in the previous manuscript review:			ani12040482_perova	0
+At the end of survey 1 there was the option for people to provide their demographic details, which not everyone did. This was done to conform to GDPR by asking only the most necessary questions. Given that the research question here is related to the dogs’ behaviour change over time versus leaving hours, it wasn’t deemed relevant to include analysis of the owner demographics.	2	1	Conclusions: I believe Animals requires a Conclusions section.			ani12040482_perova	0
+This has now been done.	2	1	L314 Table 2: bold is not used here although it was used in Table 1			ani12040482_perova	0
+For the purpose of this study it was required that we had baseline data on the dogs’ leaving behaviour so that we could classify them as SRB+ or – before the pandemic began. Owners of dogs who were not left alone in a typical week in February, were not asked about their dogs’ separation-related behaviour, so we did not have these data for them which is why they were excluded. New text has been added to the Subjects section of the methodology to explain how many owners answered the follow up questionnaire, the response rate and what percentage and number of dogs were not left home alone before the pandemic. The new section beginning at line 146 now reads: “A total of 2,425 people completed the follow-up survey for the same dog they scored in the original survey (a 51.9% response rate), of which 2,285 had previously provided data to indicate whether their dog was left alone or not in February 2020. Of these, 15.3% dogs (n=305) were not left alone at all in an average week in February, and as such, these dogs did not have baseline data for their behaviour when left so were excluded from further analysis.” Additionally, the number of people we emailed the invitation to (n=4,670) has been added to line 139.	2	1	“More Attention than Usual”: A Thematic Analysis of Dog Ownership Experiences in the 530 UK during the First COVID-19 Lockdown. Anim. 2021, Vol. 11, Page 240 2021, 11, 240, doi:10.3390/ANI11010240).			ani12040482_perova	0
+For the 1407 dogs who were clear of SRB in February, they changed from being 100% clear to being 90.1% clear, with 1 in 10 of these dogs developing SRB they didn’t have previously. However, because of the dramatic improvement seen in the dogs that had SRB before the pandemic, the overall prevalence of SRB in the population decreased from 22% in February, to 17.2% in October (reported in the paragraph above). The numbers have been clarified in this section to help future readers, and additional discussion has been added around the overall decrease in SRB prevalence in both the abstract and in the discussion (lines 524-544).	2	1	The next section (l281-283) emphasizes that 55.7% of the dogs who were reported to show SRB in February were clear in October.			ani12040482_perova	0
+Thank you, we’re glad you liked it! Further explanation to help interpret the figure has now been added.	2	1	That dogs showing SRB prior to lockdown were more likely to show it after.			ani12040482_perova	0
+Thank you for spotting that!	2	1	However, your results that the greater the change → the greater the new risk of SRB does lend support to the idea that this change in management might be a factor.			ani12040482_perova	0
+This was explained further down in the section on the multi-variable model, but I have now moved the explanation to here, where it’s first mentioned and added some additional text also.	2	1	Thank you for your comments and for the constructive feedback.			ani12040482_perova	0
+Thank you, this has been added now.	2	1	There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing?			ani12040482_perova	0
+Thank you! Looks like I read the z-value when I was typing. I promise this has been proof read, but it’s so easy to miss things, thank you so much!	2	1	L232-242 These sections should be moved to the Materials and Methods as a “Subjects” section			ani12040482_perova	0
+Since there were no non-significant variables it wasn’t necessary to distinguish the results.	2	1	(*Christley, R.M.; Murray, J.K.; Anderson, K.L.; Buckland, E.L.; Casey, R.A.; Harvey, N.D.; Harris, L.; Holland, K.E.; McMillan, 523 K.M.; Mead, R.; et al.			ani12040482_perova	0
+This has been corrected, thank you.	2	1	Line 214: if possible (might not be possible if this was how it was phrased in the survey), change “different to before” to “different from before”			ani12040482_perova	0
+Thank you for this, we were familiar with the paper but had missed that link. New text has been added here now to discuss this.	2	1	Specific points requiring attention: L18-20 „Whether dogs showed SRBs or not changed considerably over the months of the study, and one in ten dogs were found to have developed new SRBs in October, that they didn’t show before the pandemic” Overall, the percentage of dogs showing SRBs decreased during the study, and this should be clarified in the sentence above.			ani12040482_perova	0
+Agreed, this has been explicitly stated here now.	2	1	The entire first paragraph of Discussion repeats what has already been written in earlier chapters (line 372-379).			ani12040482_perova	0
+Thank you so much for your time and these comments, they have really helped to strengthen the paper and we really appreciate it.	2	1	That dogs showing SRB prior to lockdown were more likely to show it after.			ani12040482_perova	0
+In order to address your feedback fully, we felt it necessary to split Figure 7. The data contained in the original Figure 7 is now split between Figure 7 (deals with CD14 and CD68 only) and Figure 8 (deals with CD11b and CD36).	2	1	The red population shows positive binding of anti-CD36 and anti-CD11b to the U937 cells (gated on the previous plot S4A, FSC/SSC).			antiox11040683_makarova	0
+Fig 7b. We have removed the Giemsa images and replaced with phase contrast images to better show cell morphology under our light microscope. We also include representative flow cytometric overlayed histograms, which display CD14 and CD68 expression changes between our Vector control and WT / CXXS BCAT1 cells. Included on these histograms is the mean fold MFI change (ratio).	2	1	The Authors showed the role of the BCAT1 CXXC motif in AML and to understand how this may influence cellular redox processes.			antiox11040683_makarova	0
+Fig S4B. This is fully compensated.	2	1	These isotype controls were used to set the negative thresholds, as indicated by the quadrants on the plot.			antiox11040683_makarova	0
+Fig 4d. This is going in the correct direction.	2	1	This additional narrative is added at line 645-660 and highlighted in yellow on the resubmission.			antiox11040683_makarova	0
+Thank you for taking time to review are work and for giving us the opportunity to comment on the use of U937 cells as a limitation for the work presented. We have included a section dedicated to addressing this in the discussion section, along with supporting references.	2	1	The red population shows positive binding of anti-CD36 and anti-CD11b to the U937 cells (gated on the previous plot S4A, FSC/SSC).			antiox11040683_makarova	0
+"We agree that transient analysis is important in designing a robot joint. However, the transient analysis requires relevant parameters of the robot arm be determined besides the parameters of the joint. The following sentence has been added at the end of Conclusions: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors"""	2	1	[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			app12052642_perova	0
+The mentioned reference is focused on an analytical method for modeling large deflections in compliant mechanisms, while this work presents a method of enhancing the tensile strength of lamina emergent joints by using double-layer materials. As compared to Ref. [19] which focuses on concepts and demonstrations, this work mainly focuses on analytical modeling of one design concept and provides finite element and experimental validations. The following sentences have been added	2	1	Author Response We thanks the reviewer for the encouragement and valuable comments.			app12052642_perova	0
+"As to the second comment, Ref. [34] has been new added in the reference list and the following sentence has been added at the end of the second paragraph of Introduction: ""Ref. [34] made a robot using double-layer LEMs, which is driven by magnetic force. This paper further discussed how to adjust the motion by changing different parameters of the mechanism"". The following sentence has been added at the end of Conclusion: ""For future work we will develop variable stiffness techniques of this joint, and explore its application in a robot arm considering both kinetostatic and transient behaviors."""	2	1	The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			app12052642_perova	0
+We have rewritten the conclusions and abstract, and added additional keywords.	2	1	Author Response We thanks the reviewer for the encouragement and valuable comments.			app12052642_perova	0
+"at the beginning of the last paragraph of Introduction: ""In this work, based on the design concept of improving the tensile stiffness of LET joints in Ref. [19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint. A theoretical model is derived for the DL-LET joint and verified by FEA and experimental results."" The following sentences have been added in the first paragraph of Introduction to mention the base paper and the related work (more references have been added): ""Finding suitable compliant joints [7] can be a key step in designing LEMs. Jacobsen et al. [8] first proposed the design concept of utilizing torsional deflection to obtain out-of-plane motions from planar structures, leading to a new group of compliant joints for LEMs called Lamina Emergent Torsional (LET) joints""."	2	1	Nice piece of research but one aspect needs to be addressed.			app12052642_perova	0
+The references have been reformatted according to standard journal format.	2	1	Following the reported physical experiments (Fig.			app12052642_perova	0
+We have added more relevant references in this version.	2	1	Author Response Thanks for your encouragement and suggestion.			app12052642_perova	0
+The contribution of this work with respect to the relevant work is further described in Introduction (as marked in the manuscript).	2	1	The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			app12052642_perova	0
+The theoretical part has been extended in the modeling section (page 3).	2	1	Author Response We thanks the reviewer for the encouragement and valuable comments.			app12052642_perova	0
+The modeling section (page 3) has been extended to make it clear how the parameters were obtained.	2	1	Author Response Thanks for your encouragement and suggestion.			app12052642_perova	0
+The recommended and other relevant references have been added.	2	1	Chen, G.; Ma, F.; Hao, G.; Zhu, W. Modeling large deflections of initially curved beams in compliant mechanisms using 277 chained Beam-Constraint-Model, ASME J. Mech.			app12052642_perova	0
+Subsection 3.2 has been rewritten to add more details regarding the obtained results.	2	1	Robot., 2019, 11: 011002, https://doi.org/10.1115/1.4041585.			app12052642_perova	0
+The following sentence has been added at the end of first paragraph of Conclusions to clarify disadvantage of DL-LET: “The DL-LET joint is vulnerable to delamination of the two layers.”	2	1	The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			app12052642_perova	0
+Thank you for your explanation.	3	2	Nice piece of research but one aspect needs to be addressed.			app12052642_perova	0
+The authors answered to all my concern in the second version of the paper. In conclusion the paper deserves to be acceped as contribution in Applied Science journal.	3	2	[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			app12052642_perova	0
+We have added all the new references suggested by Referee 2 on page 3 of the new manuscript. These includethe ones mentioned in our reply to Referee 1’s point 4 above, and all also the references: Corney98,Mekhov09,leroux10,Mekhov12,Niedenzu13,Lee14.	2	1	(b) There will be heating limiting the available measurement time.			atoms4010002_makarova	0
+Concerning the referee’s other point, about the suitability of this system to measure weak forces, we first note that the use of Bloch oscillations without a cavity is proving to be an excellent way to measure gravity (e.g. the experiments by G.M. Tino’s group).	2	1	In my opinion, the main interest of the work is in the cold atoms and cavity dynamics, continuous observation, and in the specific optomechanical dynamics.			atoms4010002_makarova	0
+Concerning the question of whether we have taken the effects of higher Bloch bands into effect, our numerical calculations do take this into account. The question of higher Bloch bands is a little subtle when it comes to our analytical calculation done in the Wannier-Stark picture. In the presence of the force, the true eigenstates of the system are no longer the Bloch states but the localized Wannier-Stark states. In the analytical calculation we restrict ourselves to just the ground band Wannier-Stark states. However, even with this approximation we find very good agreement between the numerical and analytical calculations.	2	1	Author Response The authors present a new interpretation to study Bloch oscillations in a cavity sustained optical lattice and the potential use of this system for very sensitive force measurement.The idea is closely related to previous work as Ref.			atoms4010002_makarova	0
+Done. We have re-written the first part of the abstract as: “In this paper we give a new description, in terms of optomechanics, of previous work on the problem of an atomic Bose-Einstein condensate interacting with the optical lattice inside a laser-pumped optical cavity and subject to a bias force such as gravity \cite{Ven09,Ped09,Gol14}. An atomic wave packet in a tilted lattice undergoes Bloch oscillations; in a highfinesse optical cavity the backaction of the atoms on the light leads to a time-dependent modulation of the intracavity lattice depth at the Bloch frequency which can in turn transport the atoms up or down the lattice.”	2	1	Following the referee’s suggestion we have added the reference to Mekhov’s paper.			atoms4010002_makarova	0
+We have added the following sentences on p3 (and four new references, including the one to Gangl’s work, in the bibliography): “A number of experiments have already demonstrated how the light transmitted by a cavity can be used to track the motion of atoms trapped inside \cite{Hood98,Hood00,Pinkse00}, and in particular, a theoretical analysis of the information stored in the frequency spectrum has been given in \cite{Gangl00}, showing that atomic motion introduces sidebands either side of the pump frequency. In our case, Bloch oscillations at angular frequency $\omega_{B}$ generates sidebands separated from the pump frequency by $\pm \omega_{B}$ (and harmonics thereof in the strong coupling regime).Because the Bloch frequency is proportional to the applied force $\omega_{B}= F d/\hbar$, where $d=\lambda/2$ is thelattice period, a detection of the spectrum of the transmitted light gives $F$ directly.”	2	1	(2) It is very helpful to connect cavity output spectra and forces.			atoms4010002_makarova	0
+Actually, we do not claim that oscillation frequencies in general will be unaffected by the backaction. Rather, we claim that one particular frequency, the Bloch frequency, will be unaffected.	2	1	The idea is closely related to previous work as Ref.			atoms4010002_makarova	0
+Concerning the change in wavelength (and hence change in lattice constant) due to the refractive index of the gas, this effect is tiny. We consider it in section 6, on p17 and show that the correction to the refractive index is of order 10^-9 for our parameters. This is to be distinguished from the effective change in the cavity length which, due to the resonance effect, is substantial and is precisely what lies behind the amplitude modulation of the lattice.	2	1	The estimated accuracy is $10^6$ that does not sound great.			atoms4010002_makarova	0
+Yes we agree that there will be extra noise due to the open nature of the cavity, and we already refer to this on p3 of the Introduction: “The disadvantage of working in a cavity is that quantum measurement backaction, in the form of random fluctuations in the cavity field due to photons spontaneously leaking out of the cavity, heats up the cold atoms and limits the coherence time of the measurement \cite{Ven13}.” where the cited paper is one of ours [Prasanna Venkatesh, B.; O’Dell, D. H. J.; Bloch oscillations of cold atoms in a cavity: Effects of quantum noise. Phys. Rev. A 2013, 88, 013848] where we directlycalculated the heating effect for Bloch oscillating atoms in a cavity within the Bogoliubov approximation.	2	1	Some rough estimate on the time scales should be added.			atoms4010002_makarova	0
+The coherence time for the many-particle Bloch oscillating state is in fact very hard to calculate (mainly because even the coherent part is time-dependent rather than stationary). It can, however, be estimated and so we have added in the following sentences on p3:“\textcolor{red}{The coherence time is particularly hard to calculate in the Bloch oscillating case \cite{Ven13} due to the time dependence introduced by the Bloch oscillations, especially in the presence of many particles, but it can be roughly estimated to be $\tau=\tau_{\mathrm{sp}}/(1+C)$ \cite{Ven09} at cavity resonance, where$\tau_{\mathrm{sp}}^{-1}=2 \gamma \vert \alpha \vert^2 \Omega_{0}^2/\Delta_{a}^2$ is the spontaneous emission rate at an antinode. The factors $\vert \alpha \vert^2$ and$\Delta_{a}$ are the mean number of cavity photons and the detuning of the laser from atomic resonance, respectively, and will be properly defined in the next section. The numerical value of $\tau$ for the parameters considered in this paper will be given in Section \ref{sec:metrology}. Of course, Bose-Einstein condensates can be continuously measured and used for sensingwithout a cavity, e.g.\ \cite{Ruostekoski98,Dalvit02,Saba05,Lee12,Java13}, but the cavity case is particularly interesting because it allows for a strong atom-light interaction even in the quantum regime.} “In addition, in Section 6 (paragraph 4, Pg 18) we have provided further details required to estimate the coherence time for the parameters used in the paper. We find thatcoherence times of up to a couple of seconds (many thousands of Bloch oscillation periods) are possible	2	1	In particular for the blue detuned case not only the average momentum but also the momentum spread will grow.			atoms4010002_makarova	0
+We have added all the new references suggested by Referee 2 on page 3 of the new manuscript. These includethe ones mentioned in our reply to Referee 1’s point 4 above, and all also the references: Corney98,Mekhov09,leroux10,Mekhov12,Niedenzu13,Lee14.	2	1	Nevertheless the presentaion sheds new light onto the underlying microscopic dynamics and connects the dynamics to proven optomechanical models of cavity BEC dynamics.			atoms4010002_perova	0
+Concerning the referee’s other point, about the suitability of this system to measure weak forces, we first note that the use of Bloch oscillations without a cavity is proving to be an excellent way to measure gravity (e.g. the experiments by G.M. Tino’s group). The sensitivity of their measurements, after many years of development, are only one order of magnitude above our estimated sensitivity, and their measurements take one hour whereas ours should take one second which means it might have applications in rather different situations (e.g. local gravity mapping while flying). Our work is very much still in progress and this paper will not, we hope, be our final word on this subject. Indeed, we point out a possible way forward in the last section that builds on the insight gained from the optomechanics analogy. It seems likely that the homodyne phase measurement regime, where backaction can be reduced, will likely be more favourable when it comes to absolute sensitivity than photon counting type measurement we focus on at the moment. For these reasons we would like to keep the measurement aspects of the paper intact: we would like to advertise them and, perhaps, be a little provocative.	2	1	Inparticular for the blue detuned case not only the average momentum but also the momentum spread will grow.			atoms4010002_perova	0
+Concerning the question of whether we have taken the effects of higher Bloch bands into effect, our numerical calculations do take this into account. The question of higher Bloch bands is a little subtle when it comes to our analytical calculation done in the Wannier-Stark picture. In the presence of the force, the true eigenstates of the system are no longer the Bloch states but the localized Wannier-Stark states. In the analytical calculation we restrict ourselves to just the ground band Wannier-Stark states. However, even with this approximation we find very good agreement between the numerical and analytical calculations.	2	1	Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.			atoms4010002_perova	0
+Concerning the question of radiationpressure forces, we take them into account on the atoms but not on the mirrors.	2	1	(3) The authors claim there should be no backaction of the field on the oscillation frequencies.			atoms4010002_perova	0
+Done. We have re-written the first part of the abstract as: “In this paper we give a new description, in terms of optomechanics, of previous work on the problem of an atomic Bose-Einstein condensate interacting with the optical lattice inside a laser-pumped optical cavity and subject to a bias force such as gravity \cite{Ven09,Ped09,Gol14}. An atomic wave packet in a tilted lattice undergoes Bloch oscillations; in a highfinesse optical cavity the backaction of the atoms on the light leads to a time-dependent modulation of the intracavity lattice depth at the Bloch frequency which can in turn transport the atoms up or down the lattice.”	2	1	(2) It is very helpful to connect cavity output spectra and forces.			atoms4010002_perova	0
+We have added the following sentences on p3 (and four new references, including the one to Gangl’s work, in the bibliography): “A number of experiments have already demonstrated how the light transmitted by a cavity can be used to track the motion of atoms trapped inside \cite{Hood98,Hood00,Pinkse00}, and in particular, a theoretical analysis of the information stored in the frequency spectrum has been given in \cite{Gangl00}, showing that atomic motion introduces sidebands either side of the pump frequency. In our case, Bloch oscillations at angular frequency $\omega_{B}$ generates sidebands separated from the pump frequency by $\pm \omega_{B}$ (and harmonics thereof in the strong coupling regime).Because the Bloch frequency is proportional to the applied force $\omega_{B}= F d/\hbar$, where $d=\lambda/2$ is thelattice period, a detection of the spectrum of the transmitted light gives $F$ directly.”	2	1	(3) The authors claim there should be no backaction of the field on the oscillation frequencies.			atoms4010002_perova	0
+Actually, we do not claim that oscillation frequencies in general will be unaffected by the backaction. Rather, we claim that one particular frequency, the Bloch frequency, will be unaffected. The Horak paper shows that Bloch energies (i.e. eigenmodes for a particle in a periodic potential…not to be confused with the Bloch oscillation frequency) will be changed by the backaction. This is because the presence of atoms in the cavity shifts the cavity resonance and hence the depth of the intracavity lattice: the Bloch energies are dependent on the potential depth. This change in potential depth due to backaction is fully included in our treatment. However, a change in the depth of a periodic potential does not change the Bloch frequency because this depends on the lattice’s spatial period, not the depth. Because this key aspect of our treatment can clearly lead to confusion we have modified the following sentence in the conclusions on p19, and added the reference to Horak’s paper: “Chief among these is that the backaction does not alter the frequency of the Bloch oscillations. By contrast, in the harmonic oscillator case there is the so-called optical spring effect which gives a dependence of oscillator frequency on field amplitude and detuning. To be clear, other motional frequencies are altered: because the intracavity lattice depth is modulated by the backaction this will affect certain types of atomic motion, for example the oscillation frequency of an atom about the bottom of one of the potential minima \cite{Horak00}. Nevertheless, the Bloch oscillation frequency is robust against this depth modulation because it only depends on the lattice period, not its depth.” Concerning the change in wavelength (and hence change in lattice constant) due to the refractive index of the gas, this effect is tiny. We consider it in section 6, on p17 and show that the correction to the refractive index is of order 10^-9 for our parameters. This is to be distinguished from the effective change in the cavity length which, due to the resonance effect, is substantial and is precisely what lies behind the amplitude modulation of the lattice.	2	1	B: Atomic, Molecular and Optical Physics, 45(10), 102001).			atoms4010002_perova	0
+Yes we agree that there will be extra noise due to the open nature of the cavity, and we already refer to this on p3 of the Introduction: “The disadvantage of working in a cavity is that quantum measurement backaction, in the form of random fluctuations in the cavity field due to photons spontaneously leaking out of the cavity, heats up the cold atoms and limits the coherence time of the measurement \cite{Ven13}.” where the cited paper is one of ours [Prasanna Venkatesh, B.; O’Dell, D. H. J.; Bloch oscillations of cold atoms in a cavity: Effects of quantum noise. Phys. Rev. A 2013, 88, 013848] where we directlycalculated the heating effect for Bloch oscillating atoms in a cavity within the Bogoliubov approximation. Following the referee’s suggestion we have added the reference to Mekhov’s paper.The coherence time for the many-particle Bloch oscillating state is in fact very hard to calculate (mainly because even the coherent part is time-dependent rather than stationary). It can, however, be estimated and so we have added in the following sentences on p3:“\textcolor{red}{The coherence time is particularly hard to calculate in the Bloch oscillating case \cite{Ven13} due to the time dependence introduced by the Bloch oscillations, especially in the presence of many particles, but it can be roughly estimated to be $\tau=\tau_{\mathrm{sp}}/(1+C)$ \cite{Ven09} at cavity resonance, where$\tau_{\mathrm{sp}}^{-1}=2 \gamma \vert \alpha \vert^2 \Omega_{0}^2/\Delta_{a}^2$ is the spontaneous emission rate at an antinode. The factors $\vert \alpha \vert^2$ and$\Delta_{a}$ are the mean number of cavity photons and the detuning of the laser from atomic resonance, respectively, and will be properly defined in the next section. The numerical value of $\tau$ for the parameters considered in this paper will be given in Section \ref{sec:metrology}. Of course, Bose-Einstein condensates can be continuously measured and used for sensingwithout a cavity, e.g.\ \cite{Ruostekoski98,Dalvit02,Saba05,Lee12,Java13}, but the cavity case is particularly interesting because it allows for a strong atom-light interaction even in the quantum regime.} “In addition, in Section 6 (paragraph 4, Pg 18) we have provided further details required to estimate the coherence time for the parameters used in the paper. We find thatcoherence times of up to a couple of seconds (many thousands of Bloch oscillation periods) are possible	2	1	Nevertheless the presentaion sheds new light onto the underlying microscopic dynamics and connects the dynamics to proven optomechanical models of cavity BEC dynamics.			atoms4010002_perova	0
+The review article by W Nakel and C. T. Whelan you refer to do indeed tackle the relativistic regime but at the expense of a huge amount of numerical work.	2	1	1) Earlier publications [Physics Reports 315 (1999) 409 and related references], a fully relativistic distorted-wave Born approximation (rDWBA) has been developed to describe the relativistic ionization processes.			atoms4010010_makarova	0
+The work of W Nakel and C. T. Whelan is devoted to study heavy atoms but Our work is aimed to study analytically the hydrogen atom, we believe that such a comparison is impossible.	2	1	The work and discussion are very short, but still interesting.I recommend it for publication after the authors answer the questions and make the corrections described below:			atoms4010010_makarova	0
+We are not in a position to compare with these works since they are devoted to heavy atoms.	2	1	(1) and (2) for Vd: in the second term of the right hand side (rhs) the quantities between the vertical bars are vectors and the notation should be different from that of the first term of the rhs, which represent a modulus.			atoms4010010_makarova	0
+Points (i) and (ii) have been corrected and changed following to the referee’s amendments.	2	1	My overall recommendation is that after the listed items are addressed the manuscript be accepted for publication without further review.			atoms4010010_makarova	0
+A major revision of the paper has been carried out to take all the comments, suggestions and english corrections into account.	2	1	------------------------------------------- Answer of authors ------------------------------------------- The changes is made in the text.			atoms4010010_makarova	0
+The changes is made in the text.	2	1	The results confirm predictions of other theories.			atoms4010010_makarova	0
+The review article by W Nakel and C. T. Whelan you refer to do indeed tackle the relativistic regime but at the expense of a huge amount of numerical work. So, we think that presenting for the first time a work that incorporates fully analytical results and QED formalism is interesting to the scientific community. The work of W Nakel and C. T. Whelan is devoted to study heavy atoms but Our work is aimed to study analytically the hydrogen atom, we believe that such a comparison is impossible. Another difference that makes comparison difficult is that the work of W Nakel and C. T. Whelan uses coplanar asymmetric geometry while in our work we use coplanar symmetric geometry.	2	1	· Line 108: don’t capitalize “triple” · Line 112 and caption for Table I: insert “the” before “TDCS: and “peaks” and change “peak’s” to “peak” without italics · Line 115: delete “for positron impact” at end of sentence.			atoms4010010_perova	0
+We are fully aware that the mainstream trend is to study heavy atoms. But some fundamental processes have not until now been investigated fully and analytically and it is our very humble belief that such a fundamental work must be presented to the international scientific community. We are not in a position to compare with these works since they are devoted to heavy atoms.	2	1	5 an 6 are mentioned but only 5 is discussed.			atoms4010010_perova	0
+Points (i) and (ii) have been corrected and changed following to the referee’s amendments.	2	1	I find no major problems with the manuscript, either in content or presentation.			atoms4010010_perova	0
+As you requested, we have made all necessary changes in our manuscript. A major revision of the paper has been carried out to take all the comments, suggestions and english corrections into account. We believe that the paper has been significantly improved.	2	1	I suggest combining the figures to parts a and b and modifying the text as needed.			atoms4010010_perova	0
+The changes is made in the text.	2	1	------------------------------- 1) Answer of authors -------------------------------			atoms4010010_perova	0
+We added references to the Wolf and Tourrencin and A. Peters, K.Y. Chung and S. Chu articles. The latter of these articles does not address the quantum corrections 2.	2	1	Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			atoms4020014_perova	0
+The referee implies that the leading term is not a quantum correction owing to the cancellation of a numerator proportional to hbar and a denominator in the spatial phase factor that is also proportional to hbar. This is, is some respect, a question of semantics, and has a long history in discussions of quantum effects in atom interferometry. Basically this term is a Doppler shift, which is clearly of classical origin, with no need to impose arguments related to cancellations of hbars in numerators and denominators. In the approach we follow using the Wigner representation, there are no numerators and denominators; the leading term has no hbar.	2	1	The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			atoms4020014_perova	0
+The density matrix in the Wigner representation has been widely used in problems related to the recoil effect and laser cooling. For some mysterious reason it hasn’t used in much in problems involving atom interferometry. Of course there are hundreds of articles (maybe more) and books on the Wigner representation; however here we are concerned mainly with its application to calculations involving atom interferometry. The only examples we know of are the ones referred to in the paper.	2	1	Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			atoms4020014_perova	0
+The article by Fam is devoted to the use of Wigner representation for the two-particle density matrix, which is outside the scope of our article. Moreover, in contrast to that article, we do not use an expansion of the equation for the Wigner distribution in powers of hbar. For both these reasons, we do not include a reference on this article 5.	2	1	The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			atoms4020014_perova	0
+As far as we can tell, the phi_Q term is not related directly to the spreading of the wave-packet. It arises as a quantum correction to phase factors during periods of free evolution. At this point, we do not have any other physical interpretation. We do not know yet how this term would manifest itself in the other approaches (path-integrals, ABCD theorem and etc.). This remains an interesting topic for future work, but is outside the scope of our article.	2	1	The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+We added a paragraph re-emphasizing the choice of x_C, x_qC and x_QC just after Equation (99); these terms do not have to the same leading terms in T^2.	2	1	The results obtained seem reasonable, although I have no time to verify all the derivations.			atoms4020014_perova	0
+Several additional paragraphs have been added near the figures to help explain the plots appearing in these figures. For the stationary atom case, the limits on y_{m0} are determined from inequality (106) and it is indeed possible for y_{m0} to be less than y_{min}. The manner in which we arrived at the regions of validity of the approximations is now described in more detail.	2	1	Authors’ response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			atoms4020014_perova	0
+In the article we obtained results that are specific for the parameters of the atom interferometer and test mass’ shape and trajectory. We think for the each case one should generate one figure 3 to answer all questions regarding the validity and role of different parts of the phase and approximate expressions for them. Instead of using a lot of dashed curves, we decided to use a panel of figures. Although each plot is difficult to read at “normal” magnification, the online reader can easily magnify any plot when accessing the article in PDF format. We have added a note to this effect.	2	1	The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			atoms4020014_perova	0
+We added a schematic figure of the geometry of our problem.	2	1	I recommend accepting this manuscript, provided the following points are clarified:			atoms4020014_perova	0
+In the paragraph after Equation (30) we added a list of requirements that will insure that the signal does not depend critically on the initial atomic state phase space distribution. These requirements minimize the corrections arising from diffraction to which the referee refers 11.	2	1	Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+Difficulties related to stray interferometers exist and are important, but are not addressed in our paper, as the referee points out. We are assuming that the effects can be minimized in a fountain geometry using a variety of experimental techniques. In any event, a discussion of these effects is beyond what we are able to discuss in this article.	2	1	The results obtained seem reasonable, although I have no time to verify all the derivations.			atoms4020014_perova	0
+The word “potential” has been changed to “acceleration” in the sentence after Equation (1).	2	1	The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+We know that this paper is “equation-rich”, but we have tried to provide some physical guidance for the reader.	2	1	The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			atoms4020014_perova	0
+A figure has been added to clarify the geometry of the problem. Moreover, additional discussion of the trajectories involved are now given in paragraphs that were added to help explain the plots given in Figures 2 and 3.	2	1	Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			atoms4020014_perova	0
+The wave function cannot be used when we know only some statistical properties of the initial phase space distribution.	2	1	The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+We thank the referee for pointing out this additional reference and have added it to the references.	2	1	Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			atoms4020014_perova	0
+As suggested by the reviewer, the non-pertinent references has been checked throughout the manuscript and unnecessary references are removed too.	2	1	Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			biom12060834_makarova	0
+As suggested by the reviewer, now the title include the production optimization too and the new title is “Optimization and characterization of a novel exopolysaccharide from Bacillus haynesii CamB6 for food applications”.	2	1	Comment 35: l. 500 : The chemical shift range is more than from 1 to 5.			biom12060834_makarova	0
+As suggested by the reviewer, the acceptability for food application has now been mentioned with appropriate refences (Nicolaus et al., 2010; Kambourova et al., 2018; Gongi et al., 2022) in the revised version of the manuscript.	2	1	Comment 14: l. 165: Table 2 where Table 1 expected Answer 14:			biom12060834_makarova	0
+According to reviewer´s suggestion, the language of the manuscript is thoroughly revised.	2	1	Comment 23: l. 340: “source” Answer 23: Suggestion has been included in the revised version of the manuscript Comment 24: Figure 2: No x-axis label, Units not specified Answer 24:			biom12060834_makarova	0
+As suggested by the reviewer, the consistent of the word had been checked throughout.	2	1	Comment 5: English language should be revised thoroughly.			biom12060834_makarova	0
+Constant abbreviation had been checked throughout in the revised version of the manuscript.	2	1	Fourier transform infrared spectroscopy (FTIR) and NMR spectroscopy data can only be regarded as preliminary.			biom12060834_makarova	0
+Accoding to the suggestion of the reviewer, Table 1 is revised and both coded – noncoded values are now included.	2	1	A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll.			biom12060834_makarova	0
+Both the tables (table 1 and table 2) have been placed in the proper place where it expected in the revised manuscript.	2	1	Comment 43: l. 711: “to be significantly” Answer 43: Suggestion has been included in the revised version of the manuscript Comment 44: l. 725: “possibility of future” Answer 44: Suggestion has been included in the revised version of the manuscript			biom12060834_makarova	0
+The suggested correction has been done in the revised version of the manuscript	2	1	The studied EPS showed promising properties to use in the food industry.			biom12060834_makarova	0
+The corrected has been made now.	2	1	See file Comments for author File: Comments.pdf			biom12060834_makarova	0
+The revision has been made as suggested.	2	1	Tick label values for glucose and yeast extract are different.			biom12060834_makarova	0
+as suggested by the reviewer, PEG has been defined in the revised version of the manuscript	2	1	Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.			biom12060834_makarova	0
+The volume is now added in the revised version of the manuscript.	2	1	Answer 1: To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript.			biom12060834_makarova	0
+Suggestion has been included in the revised version of the manuscript throughout.	2	1	Units are not specified on the axes labels or in the legend.			biom12060834_makarova	0
+This to kindly mention to the reviewer, the map is the collection site of the EPS producing Bacillus haynesii CamB6. This is a first such report from this collection site, the map is necessary to keep in the manuscript.	2	1	Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.			biom12060834_makarova	0
+Figure 2 has been modified as suggested by the reviewer.	2	1	This EPS was carefully purified according to standard purification protocols for EPS.			biom12060834_makarova	0
+As suggested by the reviewer, Figure 3 is now modified with specified units of the axes and also in the figure legend.	2	1	It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.			biom12060834_makarova	0
+All the Suggestion has been included in the figure legend of revised version of the manuscript	2	1	The authors added a COSY spectrum but did not analyze it.			biom12060834_makarova	0
+To avoid the redundancy of the figures as suggested by the reviewer, figure 4 is removed from the revised version of the manuscript.	2	1	Comment 7: Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.			biom12060834_makarova	0
+As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript	2	1	How can the authors ascertain that mannose does not come from mannans in yeast extract?			biom12060834_makarova	0
+This has been checked now and revised.	2	1	Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_makarova	0
+We performed 2D-NMR analysis , which allows us to determine exactly the α-Manp linkage, which is the major sugar component, and the β- structure of Galp and Glucp.	2	1	Manuscript biomolecules-1691308 In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			biom12060834_makarova	0
+This is a heteropolysaccharide composed of 3 different sugars as determined for HPLC, which is highly common in this kind of polysaccharides isolated from bacterial strains (10.3390/foods11020156)	2	1	→ Comment 1: The English language was improved, but there are still errors.			biom12060834_makarova	0
+There are several earlier report where different concentration of yeast extract is added to the culture media for optimized EPS production, and the resultant EPS also concisted of mannan.	2	1	Units are not specified on the axes labels or in the legend.			biom12060834_makarova	0
+Several 2D-NMR spectra were performed to confirm 1H and 13C chemical shift of EPS structure, and their analysis was added to the manuscript.	2	1	"Comment 3: Line 42: It is premature to include the term ""structure"" in keywords at this stage."			biom12060834_makarova	0
+It was corrected in manuscript	2	1	Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_makarova	0
+A new discussion of 1D and 2D NMR spectra was added and corrected.	2	1	Comment 14: l. 165: Table 2 where Table 1 expected Answer 14:			biom12060834_makarova	0
+According to the suggestion, section numbering has been checked thoroughly in the revised version of the manuscript.	2	1	Comment 22: Figure 1: What is the significance of the tick mark labels on the maps?			biom12060834_makarova	0
+As suggested by the reviewer, the necessary revision in done in the manuscript.	2	1	If the authors do not add methylation analysis and perform full NMR spectral analysis, the claims about structure determination should be removed from the manuscript.			biom12060834_makarova	0
+As suggested by the reviewer, the information is corrected and revised now.	2	1	), as well as two-dimensional NMR spectroscopy.			biom12060834_makarova	0
+Suggestion has been included in the revised version of the manuscript	2	1	Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			biom12060834_makarova	0
+As suggested by the reviewer, minor errors throughout the manuscript has been revised now.	4	1	TOCSY and HMBC are mentioned in methods, but no results are presented.			biom12060834_makarova	0
+int. We agree with some of the comments that you made. However, we would like to remind you that the main purpose of this manuscript is not the complete elucidation of the EPS chemical structure but also the optimization of the EPS production, and characterization of the EPS functional properties. At the same time, we provided different chemical properties like: sugar identification and its molar ratio, molecular weight, functional group analysis and thermal stability of the EPS, and this journal has recently published other papers with similar or less chemical information on the EPS, for instance: Kuo, H.-C.; Liu, Y.-W.; Lum, C.-C.; Hsu, K.-D.; Lin, S.-P.; Hsieh, C.-W.; Lin, H.-W.; Lu, T.-Y. ; Cheng, K.- C. Ganoderma formosanum Exopolysaccharides Inhibit Tumor Growth via Immunomodulation. Int. J. Mol. Sci. 2021, 22, 11251. https://doi.org/10.3390/ijms222011251 Fetsiukh, A.; Conrad, J.; Bergquist, J.; Timmusk, S. Silica Particles Trigger the Exopolysaccharide Production of Harsh Environment Isolates of Growth-Promoting Rhizobacteria and Increase Their Ability to Enhance Wheat Biomass in Drought-Stressed Soils. Int. J. Mol. Sci. 2021, 22, 6201. https://doi.org/10.3390/ijms22126201.	4	1	Author Response Please see the attachment for the responses.			biom12060834_makarova	0
+On the other side, we performed the 1D and 2D - NMR analysis as complementary information to the other techniques in the present work. Such analysis was performed to try to replace the traditional acid hydrolysis, methylation/acetylation and GC-MS used in glycoside analysis. On the opposite to NMR, we do not have access to this analysis, and we could not find it as a service.	4	1	Comment 4: Line 526-527: The proposal should be reformulated.			biom12060834_makarova	0
+The HPLC and NMR results from the present work confirmed the heterogeneity of the EPS (3 different sugar moieties with different molar ratios), which agrees with the other reported mannan polysaccharides. The 1H and 13C NMR chemical shifts are comparable with literature reports of other similar EPS or oligosaccharides, where complex structures are identified by NMR too (Casillo et al., 2021; Chatterjee et al; 2018). Besides, our 2D-NMR analysis supported EPS heterogeneity, identifying different types of monomer linkages.	4	1	They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			biom12060834_makarova	0
+Your mention of the non-uniform peak intensity confirming the three different sugar units found. This is because the signal intensity of NMR peaks is related to an -H abundance. Different monomer ratios are responsible for this variation and support our findings. Also, the 10 H1/H2 cross peaks observed in the COSY spectra supported our findings of EPS heterogeneity, given the following facts: 1) We found 3 different types of sugars by HPLC. 2) We identified by NMR at least 2 types of linkages α 1-2 Man-Man and alfa 1-4 Man-Man monosaccharides. 3) There are other 2 sugars for which the β- type of linkage was identified but not the exact sugar position. 4) Anomeric protons shifts from units at the reducing end of polysaccharide differs from the one in another position. In conclusion, there are different linkage possibilities (plus already identified) to different monosaccharides in different ring positions. This gives rise to a multiplicity of signals (not fully identified in this work), which could produce different H1/H2 correlations.	4	1	In addition, the yeast mannan have β-(1→4) linkage and the one found in this work have α-linkage type.			biom12060834_makarova	0
+About the COSY spectrum discussion, only a minor and partial discussion is presented that can be found below the Scheme in manuscript: “In the COSY spectrum, it was found its α-configuration by low-field H-1 signal at δ 5.34 and correlated with H-2 (δ 4.0), H-6 (δ 3.98 and 3.85), and H-5 (δ 4.12) (Speciale et al., 2022).”	4	1	Response 2: Dear reviewer, thanks for your valuable comments for improving the quality of this paper.			biom12060834_makarova	0
+Regarding the TOCSY and HMBC mentioned in the methods, they are now deleted from the manuscript, as suggested.	4	1	The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_makarova	0
+Finally, we kindly request the reviewer to consider that this is the first approach to studying a Chilean hot spring EPS isolation and its functional characterization. We also mention that further work to fully reveal this EPS structure will be necessary using higher resolution NMR spectra combined with chemical techniques. This work has given partial structural elucidation, which can be considered a previous background for other research on this topic.	4	1	The evidences for the partial structure given are not convincing.			biom12060834_makarova	0
+To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript. In short, they allow to clearly identify the D-mannose α-(1→2) and α-(1→4) linkages in EPS. These results suggest they are the prevailing units of the EPS backbone making it a branched one. Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.	2	1	There are inconsistencies in singular- plural concordance between subject and verb as well as noun and pronoun.			biom12060834_makarova	0
+According to the composition of EPS no metal was found to be attached with the structure of purified EPS. Likewise, in this study the monomeric composition was analysed to demonstrate the structure of purified EPS, where no metal was found to be attached with the chemical structure of it. This is also mentioned and referred in the revised version of the manuscript.	2	1	According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			biom12060834_makarova	0
+As suggested by the reviewer, this keyword is removed from the revised version of the manuscript.	2	1	The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			biom12060834_makarova	0
+Thank you so much dear reviewer for mentioning this po The correction has been made now in the revised version of the manuscript.	2	1	433, 434, 454, 458, and 507 : First section 3.6 (and its subsections) should be section 3.5.			biom12060834_makarova	0
+As suggested by the reviewer, the non-pertinent references has been checked throughout the manuscript and unnecessary references are removed too.	2	1	(I was referring to peak intensities that are not uniform.			biom12060834_perova	0
+As suggested by the reviewer, now the title include the production optimization too and the new title is “Optimization and characterization of a novel exopolysaccharide from Bacillus haynesii CamB6 for food applications”.	2	1	How can the authors ascertain that mannose does not come from mannans in yeast extract?			biom12060834_perova	0
+As suggested by the reviewer, the acceptability for food application has now been mentioned with appropriate refences (Nicolaus et al., 2010; Kambourova et al., 2018; Gongi et al., 2022) in the revised version of the manuscript.	2	1	Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+According to reviewer´s suggestion, the language of the manuscript is thoroughly revised.	2	1	Comment 16: l. 177: “βi” instead of “β0” in second term?			biom12060834_perova	0
+As suggested by the reviewer, the consistent of the word had been checked throughout.	2	1	Comment 15: l. 169-170: “thirty” instead of “thirteen”?			biom12060834_perova	0
+Constant abbreviation had been checked throughout in the revised version of the manuscript.	2	1	(I was referring to the chemical method by methylation.)			biom12060834_perova	0
+Accoding to the suggestion of the reviewer, Table 1 is revised and both coded – noncoded values are now included.	2	1	They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			biom12060834_perova	0
+Both the tables (table 1 and table 2) have been placed in the proper place where it expected in the revised manuscript.	2	1	Tick label values for glucose and yeast extract are different.			biom12060834_perova	0
+The suggested correction has been done in the revised version of the manuscript	2	1	Comment 32: l. 480: Why was linkage analysis not performed?			biom12060834_perova	0
+The corrected has been made now.	2	1	), as well as two-dimensional NMR spectroscopy.			biom12060834_perova	0
+The revision has been made as suggested.	2	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit.			biom12060834_perova	0
+as suggested by the reviewer, PEG has been defined in the revised version of the manuscript	2	1	Comment 2: The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.			biom12060834_perova	0
+The volume is now added in the revised version of the manuscript.	2	1	Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			biom12060834_perova	0
+Suggestion has been included in the revised version of the manuscript throughout.	2	1	"Comment 3: Line 42: It is premature to include the term ""structure"" in keywords at this stage."			biom12060834_perova	0
+This to kindly mention to the reviewer, the map is the collection site of the EPS producing Bacillus haynesii CamB6. This is a first such report from this collection site, the map is necessary to keep in the manuscript.	2	1	Chemical shift similarity is not sufficient to identify monosaccharide and linkage in a polysaccharide repeating unit.			biom12060834_perova	0
+Figure 2 has been modified as suggested by the reviewer.	2	1	Comment 4: Line 526-527: The proposal should be reformulated.			biom12060834_perova	0
+As suggested by the reviewer, Figure 3 is now modified with specified units of the axes and also in the figure legend.	2	1	Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			biom12060834_perova	0
+All the Suggestion has been included in the figure legend of revised version of the manuscript	2	1	), as well as two-dimensional NMR spectroscopy.			biom12060834_perova	0
+To avoid the redundancy of the figures as suggested by the reviewer, figure 4 is removed from the revised version of the manuscript.	2	1	Manuscript biomolecules-1691308 In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			biom12060834_perova	0
+As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript	2	1	According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			biom12060834_perova	0
+This has been checked now and revised.	2	1	481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			biom12060834_perova	0
+We performed 2D-NMR analysis , which allows us to determine exactly the α-Manp linkage, which is the major sugar component, and the β- structure of Galp and Glucp.	2	1	→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			biom12060834_perova	0
+This is a heteropolysaccharide composed of 3 different sugars as determined for HPLC, which is highly common in this kind of polysaccharides isolated from bacterial strains (10.3390/foods11020156)	2	1	Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+There are several earlier report where different concentration of yeast extract is added to the culture media for optimized EPS production, and the resultant EPS also concisted of mannan. However, there is no report that the mannan came from yeast extract (DOI: 10.1016/j.ijbiomac.2019.09.139, DOI: 10.1023/B:WIBI.0000033068.45655.2a). The same applies for our study too. In addition, the yeast mannan have β-(1→4) linkage and the one found in this work have α-linkage type. This allows us to conclude they are different polysaccharides.	2	1	Figure 6: Bacterial EPSs are normally composed of repeating units.			biom12060834_perova	0
+Several 2D-NMR spectra were performed to confirm 1H and 13C chemical shift of EPS structure, and their analysis was added to the manuscript.	2	1	See file Comments for author File: Comments.pdf			biom12060834_perova	0
+It was corrected in manuscript	2	1	See file Comments for author File: Comments.pdf			biom12060834_perova	0
+A new discussion of 1D and 2D NMR spectra was added and corrected.	2	1	According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.			biom12060834_perova	0
+According to the suggestion, section numbering has been checked thoroughly in the revised version of the manuscript.	2	1	Comment 27: l. 414, Delete “(“before “C. pH”. ll. 416 and 432 , Add “C.” before “pH=” . ll. 417 and 432 , “30.0” instead of “3.0” ?			biom12060834_perova	0
+As suggested by the reviewer, the necessary revision in done in the manuscript	2	1	In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			biom12060834_perova	0
+As suggested by the reviewer, the information is corrected and revised now	2	1	Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			biom12060834_perova	0
+Suggestion has been included in the revised version of the manuscript	2	1	Answer 29: As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript Comment 30: l. 442: “C), whereas the” Answer 30: Suggestion has been included in the revised version of the manuscript Comment 31: l. 471, 600, 678, and 718: Reference format is different; corresponding references could be missing in the list.			biom12060834_perova	0
+As suggested by the reviewer, minor errors throughout the manuscript has been revised now.	4	1	The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_perova	0
+Dear reviewer, thanks for your valuable comments for improving the quality of this paper. We will answer in the following paragraphs because all comments were related to the same topic.	4	1	Comment 23: l. 340: “source” Answer 23: Suggestion has been included in the revised version of the manuscript Comment 24: Figure 2: No x-axis label, Units not specified Answer 24:			biom12060834_perova	0
+We agree with some of the comments that you made. However, we would like to remind you that the main purpose of this manuscript is not the complete elucidation of the EPS chemical structure but also the optimization of the EPS production, and characterization of the EPS functional properties. At the same time, we provided different chemical properties like: sugar identification and its molar ratio, molecular weight, functional group analysis and thermal stability of the EPS, and this journal has recently published other papers with similar or less chemical information on the EPS, for instance: Kuo, H.-C.; Liu, Y.-W.; Lum, C.-C.; Hsu, K.-D.; Lin, S.-P.; Hsieh, C.-W.; Lin, H.-W.; Lu, T.-Y.; Cheng, K.- C. Ganoderma formosanum Exopolysaccharides Inhibit Tumor Growth via Immunomodulation. Int. J. Mol. Sci. 2021, 22, 11251. https://doi.org/10.3390/ijms222011251 Fetsiukh, A.; Conrad, J.; Bergquist, J.; Timmusk, S. Silica Particles Trigger the Exopolysaccharide Production of Harsh Environment Isolates of Growth-Promoting Rhizobacteria and Increase Their Ability to Enhance Wheat Biomass in Drought-Stressed Soils. Int. J. Mol. Sci. 2021, 22, 6201. https://doi.org/10.3390/ijms22126201.	4	1	Units are not specified on the axes labels or in the legend.			biom12060834_perova	0
+On the other side, we performed the 1D and 2D - NMR analysis as complementary information to the other techniques in the present work. Such analysis was performed to try to replace the traditional acid hydrolysis, methylation/acetylation and GC-MS used in glycoside analysis. On the opposite to NMR, we do not have access to this analysis, and we could not find it as a service.	4	1	According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			biom12060834_perova	0
+The HPLC and NMR results from the present work confirmed the heterogeneity of the EPS (3 different sugar moieties with different molar ratios), which agrees with the other reported mannan polysaccharides. The 1H and 13C NMR chemical shifts are comparable with literature reports of other similar EPS or oligosaccharides, where complex structures are identified by NMR too (Casillo et al., 2021; Chatterjee et al; 2018). Besides, our 2D-NMR analysis supported EPS heterogeneity, identifying different types of monomer linkages. Casillo, A., Fabozzi, A., Russo Krauss, I., Parrilli, E., Biggs, C. I., Gibson, M. I., … Corsaro, M. M. (2021). Physicochemical Approach to Understanding the Structure, Conformation, and Activity of Mannan Polysaccharides. Biomacromolecules, 22(4), 1445–1457. doi:10.1021/acs.biomac.0c01659. Chatterjee, S., Mukhopadhyay, S. K., Gauri, S. S., & Dey, S. (2018). Sphingobactan, a new α-mannan exopolysaccharide from Arctic Sphingobacterium sp. IITKGP-BTPF3 capable of biological response modification. International Immunopharmacology, 60, 84–95. doi:10.1016/j.intimp.2018.04.039 •	4	1	They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			biom12060834_perova	0
+Your mention of the non-uniform peak intensity confirming the three different sugar units found. This is because the signal intensity of NMR peaks is related to an -H abundance. Different monomer ratios are responsible for this variation and support our findings. Also, the 10 H1/H2 cross peaks observed in the COSY spectra supported our findings of EPS heterogeneity, given the following facts: 1) We found 3 different types of sugars by HPLC. 2) We identified by NMR at least 2 types of linkages α 1-2 Man-Man and alfa 1-4 Man-Man monosaccharides. 3) There are other 2 sugars for which the β- type of linkage was identified but not the exact sugar position. 4) Anomeric protons shifts from units at the reducing end of polysaccharide differs from the one in another position. In conclusion, there are different linkage possibilities (plus already identified) to different monosaccharides in different ring positions. This gives rise to a multiplicity of signals (not fully identified in this work), which could produce different H1/H2 correlations.	4	1	Tick label values for glucose and yeast extract are different.			biom12060834_perova	0
+About the COSY spectrum discussion, only a minor and partial discussion is presented that can be found below the Scheme in manuscript: “In the COSY spectrum, it was found its α-configuration by low-field H-1 signal at δ 5.34 and correlated with H-2 (δ 4.0), H-6 (δ 3.98 and 3.85), and H-5 (δ 4.12) (Speciale et al., 2022).”	4	1	They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			biom12060834_perova	0
+Regarding the TOCSY and HMBC mentioned in the methods, they are now deleted from the manuscript, as suggested.	4	1	These results suggest they are the prevailing units of the EPS backbone making it a branched one.			biom12060834_perova	0
+Finally, we kindly request the reviewer to consider that this is the first approach to studying a Chilean hot spring EPS isolation and its functional characterization. We also mention that further work to fully reveal this EPS structure will be necessary using higher resolution NMR spectra combined with chemical techniques. This work has given partial structural elucidation, which can be considered a previous background for other research on this topic.	4	1	Fourier transform infrared spectroscopy (FTIR) and NMR spectroscopy data can only be regarded as preliminary.			biom12060834_perova	0
+To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript. In short, they allow to clearly identify the D-mannose α-(1→2) and α-(1→4) linkages in EPS. These results suggest they are the prevailing units of the EPS backbone making it a branched one. Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.	2	1	Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			biom12060834_perova	0
+According to the composition of EPS no metal was found to be attached with the structure of purified EPS. Likewise, in this study the monomeric composition was analysed to demonstrate the structure of purified EPS, where no metal was found to be attached with the chemical structure of it. This is also mentioned and referred in the revised version of the manuscript.	2	1	Manuscript biomolecules-1691308 In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			biom12060834_perova	0
+As suggested by the reviewer, this keyword is removed from the revised version of the manuscript.	2	1	These results suggest they are the prevailing units of the EPS backbone making it a branched one.			biom12060834_perova	0
+Thank you so much dear reviewer for mentioning this point. The correction has been made now in the revised version of the manuscript.	2	1	Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+This is not the case; HSA and the other fluids tested as inductors were added to the growth medium (LB) medium, and the cells were allowed to grow in these conditions for 5 h at 37ºC. After that, the cells were collected, and RNA was extracted. Since HSA does not reach the cytosol, at least not in an intact manner, hydrolysis seems not possible during cell growth.	2	1	If we understand correctly, the reviewer’s argument, i.e., a reduction in RNA when the cells were cultured in the presence of HSA may be due to the HSA’s RNA-hydrolyzing activity, requires that HSA contacts the RNA that will be hydrolyzed.			biomedicines10030600_makarova	0
+It is also worth mentioning that the presence of HSA in the growth medium produced a reduction only in some mRNA species. These results speak against an unspecific hydrolyzing effect.	2	1	Therefore, the RNA is intracellular at all moments during the experiment, and the HSA stays in the milieu.			biomedicines10030600_makarova	0
+We also want to underscore that all RNA samples were checked after extraction. Agarose gels electrophoresis confirmed the integrity of RNA and lack of DNA contamination.	2	1	The same cDNA amount for every condition was used to perform qRT-PCR assays.			biomedicines10030600_makarova	0
+Finally, the article provided by the reviewer describes HSA hydrolysis of extracellular, not intracellular, RNA.	2	1	We would prefer to include those results in the upcoming publication.			biomedicines10030600_makarova	0
+A follow-up project that will be submitted for publication in the near future extends the studies of the effects produced by human fluids, and their components will include the experiments demonstrating a bactericidal effect of cefiderocol.	2	1	The authors extensively study the effects of human serum, human serum albumin, or human pleural fluid on Cefiderocol sensitivity of a panel of A. baumanii strains.			biomedicines10030600_makarova	0
+We thank the reviewer for their suggestion and have now computed AMPA-NMDA ratios and included a phrase in lines 424-425 that summarises the statistics for the comparison of AMPA-NMDA ratios between the GluN2B variants (WT and mutants).	2	1	3) A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion 4) A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.			brainsci12060789_makarova	0
+We thank the reviewer for this excellent suggestion. We have now measured the decay time constants for the NMDA-EPSCs before and after TCN-201. The graph of the data is shown in a new figure in the supplement, Fig. S4. The finding is insightful, and so we have added a paragraph relating to it in the results section, lines 520-536.	2	1	Nonetheless, in the presence of native GluN2A, molecular replacement of native GluN2B with GoF and LoF GluN2B mutants all led to similar functional incorporation into synaptic receptors, more rapidly decaying NMDA-EPSCs and greater inhibition by TCN-201, a selective antagonist of GluN2A-containing NMDA receptors.			brainsci12060789_makarova	0
+In answer to the last comment here from the reviewer, the time scale in Fig. 3A is correct – the examples chosen are representative of the group data in terms of the amount of block by TCN-201, but these examples all happen to have faster decays than the examples shown in Figures 1 and 2. In any case, to be more consistent with the formatting in some of the other figures, we have lengthened the scale bar and increased the time proportionally.	2	1	Author Response Thank you to the reviewer for their comments and helpful suggestions.			brainsci12060789_makarova	0
+This is an excellent idea. We have added an additional (final) main text figure, Fig. 7, illustrating how the molecular defects could converge on similar NMDA-EPSCs. We have explained the model in the accompanying figure legend with reference to supporting evidence in other figures.	2	1	The abstract now reads as follows: GRIN2B mutations are rare but often associated with patients having severe neurodevelopmental disorders, including a varying range of symptoms such as intellectual disability, developmental delay and epilepsy.			brainsci12060789_makarova	0
+We have added further discussion relating to the pathogenesis of neurodevelopmental disorders arising from GRIN2B mutations in lines 765-771 of the manuscript.	2	1	In the absence of both native GluN2A and GluN2B subunits, functional incorporation into synaptic NMDA receptors was attenuated for GoF mutants, or almost eliminated for LoF GluN2B mutants.			brainsci12060789_makarova	0
+It is hard to know exactly why the measurements of the internal control, untransfected neuron appear to vary between the experimental groups. While an effort was made to try and evoke synaptic responses with a consistent stimulus voltage setting, the effectiveness of the stimulus for evoking an NMDA-EPSC depends on, among other things, the condition of the stimulating electrode (e.g. resistance, stray capacitance, bubbles etc). It was rarely practical to record all mutant conditions in the same experiment, so variation in the stimulating electrode condition over the course of this series of experiments could potentially lead to some apparent differences between untransfected neurons with respect to their mean NMDA-EPSC peak amplitude and charge transfer. What this does serve to illustrate though is how important it was for us to use untransfected neurons as an internal control.	2	1	Marwa Elmasri and colleagues report that several GoF and LoF mutations in GRIN2B influence NMDA functions.			brainsci12060789_makarova	0
+The supplementary figures were within the same PDF so we are not sure why the author had trouble gaining accessing to the figures. Assuming the reviewer had trouble understanding the contents of the supplement, we have tried to simplify somewhat the text in the supplementary figure legends.	2	1	The abstract now reads as follows: GRIN2B mutations are rare but often associated with patients having severe neurodevelopmental disorders, including a varying range of symptoms such as intellectual disability, developmental delay and epilepsy.			brainsci12060789_makarova	0
+We thank the reviewer for this comment. Indeed, there were many long sentences and the abstract did not capture all of our findings. We have rewritten the abstract to accommodate the reviewer’s suggestions. The abstract is now 311 words long (almost 40 words shorter than the original abstract), has shorter sentences, and includes some summary of the findings reported in all the main figures.	2	1	The abstract now reads as follows: GRIN2B mutations are rare but often associated with patients having severe neurodevelopmental disorders, including a varying range of symptoms such as intellectual disability, developmental delay and epilepsy.			brainsci12060789_makarova	0
+We thank the reviewer for their suggestion and have now computed AMPA-NMDA ratios and included a phrase in lines 424-425 that summarises the statistics for the comparison of AMPA-NMDA ratios between the GluN2B variants (WT and mutants).	2	1	Please find below our point-by-point response.			brainsci12060789_perova	0
+We thank the reviewer for this excellent suggestion. We have now measured the decay time constants for the NMDA-EPSCs before and after TCN-201. The graph of the data is shown in a new figure in the supplement, Fig. S4. The finding is insightful, and so we have added a paragraph relating to it in the results section, lines 520-536. In answer to the last comment here from the reviewer, the time scale in Fig. 3A is correct – the examples chosen are representative of the group data in terms of the amount of block by TCN-201, but these examples all happen to have faster decays than the examples shown in Figures 1 and 2. In any case, to be more consistent with the formatting in some of the other figures, we have lengthened the scale bar and increased the time proportionally.	2	1	The results presented are convincing and I only have a few minor comments.			brainsci12060789_perova	0
+This is an excellent idea. We have added an additional (final) main text figure, Fig. 7, illustrating how the molecular defects could converge on similar NMDA-EPSCs. We have explained the model in the accompanying figure legend with reference to supporting evidence in other figures.	2	1	Particularly, GluN2B GoF and LoF mutants show similar phenotypes in the presence of GluN1/2A/2B triheteromers, while the LoF mutation, C456Y, shows much less rescue effect than GoF mutation in the absence of both GluN2A and GluN2B.			brainsci12060789_perova	0
+We have added further discussion relating to the pathogenesis of neurodevelopmental disorders arising from GRIN2B mutations in lines 765-771 of the manuscript.	2	1	3) A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion 4) A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.			brainsci12060789_perova	0
+It is hard to know exactly why the measurements of the internal control, untransfected neuron appear to vary between the experimental groups. While an effort was made to try and evoke synaptic responses with a consistent stimulus voltage setting, the effectiveness of the stimulus for evoking an NMDA-EPSC depends on, among other things, the condition of the stimulating electrode (e.g. resistance, stray capacitance, bubbles etc). It was rarely practical to record all mutant conditions in the same experiment, so variation in the stimulating electrode condition over the course of this series of experiments could potentially lead to some apparent differences between untransfected neurons with respect to their mean NMDA-EPSC peak amplitude and charge transfer. What this does serve to illustrate though is how important it was for us to use untransfected neurons as an internal control.	2	1	The results presented are convincing and I only have a few minor comments.			brainsci12060789_perova	0
+The supplementary figures were within the same PDF so we are not sure why the author had trouble gaining accessing to the figures. Assuming the reviewer had trouble understanding the contents of the supplement, we have tried to simplify somewhat the text in the supplementary figure legends.	2	1	There are also a number of sentences that are so complicated for easy read.			brainsci12060789_perova	0
+We thank the reviewer for this comment. Indeed, there were many long sentences and the abstract did not capture all of our findings. We have rewritten the abstract to accommodate the reviewer’s suggestions. The abstract is now 311 words long (almost 40 words shorter than the original abstract), has shorter sentences, and includes some summary of the findings reported in all the main figures. The abstract now reads as follows: GRIN2B mutations are rare but often associated with patients having severe neurodevelopmental disorders, including a varying range of symptoms such as intellectual disability, developmental delay and epilepsy. Patient symptoms likely arise from mutations disturbing the role that the encoded NMDA receptor subunit, GluN2B, plays at neuronal connections in the developing nervous system. In this study, we have investigated the cell-autonomous effects of putative gain- (GoF) and loss-of-function (LoF) missense GRIN2B mutations on excitatory synapses onto CA1 pyramidal neurons in organotypic hippocampal slices. In the absence of both native GluN2A and GluN2B subunits, functional incorporation into synaptic NMDA receptors was attenuated for GoF mutants, or almost eliminated for LoF GluN2B mutants. NMDA receptor-mediated excitatory postsynaptic currents (NMDA-EPSCs) from synaptic GoF GluN1/2B receptors had prolonged decays consistent with their functional classification. Nonetheless, in the presence of native GluN2A, molecular replacement of native GluN2B with GoF and LoF GluN2B mutants all led to similar functional incorporation into synaptic receptors, more rapidly decaying NMDA-EPSCs and greater inhibition by TCN-201, a selective antagonist of GluN2A-containing NMDA receptors. Mechanistic insight was gained from experiments in HEK293T cells, which revealed that GluN2B GoF mutants slowed deactivation in diheteromeric GluN1/2B, but not triheteromeric GluN1/2A/2B receptors. We also show that a disease-associated missense mutation, which severely affects surface expression, causes opposing effects on NMDA-EPSC decay and charge transfer when introduced into GluN2A or GluN2B. Finally, we show that having a single null Grin2b allele has only a modest effect on NMDA-EPSC decay kinetics. Our results demonstrate that functional incorporation of GoF and LoF GluN2B mutants into synaptic receptors and the effects on EPSC decay times are highly dependent on the presence of triheteromeric GluN1/2A/2B NMDA receptors, thereby influencing the functional classification of NMDA receptor variants as GoF or LoF mutations. These findings highlight the complexity of interpreting effects of disease-causing NMDA receptor missense mutations in the context of neuronal function.	2	1	Marwa Elmasri and colleagues report that several GoF and LoF mutations in GRIN2B influence NMDA functions.			brainsci12060789_perova	0
+We concur that a brief description of the literature regarding EIH in FM is warranted and have added information to the discussion as follows: “Previous research regarding the effects of acute bouts of exercise on pain sensitivity in FM patients is largely equivocal. Some studies have demonstrated a hypoalgesic effect [5,24] while other studies show either no changes in pain perception or an exacerbation of pain [25–27].”	2	1	In paralell pain sensitivity to heat was decreased by exercise.			brainsci6010008_makarova	0
+With respect to the reviewer’s comment regarding the lack of EIH in controls, we have included some additional information to demonstrate the consistency of our findings with previous research. The passage now reads: “However, it appears that the exercise bout as prescribed was not a sufficient stimulus to induce a decrease in pain sensitivity in our pain-free controls. This is consistent with previous literature regarding the effects of acute exercise on pain in healthy controls, which typically finds that higher intensity exercise is necessary to elicit a hypoalgesic response [33].” Additionally, we want to again mention that our study was not primarily designed to induce EIH in controls; it was designed to examine brain responses to pain following exercise and thus we wanted to ensure that FM patients could complete the exercise bout. Exploring brain responses associated with EIH in healthy individuals would be an interesting future direction.	2	1	Author Response We would like to thank the reviewer for their positive and critical review of our work.			brainsci6010008_makarova	0
+With respect to the reviewer’s comment on generalizability, we agree that our exclusionary criteria were stringent and have added the following sentence to our limitations section. “Further, we excluded individuals with diagnosed mental health conditions and those who were taking medications that could impact pain or the interpretation of brain responses. As such our results may not apply to FM patients with comorbid conditions.”	2	1	Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			brainsci6010008_makarova	0
+Lastly, with respect to the comment about controlling for physiological noise, the following statements have been added. “Though the evidence is equivocal, it has been suggested that cardiovascular mechanisms may be involved in the hypoalgesic response to exercise [34] and thus statistically controlling for these effects may have influenced the interpretation of our results. However, without controlling physiological differences between EX and QR the neuroimaging data would have been very difficult to interpret.” I don’t see that the reference actually indicates studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	2	1	I will be happy to read the revised version of the manuscript.			brainsci6010008_makarova	0
+This sentence has been revised to better reflect the reference used.	2	1	We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_makarova	0
+We agree. We consulted with each physician with respect to type and dosage of medication, including whether this was a low, moderate or high dose. We also double-checked each medication and dose with the Physician’s Desk Reference (60th-65th editions). We have added the following information to the methods section “Medication information and dosage were supplied by the patient and their physician and dosage levels (low, moderate, high) were determined through both physician consultation and use of the Physician’s Desk Reference (65th edition).	2	1	However, in the latter case the authors need to address the issue of multiple correlations.			brainsci6010008_makarova	0
+This addition has been made.	2	1	I will be happy to read the revised version of the manuscript.			brainsci6010008_makarova	0
+The suggested change has been made.	2	1	I will be happy to read the revised version of the manuscript.			brainsci6010008_makarova	0
+Thank you for catching this omission. This was an oversight on our part and these effect sizes are now included on page 11.	2	1	We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_makarova	0
+These analyses were intended primarily for descriptive purposes. However, we recognize the need to control for multiple comparisons in order to reduce the risk of making Type I errors. Therefore, we created 3 families including 2 correlations each (one for each of the significant regions) and performed a Bonferroni correction, making the critical alpha level for significance 0.025. This has been clarified in the statistical analysis section.	2	1	I am uncertain why the authors only report the effect sizes of group differences in the first run.			brainsci6010008_makarova	0
+This change has been made.	2	1	Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_makarova	0
+This correction has been made.	2	1	Nine individuals in each group were indicated in the neuroimaging analysis, this should be indicated in the abstract.			brainsci6010008_makarova	0
+We agree that this would add valuable information to the discussion section and have added the paragraph shown below discussing previous work using neuroimaging to understand the effects of exercise on the brain.	2	1	Nine individuals in each group were indicated in the neuroimaging analysis, this should be indicated in the abstract.			brainsci6010008_makarova	0
+A growing number of studies have begun to employ neuroimaging methods to better understand the impact of exercise on the brain both longitudinally and acutely. For example, Smith and colleagues [37] conducted fMRI scans pre and post an exercise training program in older adults with mild cognitive impairment and found that exercise improved neural efficiency during cognitive tasks post-intervention. Structural MRI has also been used to show the neuroprotective effects of regular exercise in older adults with respect to preservation of brain volume[38]. In contrast to using neuroimaging to track changes in the brain over time, neuroimaging during and immediately following exercise presents some unique challenges due to artifacts associated with movement and the physiological underpinnings of many neuroimaging methods (e.g. BOLD response). EEG has been used most extensively to explore the effects of exercise on cortical activity [39]. PET and fMRI have also been used, though to a much lesser extent. For example, Boecker and colleagues used PET to demonstrate the effects of a long-distance run on opioid release in the brain and Janse Van Rensberg and colleagues used fMRI to examine brain responses to nicotine craving following 10 minutes of moderate intensity cycling. Our study adds to this important body of literature by using fMRI to show that an acute bout of moderate intensity exercise improved brain mechanisms underlying pain modulation in patients with chronic pain and further highlights the potential benefits of utilizing neuroimaging technology to better understand the more immediate effects of exercise on the human brain.	2	1	We have addressed each concern below and made changes in the manuscript accordingly.			brainsci6010008_makarova	0
+We concur that a brief description of the literature regarding EIH in FM is warranted and have added information to the discussion as follows: “Previous research regarding the effects of acute bouts of exercise on pain sensitivity in FM patients is largely equivocal. Some studies have demonstrated a hypoalgesic effect [5,24] while other studies show either no changes in pain perception or an exacerbation of pain [25–27].”	2	1	We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_perova	0
+With respect to the reviewer’s comment regarding the lack of EIH in controls, we have included some additional information to demonstrate the consistency of our findings with previous research. The passage now reads: “However, it appears that the exercise bout as prescribed was not a sufficient stimulus to induce a decrease in pain sensitivity in our pain-free controls. This is consistent with previous literature regarding the effects of acute exercise on pain in healthy controls, which typically finds that higher intensity exercise is necessary to elicit a hypoalgesic response [33].” Additionally, we want to again mention that our study was not primarily designed to induce EIH in controls; it was designed to examine brain responses to pain following exercise and thus we wanted to ensure that FM patients could complete the exercise bout. Exploring brain responses associated with EIH in healthy individuals would be an interesting future direction.	2	1	Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			brainsci6010008_perova	0
+With respect to the reviewer’s comment on generalizability, we agree that our exclusionary criteria were stringent and have added the following sentence to our limitations section. “Further, we excluded individuals with diagnosed mental health conditions and those who were taking medications that could impact pain or the interpretation of brain responses. As such our results may not apply to FM patients with comorbid conditions.”	2	1	The methodology is sound and the manuscript is well-written.			brainsci6010008_perova	0
+Lastly, with respect to the comment about controlling for physiological noise, the following statements have been added. “Though the evidence is equivocal, it has been suggested that cardiovascular mechanisms may be involved in the hypoalgesic response to exercise [34] and thus statistically controlling for these effects may have influenced the interpretation of our results. However, without controlling physiological differences between EX and QR the neuroimaging data would have been very difficult to interpret.” I don’t see that the reference actually indicates studies of activities of daily living so the authors should revise the sentence or add an appropriate reference.	2	1	In paralell pain sensitivity to heat was decreased by exercise.			brainsci6010008_perova	0
+This sentence has been revised to better reflect the reference used.	2	1	Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			brainsci6010008_perova	0
+We agree. We consulted with each physician with respect to type and dosage of medication, including whether this was a low, moderate or high dose. We also double-checked each medication and dose with the Physician’s Desk Reference (60th-65th editions). We have added the following information to the methods section “Medication information and dosage were supplied by the patient and their physician and dosage levels (low, moderate, high) were determined through both physician consultation and use of the Physician’s Desk Reference (65th edition).	2	1	The methodology is sound and the manuscript is well-written.			brainsci6010008_perova	0
+This addition has been made.	2	1	Changes in pain sensitivity after exercise vs. quiet rest were significantly correlated with changes in activity in the DLPFC (exercise vs. rest).			brainsci6010008_perova	0
+The suggested change has been made.	2	1	Author Response We would like to thank the reviewer for their positive and critical review of our work.			brainsci6010008_perova	0
+Thank you for catching this omission. This was an oversight on our part and these effect sizes are now included on page 11.	2	1	The time post-exercise before scanning was provided, but not the time post-quiet rest.			brainsci6010008_perova	0
+These analyses were intended primarily for descriptive purposes. However, we recognize the need to control for multiple comparisons in order to reduce the risk of making Type I errors. Therefore, we created 3 families including 2 correlations each (one for each of the significant regions) and performed a Bonferroni correction, making the critical alpha level for significance 0.025.	2	1	The discussion interprets the results as supporting evidence of exercise-induced hypoalgesia (EIH) in the patients with FM.			brainsci6010008_perova	0
+This has been clarified in the statistical analysis section.	2	1	In table 3, the subheading “Peak X, Y, X” needs correction.			brainsci6010008_perova	0
+This change has been made.	2	1	However, in the latter case the authors need to address the issue of multiple correlations.			brainsci6010008_perova	0
+This correction has been made.	2	1	Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			brainsci6010008_perova	0
+We agree that this would add valuable information to the discussion section and have added the paragraph shown below discussing previous work using neuroimaging to understand the effects of exercise on the brain.	2	1	This manuscript describes a cohort randomized crossover design comparing cutaneous heat pain ratings and brain activity during heat stimuli after conditions of quiet rest and cycling in patients with a physician-confirmed diagnosis of Fibromyalgia (FM) and age and sex matched controls.			brainsci6010008_perova	0
+A growing number of studies have begun to employ neuroimaging methods to better understand the impact of exercise on the brain both longitudinally and acutely. For example, Smith and colleagues [37] conducted fMRI scans pre and post an exercise training program in older adults with mild cognitive impairment and found that exercise improved neural efficiency during cognitive tasks post-intervention. Structural MRI has also been used to show the neuroprotective effects of regular exercise in older adults with respect to preservation of brain volume[38]. In contrast to using neuroimaging to track changes in the brain over time, neuroimaging during and immediately following exercise presents some unique challenges due to artifacts associated with movement and the physiological underpinnings of many neuroimaging methods (e.g. BOLD response). EEG has been used most extensively to explore the effects of exercise on cortical activity [39]. PET and fMRI have also been used, though to a much lesser extent. For example, Boecker and colleagues used PET to demonstrate the effects of a long-distance run on opioid release in the brain and Janse Van Rensberg and colleagues used fMRI to examine brain responses to nicotine craving following 10 minutes of moderate intensity cycling. Our study adds to this important body of literature by using fMRI to show that an acute bout of moderate intensity exercise improved brain mechanisms underlying pain modulation in patients with chronic pain and further highlights the potential benefits of utilizing neuroimaging technology to better understand the more immediate effects of exercise on the human brain.	2	1	A reference for the determination that a dosage of antidepressants was “high” should be provided.			brainsci6010008_perova	0
+In the revised manuscript, this was corrected as suggested.	2	1	The writing is clear, the analysis is sound, and the presentation is excellent with a few exceptions.			brainsci6010009_makarova	0
+Both cases and controls could have been diagnosed with autism, but it should have been a relative minority. Autism is relatively rare compared to a 314 diagnosis. This issue was added to the study limitations in the revised manuscript. “In the present study, neurodevelopmental diagnoses other than 314.xx were not examined among cases and controls. This limitation of the present study should have had a limited impact on the results observed because of the rarity of other neurodevelopmental diagnoses as compared to a 314.xx diagnosis, but future studies could further evaluate this phenomenon.” Comment 3.	2	1	These results are relevant and important as thimerosal is still included in global vaccines although they have largely been removed from childhood vaccine in the United States.			brainsci6010009_makarova	0
+We don’t have the exact number, but it is assumed to be a relatively small number.	2	1	In the exposed and control groups were there other related diseases such as Autism?			brainsci6010009_makarova	0
+In the revised manuscript, this was corrected with a better reference.	2	1	The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			brainsci6010009_makarova	0
+In the revised manuscript, this information was added. “This database is available to outside researchers after obtaining approval from the CDC and KP. Information regarding access to this database is at: http://www.cdc.gov/vaccinesafety/ensuringsafety/monitoring/vsd/accessing-data.html.”	2	1	On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			brainsci6010009_makarova	0
+Thank you.	2	1	This is a straightforward analysis of an important database with profound implications for public health.			brainsci6010009_makarova	0
+In the revised manuscript, this was corrected as suggested.	2	1	Can the authors address this point: is this database publicly available?			brainsci6010009_perova	0
+Both cases and controls could have been diagnosed with autism, but it should have been a relative minority. Autism is relatively rare compared to a 314 diagnosis. This issue was added to the study limitations in the revised manuscript. “In the present study, neurodevelopmental diagnoses other than 314.xx were not examined among cases and controls. This limitation of the present study should have had a limited impact on the results observed because of the rarity of other neurodevelopmental diagnoses as compared to a 314.xx diagnosis, but future studies could further evaluate this phenomenon.”	2	1	On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			brainsci6010009_perova	0
+We don’t have the exact number, but it is assumed to be a relatively small number.	2	1	The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			brainsci6010009_perova	0
+In the revised manuscript, this was corrected with a better reference.	2	1	On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			brainsci6010009_perova	0
+In the revised manuscript, this information was added. “This database is available to outside researchers after obtaining approval from the CDC and KP. Information regarding access to this database is at: http://www.cdc.gov/vaccinesafety/ensuringsafety/monitoring/vsd/accessing-data.html.”	2	1	"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."			brainsci6010009_perova	0
+Thank you.	2	1	The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			brainsci6010009_perova	0
+Thank you for the enthusiastic comments.	2	1	Round 1: and Author Response This manuscript reports on person-centered analysis of adolescent girls finding that they are distinguished by types of aggression, callous-unemotional traits, and by interpersonal relationships with peers and parents.			bs5040518_makarova	0
+This is now provided in the results section.	2	1	Round 1: Author Response to Reviewer 2 In my view, this manuscript will make a nice contribution to the literature.			bs5040518_makarova	0
+Thank you! I have now carefully read and edited the paper.	2	1	The study is strengthened by the person-centered approach to data analysis, by the large sample, by focusing on girls who have been much less studied than boys, and by the clear rationale for the study.			bs5040518_makarova	0
+True that we do not know the direction of effects. Here, we use predictor in the statistical sense not causal sense.	2	1	Overall, this is a nice paper that looks to make a contribution.			bs5040518_makarova	0
+We agree this would be useful but it was not our aim to look at interactions. We also believe interactions such as suggested are better done in longitudinal studies where one can examine moderators.	2	1	First, it is recommended that the authors provide more information about the selection of the cluster solution.			bs5040518_makarova	0
+We have attempted to be mindful of causal language and only use it where prior research indicates a direction.	2	1	Despite the use of a relatively large sample and of sound measures, this study falls short of making an important contribution to the current literature for several reasons.			bs5040518_makarova	0
+We agree this is a major flaw and we thank the reviewer for pointing out projection bias. We now include this in the limitations in the discussion.	2	1	That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth.			bs5040518_makarova	0
+We agree with the reviewer that there is no easy answer to this problem. We now include a discussion of this in the introduction and cite Lynam and colleagues on this exact dilemma. In their paper, they discuss the Perils of Partialing (p.4, line 15), which is what you do when you control for the overlap. That is, one may be removing the reliable aspect of the self-report measures of aggression (when they are highly correlated) and remaining with residual error.	2	1	Round 1: and Author Response This manuscript reports on person-centered analysis of adolescent girls finding that they are distinguished by types of aggression, callous-unemotional traits, and by interpersonal relationships with peers and parents.			bs5040518_makarova	0
+The reactive aggressive group did not differ from the combined group on peer delinquency. Thus, both were high on delinquency and peer delinquency. Also, we comment on the overlap between peer delinquency and self-report of delinquency in the results section (p.9, line 14).	2	1	For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			bs5040518_makarova	0
+We have carefully edited the paper.	2	1	Creating groups based on a cluster analysis may have created unnecessary problems.			bs5040518_makarova	0
+The manuscript has been edited for grammar problems and for clarity. We have also added Table 1 to assist with understanding our predictors and the broader factors they represent in this study.	2	1	Round 1: Author Response to Reviewer 3 First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression.			bs5040518_makarova	0
+Thank you for the enthusiastic comments.	2	1	Round 1: and Author Response The goal of this study was to distinguish subgroups of relationally aggressive females (i.e., reactively aggressive-only subgroup, reactively-and-proactively aggressive-combined subgroup, non-aggressive subgroup), on the basis of nine factors reflecting individual characteristics, peer-related variables and R3 parental control.			bs5040518_perova	0
+This is now provided in the results section.	2	1	Round 1: and Author Response This manuscript reports on person-centered analysis of adolescent girls finding that they are distinguished by types of aggression, callous-unemotional traits, and by interpersonal relationships with peers and parents.			bs5040518_perova	0
+Thank you! I have now carefully read and edited the paper.	2	1	Overall, this is a nice paper that looks to make a contribution.			bs5040518_perova	0
+True that we do not know the direction of effects. Here, we use predictor in the statistical sense not causal sense.	2	1	There are two areas to address to strengthen the paper a bit more.			bs5040518_perova	0
+We agree this would be useful but it was not our aim to look at interactions. We also believe interactions such as suggested are better done in longitudinal studies where one can examine moderators.	2	1	Round 1: and Author Response This manuscript reports on person-centered analysis of adolescent girls finding that they are distinguished by types of aggression, callous-unemotional traits, and by interpersonal relationships with peers and parents.			bs5040518_perova	0
+We have attempted to be mindful of causal language and only use it where prior research indicates a direction.	2	1	In my view, this manuscript will make a nice contribution to the literature.			bs5040518_perova	0
+We agree this is a major flaw and we thank the reviewer for pointing out projection bias. We now include this in the limitations in the discussion.	2	1	First, it is recommended that the authors provide more information about the selection of the cluster solution.			bs5040518_perova	0
+We agree with the reviewer that there is no easy answer to this problem. We now include a discussion of this in the introduction and cite Lynam and colleagues on this exact dilemma. In their paper, they discuss the Perils of Partialing (p.4, line 15), which is what you do when you control for the overlap. That is, one may be removing the reliable aspect of the self-report measures of aggression (when they are highly correlated) and remaining with residual error.	2	1	In my view, this manuscript will make a nice contribution to the literature.			bs5040518_perova	0
+The reactive aggressive group did not differ from the combined group on peer delinquency. Thus, both were high on delinquency and peer delinquency. Also, we comment on the overlap between peer delinquency and self-report of delinquency in the results section (p.9, line 14).	2	1	It would help the reader to have some information available as a way of arguing more strongly that the accepted solution (which does make theoretical and empirical sense) is the strongest grouping.			bs5040518_perova	0
+We have carefully edited the paper.	2	1	Round 1: Author Response to Reviewer 3 First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression.			bs5040518_perova	0
+The manuscript has been edited for grammar problems and for clarity.	2	1	For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			bs5040518_perova	0
+We have also added Table 1 to assist with understanding our predictors and the broader factors they represent in this study.	2	1	For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			bs5040518_perova	0
+Thank you for this advice. We have trimmed the paper to focus on two of the five elements of sexual subjectivity - only those that were expected to be most relevant to partnered sexual partner: entitlement to sexual partner pleasure and efficacy in achieving pleasure. This has substantially limited the analyses conducted. We provide a rationale for this (see the first paragraph on p.4) and have thoroughly revised the entire paper.	2	1	Limitations of the study include a sample of predominantly middle class, Caucasian students at one university and the possibility that students more interested in sex and relationships, and with more sexual experience, chose to participate.			bs6010004_makarova	0
+We have done this and have also substantially revised the entire Introduction (see new section starting on p. 3 titled Definition and measurement).	2	1	Therefore, the results of the current study may have some limited generalizability.			bs6010004_makarova	0
+We have corrected this sentence.	2	1	A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			bs6010004_makarova	0
+We have added to our theoretical explanation for why there would be gender differences (see the section titled Gender Differences in Sexual Behavior, Attitudes, and Sexual Subjectivity, p. 4).	2	1	Age was positively associated with T1 entitlement, and experience with a wider range of partnered sexual behaviors was concurrently associated with more entitlement and efficacy and was also associated with increased entitlement to partner pleasure and increased self-efficacy in achieving sexual pleasure at T2 relative to T1.			bs6010004_makarova	0
+We have substantially revised all sections of this paper to improve the flow and removed some headings.	2	1	Therefore, the results of the current study may have some limited generalizability.			bs6010004_makarova	0
+We have also added hypotheses (see p. 5-6)	2	1	Are behaviors in early adolescence (before age 16), for example, expected to continue to influence the trajectory of sexual subjectivity regardless of later sexual behavior?			bs6010004_makarova	0
+We have revised this section (The Current Study, p.5).	2	1	** We added a comment that data were collected at one Australian university to the abstract, and added a summary statement and note about limitations related to generalizability.			bs6010004_makarova	0
+We have moved this content on group difference to the first section of the Results section.	2	1	Author Response There might be some interesting data collected in the present study.			bs6010004_makarova	0
+Given our focus on only two element of sexual subjectivity, this was no longer an issue because items were the same on measures for males and females.	2	1	"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion."			bs6010004_makarova	0
+"We have added this information on p. 7 in the Procedure section. It reads: Approval from the university Human Research Ethics Committee was obtained prior to data collection. At Time 1 (T1), participants were approached at a university campus in Australia in the week before classes commenced (i.e., during orientation week) and asked to participate in a study ""About You and Your Relationships."" The front cover of the survey described the questions as focused on personal sexual and romantic experiences, and stressed the confidential nature of the survey."	2	1	A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			bs6010004_makarova	0
+Reducing the number of elements of sexual subjectivity has reduced the number of tests performed.	2	1	The paper would flow more smoothly if some transition sentences were included.			bs6010004_makarova	0
+We have added hypotheses to the section titled The Current Study on p. 5-6.	2	1	Age was positively associated with T1 entitlement, and experience with a wider range of partnered sexual behaviors was concurrently associated with more entitlement and efficacy and was also associated with increased entitlement to partner pleasure and increased self-efficacy in achieving sexual pleasure at T2 relative to T1.			bs6010004_makarova	0
+We have now used multiple imputation to maintain all 295 participants in all analyses.	2	1	**We did not keep an exact count of the number of students approached about the study, but we estimate that we approached about 375 students.			bs6010004_makarova	0
+We are sorry that we did not keep a tally of the number of students approached. We have added to the Limitations section of the Discussion about this (see p. 12). It reads, First, the participants in the study were predominantly middle class, Caucasian university students, residing in one region of Australia. There is also the possibility that individuals more interested in sex and relationships chose to participate. Therefore, the results of the current study may have some limited generalizability.	2	1	Therefore, the results of the current study may have some limited generalizability.			bs6010004_makarova	0
+We agree that this is very important. We have added the Introduction to expand on these points. Unfortunately we could not conduct these suggested analyses. We had very little change in sexual behavior given the way we measured it. Also, we did not have confidence in estimation of sexual behavior for those who did not participate at Wave 2, which could have increased the change. Thus, we have maintained our original analyses, but make this comment in the Discussion (see p. 12-13): Second, probably because the majority of the participants had already experienced coitus prior to the first data collection and because we measured types of sexual behavior and not frequency or some other aspect of behavior, there was high stability in sexual behavior over the one year of this study. Recent statistics show that the majority of Australian year 10-12 students (approximately 15-17 years) have engaged in some form of sexual behavior [12]. Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with change in sexual behavior, future studies may need to begin with a younger population. Recently, Hensel et al. [31] found that, in their adolescent participants, sexual self-concept had a bidirectional relationship with sexual experience, and the same may occur for sexual subjectivity. Future research could examine the possibility of bidirectional relationships between sexual subjectivity and sexual behavior. In addition, no previous study has examined the timing of pubertal development and sexual subjectivity. Evidence suggests that adolescents that mature earlier, compared to their peers, form romantic relationships earlier and experience sexual behaviors earlier [11,55]. In the current and past research, the link between age and sexual subjectivity has been relatively weak [34]. Substituting timing of pubertal status for age may provide more evidence regarding individual characteristics that influence differences in sexual subjectivity and its development over time.	2	1	Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with sexual behavior, future studies may need to begin with a younger population.			bs6010004_makarova	0
+We have trimmed the paper to focus on two of the five elements of sexual subjectivity and anticipated that these two elements would show different results. Thus, we have maintained the separate analyses of these two elements. Please see the added Hypotheses on p. 5-6.	2	1	Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.			bs6010004_makarova	0
+We have removed these analyses from the paper in an attempt to reduce the number of analyses and streamline the entire paper.	2	1	The first paragraph of the Participants section now reads: The participants were 295 adolescent and young adult men (n = 112) and women (n = 183) aged between 17 to 25 years (M = 19.5 years, SD = 1.9).			bs6010004_makarova	0
+Thank you for this suggestion. The reference has been added to the paper.	2	1	The authors rely too much on headings to transition between ideas.			bs6010004_makarova	0
+Thank you for this advice. We have trimmed the paper to focus on two of the five elements of sexual subjectivity - only those that were expected to be most relevant to partnered sexual partner: entitlement to sexual partner pleasure and efficacy in achieving pleasure. This has substantially limited the analyses conducted. We provide a rationale for this (see the first paragraph on p.4) and have thoroughly revised the entire paper.	2	1	**We also expanded the first paragraph of the Limitations to say: Although this study provided insight into how aspects of sexual subjectivity differed over one year in young men and women and uncovered associations of age and sexual behavior with sexual subjectivity, there were two limitations worthy of note.			bs6010004_perova	0
+We have done this and have also substantially revised the entire Introduction (see new section starting on p. 3 titled Definition and measurement).	2	1	"Overall, 91% were white/Caucasian, 3% were Asian, 1 was Aboriginal/Pacific Islander, and the remaining participants indicated an ""Other"" sociocultural background."			bs6010004_perova	0
+We have corrected this sentence.	2	1	Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).			bs6010004_perova	0
+We have added to our theoretical explanation for why there would be gender differences (see the section titled Gender Differences in Sexual Behavior, Attitudes, and Sexual Subjectivity, p. 4).	2	1	At Time 1 (T1), young women reported more entitlement than young men.			bs6010004_perova	0
+We have substantially revised all sections of this paper to improve the flow and removed some headings.	2	1	How do the authors make sense of these result, as participants are one year older at Time 2?			bs6010004_perova	0
+We have revised this section (The Current Study, p.5). We have also added hypotheses (see p. 5-6)	2	1	Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.			bs6010004_perova	0
+We have moved this content on group difference to the first section of the Results section.	2	1	Age was positively associated with T1 entitlement, and experience with a wider range of partnered sexual behaviors was concurrently associated with more entitlement and efficacy and was also associated with increased entitlement to partner pleasure and increased self-efficacy in achieving sexual pleasure at T2 relative to T1.			bs6010004_perova	0
+Given our focus on only two element of sexual subjectivity, this was no longer an issue because items were the same on measures for males and females.	2	1	The authors did a nice job of addressing the concerns I pointed out in my previous review.			bs6010004_perova	0
+"We have added this information on p. 7 in the Procedure section. It reads: Approval from the university Human Research Ethics Committee was obtained prior to data collection. At Time 1 (T1), participants were approached at a university campus in Australia in the week before classes commenced (i.e., during orientation week) and asked to participate in a study ""About You and Your Relationships."" The front cover of the survey described the questions as focused on personal sexual and romantic experiences, and stressed the confidential nature of the survey."	2	1	On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.			bs6010004_perova	0
+Reducing the number of elements of sexual subjectivity has reduced the number of tests performed.	2	1	There is also the possibility that individuals more interested in sex and relationships chose to participate.			bs6010004_perova	0
+We have added hypotheses to the section titled The Current Study on p. 5-6.	2	1	On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.			bs6010004_perova	0
+We have now used multiple imputation to maintain all 295 participants in all analyses.	2	1	At Time 1 (T1), young women reported more entitlement than young men.			bs6010004_perova	0
+We are sorry that we did not keep a tally of the number of students approached. We have added to the Limitations section of the Discussion about this (see p. 12). It reads, First, the participants in the study were predominantly middle class, Caucasian university students, residing in one region of Australia. There is also the possibility that individuals more interested in sex and relationships chose to participate. Therefore, the results of the current study may have some limited generalizability.	2	1	The abstract now reads: Abstract: Many scholars have called for an increased focus on positive aspects of sexual health and sexuality.			bs6010004_perova	0
+We agree that this is very important. We have added the Introduction to expand on these points. Unfortunately we could not conduct these suggested analyses. We had very little change in sexual behavior given the way we measured it. Also, we did not have confidence in estimation of sexual behavior for those who did not participate at Wave 2, which could have increased the change. Thus, we have maintained our original analyses, but make this comment in the Discussion (see p. 12-13): Second, probably because the majority of the participants had already experienced coitus prior to the first data collection and because we measured types of sexual behavior and not frequency or some other aspect of behavior, there was high stability in sexual behavior over the one year of this study. Recent statistics show that the majority of Australian year 10-12 students (approximately 15-17 years) have engaged in some form of sexual behavior [12]. Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with change in sexual behavior, future studies may need to begin with a younger population. Recently, Hensel et al. [31] found that, in their adolescent participants, sexual self-concept had a bidirectional relationship with sexual experience, and the same may occur for sexual subjectivity. Future research could examine the possibility of bidirectional relationships between sexual subjectivity and sexual behavior. In addition, no previous study has examined the timing of pubertal development and sexual subjectivity. Evidence suggests that adolescents that mature earlier, compared to their peers, form romantic relationships earlier and experience sexual behaviors earlier [11,55]. In the current and past research, the link between age and sexual subjectivity has been relatively weak [34]. Substituting timing of pubertal status for age may provide more evidence regarding individual characteristics that influence differences in sexual subjectivity and its development over time.	2	1	I think it would be informative to test whether associations with age or sexual experience group are operating through a general underlying factor of sexual subjectivity versus are unique to specific facets.			bs6010004_perova	0
+We have trimmed the paper to focus on two of the five elements of sexual subjectivity and anticipated that these two elements would show different results. Thus, we have maintained the separate analyses of these two elements. Please see the added Hypotheses on p. 5-6.	2	1	At the very least, the Discussion should consider how generalizable these results are.			bs6010004_perova	0
+We have removed these analyses from the paper in an attempt to reduce the number of analyses and streamline the entire paper.	2	1	A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			bs6010004_perova	0
+Thank you for this suggestion. The reference has been added to the paper.	2	1	Analyses should be re-done using a more sophisticated method for handling missing data, such as multiple imputation or full-information maximum likelihood (Shafer & Graham, 2002, Psychological Methods).			bs6010004_perova	0
+"The “inherent affinity” is more appropriate, and the term is updated in the manuscript. Revision in Page 1 Line 28:“The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2].” Point 2:"	2	1	It may be better to find some literature to combine workplace design and “Biophilic Design”, and then to explore the relationship between them.			buildings12040417_makarova	0
+In-text citation in Page 1 Line 44: “Some scholars summarized and classified the natural design features into biophilic design frameworks to guide design activities [25, 39, 40, 50, 51, 52].” Three references are added in the References List: Ulrich, 1993. Biophilia, biophobia, and natural landscapes. The Biophilia hypothesis. USA: Island Press: Washington, D.C. Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. 6, 2332-2343. Ko et al., 2021. A Window View Quality Assessment Framework. Leukos. 1-26. DOI: 10.1080/15502724.2021.1965889. Ko et al., 2021.	2	1	DOI：10.1016/j.buildenv.2020.107443 Point 5: P3, L92: I think refers to “has helped” given the five decades predating this.			buildings12040417_makarova	0
+There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays. And the issue raised by the authors is that these biophilic design frameworks (design guidelines or standards) are general design frameworks (design guidelines or standards) which can be applied to all building typologies (i.e., residential buildings, workplaces, retails, etc). Further research needs to be conducted to develop the design guideline specific for workplace. The sentence was rewritten to demonstrate this argument. Revision in Page 2 Line 56-61: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guidelines and assessment methods are necessary.” Point 4:	2	1	Reading further to page 5, lines 189-192, the authors begin to suggest to this, but referred to them and others indoor environmental parameters as factors for the workplace.			buildings12040417_makarova	0
+We agree with this comment. Apart from the design evaluation, POE is also one of the mainstream research methods that can effectively diagnosing operation problems. The description and citations were inserted in the updated manuscript. Revision in Page 2 Line 77 ~ 80: “Moreover, from the perspective of building operation, the POE results also provide evaluation and feedback from occupants to the stakeholders and building managers on workplace biophilic design. Since POE is one of the mainstream research methods that can effectively diagnosing operation problems [54, 55, 56].” Citations are added in the Reference list: Graham, L.T., Parkinson, T., Schiavon, S., 2021. Lessons Learned from 20 years of CBE’s Occupant Surveys. Buildings and Cities 2(1):166-184. DOI: 10.5334/bc.76 Kent, M., Parkinson, T., Kim, J., Schiavon, S., 2021. A Data-Driven Analysis of Occupant Workspace Dissatisfaction. Building and Environment 205, 108270 Cheung, T., Schiavon, S., Graham, L.T., Tham, K.W., 2021. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. Building and environment (188). DOI：10.1016/j.buildenv.2020.107443 Point 5:	2	1	By contrast, the introduction of “Biophilic Design” is very substantial.			buildings12040417_makarova	0
+The sentence was rewritten in the updated version. Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].”	2	1	At the same time, the conclusion only summarizes the article, and lacks discussions and explanations for the future research direction.			buildings12040417_makarova	0
+Yes, the existing psychological scales are well-developed, but scales for investigating “workplace biophilic design” had not been developed before. Hence, in this study, we develop a method that focus on evaluating the biophilic design elements in workplace. The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE). The questions in the first and the second subscales are referred to the validated scales--The World Health Organization Quality of Life (WHOQOL) and the nature relatedness (NR). The third scale is focus on evaluation on the biophilic design elements. Hence, the questions in the final section are designed based on the selection of the biophilic design elements/attributes that typically applied in the office design, which are not mentioned in the previous scales.	2	1	Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32.			buildings12040417_makarova	0
+The method of selection of the biophilic design attributes/patterns for workplace is: ŸStep one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks. ŸStep two, we neglect the patterns which are not representative in office environment (please find the detailed explanations in the respond for Comment #8) and specify the selected design patterns to nine biophilic design attributes. The detailed process is shown in Section 2.1 and Figure 1. ŸStep three, verify the selection of the nine biophilic attributes matches the validated eight factors that affect workers’ satisfaction and productivity (please find the detailed explanations in the respond for Comment #11). In terms of the reviewer think that the listed attributes span across different domains and the communal features are not that apparent, we believe it is due to the research perspective of biophilic design is different from the perspectives of building science and traditional POE studies (please find the detailed explanations in the respond for Comment #9).	2	1	Lessons learned from 20 years of CBE’s occupant surveys.			buildings12040417_makarova	0
+The authors gave the reason after the sentence in Page 4, Line 173-174 of the original manuscript “Second, the authors neglect seven design patterns from the 14 Patterns of Biophilic Design which are not representative of the workplace design.” to explain why the seven of the patterns from the 14 patterns of biophilic design were discarded: For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended but not demonstrative in this building typology (workplace). Those are usually applicable in other building typologies, such as hotels or residential. To further explain why these patterns are recommended but are discarded: 1) First, these design patterns (i.e., the seven discarded patterns of the 14 biophilic design patterns) are recommended because they are proofed that benefits health. 2) However, these patterns are not common in most offices. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”. 3) Therefore, we only included those biophilic design patterns that relatively easy to apply in the workplace (e.g., greenery, natural light, artworks), and discard those which are not representative in an office design. And the sentences are revised to further explain the discard of the seven patterns. Please see the revised contents in the updated manuscript below: Revision in Page 5 Line 191-197: “For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended because they are proofed that benefits health. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”.” Point 9:	2	1	e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” 2) The Conclusion is reconstructed and a separate section-- Section 6.			buildings12040417_makarova	0
+We believe that there is no conflict between the different classifications. The same parameters (e.g., daylight, thermal comfort, and air-quality, office layout and building form) can be classified in different classifications (i.e., the traditional POE frameworks and the biophilic design frameworks) by different perspectives. 1) From perspective of building science, building performance, and traditional POE, these parameters (e.g., daylight, thermal comfort, and air-quality) are considered as indoor environment parameters, and office layout and building form are considered a physical and architectural parameters. 2) On the other hand, from the perspective of biophilia and biophilic design, (e.g., factors workers’ satisfaction and productivity), these parameters are re-classified and defined as the factors affecting workplace health. Both classifications validated by previous literatures.	2	1	d) The correlation results support the importance of biophilic design from the user perspectives.			buildings12040417_makarova	0
+We revised the Figure 1. The connections between 24 Biophilic Design Attributes (Column A) and 14 Patterns of Biophilic Design (Column B); The nine biophilic design attributes for the workplace (Column C). First, to simplify the image, we put all the definitions into a new table in the appendix (Appendix A. Definitions of biophilic design attributes and patterns). In case that there are some readers are not familiar with the biophilic design attributes or patterns: Revision in Page 4 Line 164: “These two biophilic frameworks are chosen as research references (definitions of the at-tributes and patterns are in Appendix A).” Appendix A in Page 22: “Appendix A. Definitions of biophilic design attributes and patterns” Second, we fill the correlated boxes with same solid colours to make them more recognizable: Revision in Page 6 Line 207: Point 11:	2	1	The sentence was rewritten in the updated version.			buildings12040417_makarova	0
+Table 2 made a comparison between the selected nine biophilic design patterns for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace. These overlapped factors highlight the nine biophilic design attributes that are critical to the office design. Hence, the validation of the selection of the nine biophilic attributes are proofed by the previous literature. Sentences are added in the paragraph to clarify the demonstration. Revision in Page 6 Line 215-220: “Table 2 made a comparison between the selected nine biophilic design attributes for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. These overlapped factors highlight the nine biophilic design at-tributes that are critical to the office design. The validation of the selection of the nine biophilic attributes are proofed by the previous literature [37].” Point 12:	2	1	Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].” Point 6:			buildings12040417_makarova	0
+1) The reasons why the two offices are selected for investigation: a) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices. Revision in Page 9 Line 277-280: “The two offices have similar features: 1) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices.” 2) The supplemental Information (i.e., temperature, number of employees) are added in the revised version. Revision in Page 9 Table 5: Table 5. The structure is re-constructed in the revised version. The original Section 3.1.	2	1	The sentence was rewritten in the updated version.			buildings12040417_makarova	0
+The means (SD) are only applied when the assumption of normality applies for the datasets. Revision in Page 18-20 Table 9: Table 9.	2	1	Lessons learned from 20 years of CBE’s occupant surveys.			buildings12040417_makarova	0
+The Cronbach’s Alpha with an Alpha>0.6 considered acceptable internal consistency in this study. The statement and the references are added in the revised version. Revision in Page 13 Line 381-383: “The Cronbach’s α coefficient value of the main scale is 0.72, while those of the sub-scales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).” Citations are added in the Reference list: Morgan, P. J., Cleave‐Hogg, D., DeSousa, S., Tarshis, J., 2004. High‐fidelity patient simulation: validation of performance checklists. British Journal of Anesthesia, Volume 92, (3) 388–392. Cronbach, L.J., 1951. Coefficient Alpha and the Internal Structure of tests, Psychometrika. Vol. 6. 3. 297-334.	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration.			buildings12040417_makarova	0
+1) Abbreviated labels referring to the actual questions are added in the Figure 3 (Figure 2 in the revision). But 2) we keep the original percentages (round to one decimal place). We think it is not hard for the readers to understand, and the original percentages (round to one decimal place) is more accurate. Revision in Page 17 Line 431:	2	1	If this was the case, perhaps this could be succinctly mentioned.			buildings12040417_makarova	0
+Thanks for the reminder, the error has been corrected in the revised version. Revision in Page 21 Line 473: “According to Table 10, Pearson correlations indicate that…” Point 17:	2	1	Light Glass walls and clerestories, reflecting colors and materials 2.			buildings12040417_makarova	0
+Spearman’s correlation test was conducted, and the revised contents are shown in the updated version. Revision in Page 17 Line 440: “Second, Spearman's correlation analysis is utilized to examine the correlation between three subscales.” Revision in Page 21 Line 472-483: 4.2. Intercorrelation between the three subscales (GH, NR, BDE) “According to Table 10, Spearman's correlations indicate that works' nature relatedness (NR) was positively correlated with self-evaluated GH (r = .264**, p < .01). This result also confirms the previously obtained results that people who had a higher evaluation in nature relatedness are also had a higher evaluation on their health. When the occupants feel that they have a strong sense of relationship with nature, it is observed that the biophilic environment would have positive impacts on their health. More importantly, significant correlation is also found in between biophilic design evaluation and self-reported health (GH), r=.270**, p < .01, indicating that office biophilic design has positive values on workers’ psychological health.” Table 10. Intercorrelations between responses of three subscales. Correlation is significant at the 0.01 level (2-tailed) Point 18: P18, L448-450: Please check whether the sentence is accurate and correct the table caption numbers; I believe these should be Tables 10 and 11 and 12, not 1, 2 and 3.	2	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration.			buildings12040417_makarova	0
+The above contents have been corrected in the revised version. Point 19: While the conclusions were well structured, I felt the authors could have highlighted more the main takeaway messages from their endeavors, in particularly the relationship between biophilic design and occupant health and wellbeing. This seems to be a core aspect of their work but did really emerge from the final section of their work in the same way it was emphasized in the abstract.	2	1	Author Response Response to Reviewer 3 Comments Point 1:			buildings12040417_makarova	0
+The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 514-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Author Response File: Author Response.docx	2	1	Lessons learned from 20 years of CBE’s occupant surveys.			buildings12040417_makarova	0
+In this revised version, we further explain the lack of scales for biophilic design evaluation. Please find the revised sentences as below: Revision in Page 3 Line 122-128: “Actually, these are standard research methods of investigative POE; nevertheless, there are no existing POE scales that focus on biophilic design in the workplace. According to the research objectives of this study (i.e., evaluate the subjective health impacts of biophilic design in workplace), we need to refer the well-developed scales from other disciplinary (e.g., Environmental Psychology). And finally, a scale that integrated health evaluation and building environment evaluation (i.e., POE) is developed for investigation.” Point 2: In consideration to their response #8, I understood and agreed with why half the seven of the 14 papers were discarded, but it did not provide much insight into why the Patterns of Biophilic Design was still used.	4	1	Later (P10, L299), it says 201 questionnaires were collected, with 161 occupants taking part in the Singaporean office.			buildings12040417_makarova	0
+In the revised version, sentences are added to further explain that there are too few framework available, and these two frameworks are the mainstream biophilic design frameworks that are widely applied in the practical biophilic design projects. Revision in Page 3 Line 122-128: “The 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design are the two mainstream biophilic design which are widely applied in the practical biophilic design projects. Hence, these two frameworks are the most suitable to be selected as the basis of this experiment.”	4	1	Images of nature The image and representation of nature in the built environment—plants, animals, landscapes, water, geological features 10.			buildings12040417_makarova	0
+"1) Further explanations of “Biophilia” are added in the Introduction. Revision in Page 1 Line 27-29: “The term ""Biophilia"" is evolved from human evolution research and is coined to describe humans' inherent love affinity for the living things in the natural world [1,2]. It explained why we prefer nature because it is an instinct deeply rooted in the human brain.” 2) Literatures are added in the revised version to combine “workplace design” and “Biophilic Design”. Revision in Page 2 Line 48-53: “Workplace is one of the typologies that attracts the attentions of researchers. Scholars who research the relationship between the built environment and health found that the environment not merely directly or indirectly affects human health but also affects their work and study performance [59]. Studies proofed that biophilic design benefits workers’ health and productivity in an office environment [60, 61, 62, 63].” Citations are added in the Reference list: Derek, C. C., 2003. Environmental Quality and the Productive Workplace. In C. J. Anumba (Ed.), Innovative Developments in Architecture, Engineering and Construction. Rotterdam: Millpress Science Publishers. Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021."	2	1	Biophilia, biophobia, and natural landscapes.			buildings12040417_makarova	0
+"Observation Results-Biophilic Design Attributes in the Selected Offices is moved to be Section 2.3.2. It is because the 2.3.1 is the selection of office, and the Section 2.3.2 provides observation details of the selected offices. After re-construction, Section 3. Questionnaire Results focus on illustrating the questionnaire results. Hence, all the means (SD) are replaced as medians (IQR). And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version. Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8). According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min. 20 to max. 100). Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR. The median (IQR) value of the POE scale BDE is evaluated as 42.00 (5.00) (range of total value: min. 12 to max. 60). Table 8. Medians, Interquartile Range (IQR), Mean, standard deviation, and α coefficient values of workers’ evaluation based on HWBO, GH, NR, and BDE. Comparison of independent variables (Gender, Age, Educational Levels, Weekly Work Hours, Daily Sedentary Time, Work Desk Locations, Working Years, Office Locations) on self-reported GH, NR, and BDE. Thanks for your suggestion. The figures and pictures have been adjusted in the revised manuscript. The Section 3 Questionnaire Results and Section 4 are rewritten in the revised version: a) the statistical analysis is modified; b) to reduce elaboration in the text. Please kindly refer the following revisions: 1) The original Section 3.1. 2) In the revised version, the formulars in Section 3.3 2. Quantitative Results of Impacts of Biophilic Design for Workplace are removed due to the statistical analysis is modified: The means (SD) are only applied when the assumption of normality applies for the datasets. Revision in Page 18 Table 9: Table 9. *p<.05, The significance level is .05.3) Elaboration for the Figure 2. Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement) is reduced, because the illustrations are obvious in the figure. Revision in Page 15-17 Line 399-433: The analysis of individual items provides more details into the works' responses. As can be seen in percentage responses for individual items (Figure 2 in revision), most of the responses are distributed in the items ""Neutral"" and ""Agree"". The questionnaire results reveal that the employees from the understudy companies hold a relatively positive opinion on wellbeing, nature-relatedness, indoor environmental quality, and biophilic design for their health promotion. According to the arrangement, at the top of this, stacked graph are the evaluation of satisfaction of the work capacities and relationships in the workplace. About 73.2% of respondents agree that the workers of the companies under investigation are satisfied with their work capacity (GH3-Q10) and relationships (GH4-Q11). In the subscale nature relatedness, 62.2% and 61.7% of workers responded (agree/strongly agree) that their actions affect the environment (NR2-Q13), and they take notice of the wildlife in their daily lives (NR3-Q14). Nevertheless, only 47.8% of them selected agreed/strongly agree regarding the statements that their ideal spot for vacation would be a wilderness area (NR1-Q12). Regarding the POE results in the subscale BDE, 63.2% of workers agree/strongly agree that the natural light is an essential biophilic attribute and their offices are bright (BDE2-Q17). Furthermore, 60.7 % of the workers agreed that introducing natural colors in the office benefits workplace health and wellbeing (BDE11-Q26). More than 60 percent (approximately 60.7%) of respondents believe that greenery is a biophilic design that benefits office wellbeing (BDE6-Q21). Their feedback would be valuable for designers to note that application of the biophilic design attributes in the office design can enhance the experiences and evaluations of workers. The quantitative results of the questionnaire survey demonstrate that the workers agree that the biophilic design attributes in the office have positive effects on their subjective wellbeing. Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement)."	2	1	Quantitative Results of Impacts of Biophilic Design for Workplace are removed due to the statistical analysis is modified: The means (SD) are only applied when the assumption of normality applies for the datasets.			buildings12040417_makarova	0
+Thank you for your comments. These considerations are the limitations of this study. 1) these two cases are limited in representing all the workplace biophilic designs. As mentioned in the Conclusion, in the future study, we will include more offices and locations as experiment samples. In the future study, the research scope should be narrow down for intensive investigation.	2	1	The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology.			buildings12040417_makarova	0
+Thank you for your suggestion. The description of the necessity of research is highlight in the revised version. Please kindly refer the following revisions: Revision in Page 2 Line 56-66: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guide-lines and assessment methods are necessary. Additionally, the effectiveness of such design in practical design projects for user wellbeing still requires confirmation. More importantly, building typology-based biophilic design guidelines should be appropriately developed because it would affect the designer's prioritization of design attributes selection in design practice.” Point 6:	2	1	Water features Views of prominent water bodies, fountains, aquaria, constructed wetlands 4.			buildings12040417_makarova	0
+The different genders and different ages are included in the study. The age ranges included 21-25, 26-35, 36-45, 46-60. The detailed description of the demographic information is in Section 3.1. Demographic Information.	2	1	Buildings, 11(12), 627. https://doi.org/10.3390/buildings11120627 65.			buildings12040417_makarova	0
+b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices. The weighting results of this study would be especially applicable to the workplace typology. The 14 Patterns of Biophilic Design has a broader range of usage for all building typologies and is more suitable for general design applications. Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design. These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results). c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration. d) The correlation results support the importance of biophilic design from the user perspectives. There is a significant correlation between office biophilic design and self-reported health of employees (r=.270**, p < .01). 2) The investigative POE studies evaluated the self-reported health (GH), nature-relatedness (NR), and biophilic design in the workplace (BDE). The objective of the study is to evaluate the typical biophilic design attributes in office environment and the correlation between biophilic design and office health. Hence, the research scope is relatively extensive. 1) The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” 2) The Conclusion is reconstructed and a separate section-- Section 6. Limitation and Future Studies is added in the revised version: Revision in Page 23 Line 542-551: “6. Limitation and Future Studies There are limitations in this study, first, these two cases are limited in representing all the workplace biophilic designs due to the sample size limitations. Further studies could include more offices and locations as experiment samples. In the future study, the research scope should be narrow down for intensive investigation.” Point 8:	2	1	The weighting results of this study would be especially applicable to the workplace typology.			buildings12040417_makarova	0
+The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology. And the questionnaire can be applied in future biophilic design research for investigation. The applications of the results are highlighted in Section 1.4. Objectives and in Section 5 Conclusion: In Section 1.4. Objectives, the three practical applications of the results are mentioned: first, a POE questionnaire for assessing the biophilic design for workplace accounting for health and wellbeing. Second, the study will provide a new biophilic design guidelines for workplaces, which can effectively assist researchers and designers to improve office biophilic design practices and decision-making on design attributes selection. The Conclusion is rewritten to highlight the contribution and implementation of the results: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. … “c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration.” … “e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Point 9: It is better to supplement the temperature, humidity, and other parameters of the selected office in the part of the experiment, so as to facilitate readers' reference rather than just giving the location.	2	1	Survey measures Category GH NR BDE Median (IQR) Sig.			buildings12040417_makarova	0
+Basic information of the understudy offices. The supplemental Information (i.e., office temperature, number of employees) are added in the revised version. Revision in Table 5: Table 5.	2	1	Biophilic design is known to elicit mental and physical health benefits, as stated by the authors on page 1, lines 29-32.			buildings12040417_makarova	0
+We agree with the comment. A quantitative study for indoor workplace biophilic design to improve health and productivity performance. Journal of Cleaner Production. 324, 129168. Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018. Physiological and cognitive performance of exposure to biophilic indoor environment. Build. Environ. 132, 255–262. Yin, J., Arfaei, N., MacNaughton, P., Catalano, P.J., Allen, J.G., Spengler, J.D., 2019. Effects of biophilic interventions in office on stress reaction and cognitive function: a randomized crossover study in virtual reality. Indoor Air 29, 1028–1039. https:// doi.org/10.1111/ina.12593. Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020. Effects of biophilic indoor environment on stress and anxiety recovery: a between-subjects experiment in virtual reality. Environ. Int. 136, 105427. https://doi.org/10.1016/j. envint.2019.105427. Previous studies proofed that biophilic design in workplaces benefits both psychological and physiological health [60, 61, 62, 63]. This study is a questionnaire survey that focus on investigating the subjective evaluation on workplace biophilic design and of workers. Physiological measurements will be included in the future study to investigate the impacts of the psychological factors on physiological comfort. References: Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021.	2	1	In the subscale nature relatedness, 62.2% and 61.7% of workers responded (agree/strongly agree) that their actions affect the environment (NR2-Q13), and they take notice of the wildlife in their daily lives (NR3-Q14).			buildings12040417_makarova	0
+Thank you for your comment. Figure 1 is updated in this version.	4	1	Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020.			buildings12040417_makarova	0
+All the photos in Table 6 are taken by the designers. And we added an annotation under Table 6.	4	1	2) The investigative POE studies evaluated the self-reported health (GH), nature-relatedness (NR), and biophilic design in the workplace (BDE).			buildings12040417_makarova	0
+"Thank you for your suggestions. The references are added in the manuscript. In-text citations in Page 1 Line 27-28: “The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2,64].” In-text citations in Page 1 Line 40: “…the modern built environment' as 'Biophilic Design' [24,25,65].” In-text citations in Page 2 Line 69-70: “…occupant satisfaction, health, and wellbeing after occupancy of buildings [27, 28, 67].” Citations are added in the Reference list: Parsaee, M., Demers, M. H. C., Potvin, A., Hébert, M., Lalonde, J.F., Window View Access in Architecture: Spatial Visualization and Probability Evaluations Based on Human Vision Fields and Biophilia. 2021. Buildings, 11(12), 627. https://doi.org/10.3390/buildings11120627 Mollazadeh, M., Zhu, YM., Application of Virtual Environments for Biophilic Design: A Critical Review. 2021. Buildings, 11(4), 148; https://doi.org/10.3390/buildings11040148 Gillis, K., Gatersleben, B., 2015. A Review of Psychological Literature on the Health and Wellbeing Benefits of Biophilic Design. Buildings. 5(3), 948-963. https://doi.org/10.3390/buildings5030948 Candido, C., Chakraborty, P., Tjondronegoro, D., 2019. The Rise of Office Design in High-Performance, Open-Plan Environments. 9(4), 100. https://doi.org/10.3390/buildings9040100 Author Response File: Author Response.docx"	4	1	Some of the results in the illustrations are obvious and can be reduced to less elaboration.			buildings12040417_makarova	0
+"The “inherent affinity” is more appropriate, and the term is updated in the manuscript. Revision in Page 1 Line 28:“The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2].” Point 2:"	2	1	The age ranges included 21-25, 26-35, 36-45, 46-60.			buildings12040417_perova	0
+Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. Ulrich, 1993. Biophilia, biophobia, and natural landscapes. Ko et al., 2020. A window view quality assessment framework. LEUKOS. In-text citation in Page 1 Line 44: “Some scholars summarized and classified the natural design features into biophilic design frameworks to guide design activities [25, 39, 40, 50, 51, 52].” Three references are added in the References List: Ulrich, 1993. Biophilia, biophobia, and natural landscapes. The Biophilia hypothesis. USA: Island Press: Washington, D.C. Bjørn et al., 2009. Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being? International Journal of Environmental Research and Public Health. 6, 2332-2343. Ko et al., 2021. A Window View Quality Assessment Framework. Leukos. 1-26. DOI: 10.1080/15502724.2021.1965889. Ko et al., 2021.	2	1	In the future study, the research scope should be narrow down for intensive investigation.			buildings12040417_perova	0
+There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays. And the issue raised by the authors is that these biophilic design frameworks (design guidelines or standards) are general design frameworks (design guidelines or standards) which can be applied to all building typologies (i.e., residential buildings, workplaces, retails, etc). Further research needs to be conducted to develop the design guideline specific for workplace. The sentence was rewritten to demonstrate this argument. Revision in Page 2 Line 56-61: “Although the importance of biophilic design seems to be well-acknowledged, and some international or regional green building and healthy building standards incorporate biophilic design elements into the rating system, such as WELL building standard version 2 and Singapore Green Mark [53]. However, further research on developing building typology-based biophilic design guidelines and assessment methods are necessary.” Point 4:	2	1	Introduction #1: The definition of biophilia is described as an “inherent love” toward nature.			buildings12040417_perova	0
+Lessons learned from 20 years of CBE’s occupant surveys. A data-driven analysis of occupant workspace dissatisfaction. Occupant satisfaction with the indoor environment in seven commercial buildings in Singapore. We agree with this comment. Apart from the design evaluation, POE is also one of the mainstream research methods that can effectively diagnosing operation problems. The description and citations were inserted in the updated manuscript. Revision in Page 2 Line 77 ~ 80: “Moreover, from the perspective of building operation, the POE results also provide evaluation and feedback from occupants to the stakeholders and building managers on workplace biophilic design. Since POE is one of the mainstream research methods that can effectively diagnosing operation problems [54, 55, 56].” Citations are added in the Reference list: Graham, L.T., Parkinson, T., Schiavon, S., 2021. Buildings and Cities 2(1):166-184. DOI: 10.5334/bc.76 Kent, M., Parkinson, T., Kim, J., Schiavon, S., 2021. Building and Environment 205, 108270 Cheung, T., Schiavon, S., Graham, L.T., Tham, K.W., 2021. Building and environment (188). DOI：10.1016/j.buildenv.2020.107443 Point 5:	2	1	Those are usually applicable in other building typologies, such as hotels or residential.			buildings12040417_perova	0
+The sentence was rewritten in the updated version. Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].” Point 6:	2	1	Transitional spaces hallways, thresholds, doorways, gateways, and areas that link the indoors and outdoors especially porches, patios, courtyards, colonnades 23.			buildings12040417_perova	0
+Yes, the existing psychological scales are well-developed, but scales for investigating “workplace biophilic design” had not been developed before. Hence, in this study, we develop a method that focus on evaluating the biophilic design elements in workplace. The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE). The questions in the first and the second subscales are referred to the validated scales--The World Health Organization Quality of Life (WHOQOL) and the nature relatedness (NR). The third scale is focus on evaluation on the biophilic design elements. Hence, the questions in the final section are designed based on the selection of the biophilic design elements/attributes that typically applied in the office design, which are not mentioned in the previous scales.	2	1	It is best to supplement and describe the necessity of research.			buildings12040417_perova	0
+The method of selection of the biophilic design attributes/patterns for workplace is: ŸStep one, find out the correlated biophilic design characteristics from the two mainstream biophilic design frameworks. ŸStep two, we neglect the patterns which are not representative in office environment (please find the detailed explanations in the respond for Comment #8) and specify the selected design patterns to nine biophilic design attributes. The detailed process is shown in Section 2.1 and Figure 1. ŸStep three, verify the selection of the nine biophilic attributes matches the validated eight factors that affect workers’ satisfaction and productivity (please find the detailed explanations in the respond for Comment #11). In terms of the reviewer think that the listed attributes span across different domains and the communal features are not that apparent, we believe it is due to the research perspective of biophilic design is different from the perspectives of building science and traditional POE studies (please find the detailed explanations in the respond for Comment #9).	2	1	According to the arrangement, at the top of this, stacked graph are the evaluation of satisfaction of the work capacities and relationships in the workplace.			buildings12040417_perova	0
+The authors gave the reason after the sentence in Page 4, Line 173-174 of the original manuscript “Second, the authors neglect seven design patterns from the 14 Patterns of Biophilic Design which are not representative of the workplace design.” to explain why the seven of the patterns from the 14 patterns of biophilic design were discarded: For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended but not demonstrative in this building typology (workplace). Those are usually applicable in other building typologies, such as hotels or residential. To further explain why these patterns are recommended but are discarded: 1) First, these design patterns (i.e., the seven discarded patterns of the 14 biophilic design patterns) are recommended because they are proofed that benefits health. 2) However, these patterns are not common in most offices. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”. 3) Therefore, we only included those biophilic design patterns that relatively easy to apply in the workplace (e.g., greenery, natural light, artworks), and discard those which are not representative in an office design. And the sentences are revised to further explain the discard of the seven patterns. Please see the revised contents in the updated manuscript below: Revision in Page 5 Line 191-197: “For instance, the patterns “Presence of Water”, “Prospect”, “Refuge”, “Mystery”, and “Risk” (i.e., itemized patterns 5, 11, 12, 13, and 14 in Column B) are recommended because they are proofed that benefits health. For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”.” Point 9:	2	1	Response 2: All the photos in Table 6 are taken by the designers.			buildings12040417_perova	0
+We believe that there is no conflict between the different classifications. The same parameters (e.g., daylight, thermal comfort, and air-quality, office layout and building form) can be classified in different classifications (i.e., the traditional POE frameworks and the biophilic design frameworks) by different perspectives. 1) From perspective of building science, building performance, and traditional POE, these parameters (e.g., daylight, thermal comfort, and air-quality) are considered as indoor environment parameters, and office layout and building form are considered a physical and architectural parameters. 2) On the other hand, from the perspective of biophilia and biophilic design, (e.g., factors workers’ satisfaction and productivity), these parameters are re-classified and defined as the factors affecting workplace health. Both classifications validated by previous literatures.	2	1	9. material Connection with nature Material and elements from nature that, through minimal processing, reflect the local ecology or geology to create a distinct sense of place.			buildings12040417_perova	0
+We revised the Figure 1. The connections between 24 Biophilic Design Attributes (Column A) and 14 Patterns of Biophilic Design (Column B); The nine biophilic design attributes for the workplace (Column C). First, to simplify the image, we put all the definitions into a new table in the appendix (Appendix A. Definitions of biophilic design attributes and patterns). In case that there are some readers are not familiar with the biophilic design attributes or patterns: Revision in Page 4 Line 164: “These two biophilic frameworks are chosen as research references (definitions of the at-tributes and patterns are in Appendix A).” Appendix A in Page 22: “Appendix A. Definitions of biophilic design attributes and patterns” Second, we fill the correlated boxes with same solid colours to make them more recognizable: Revision in Page 6 Line 207: Point 11:	2	1	Scholars who research the relationship between the built environment and health found that the environment not merely directly or indirectly affects human health but also affects their work and study performance [59].			buildings12040417_perova	0
+Table 2 made a comparison between the selected nine biophilic design patterns for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace. These overlapped factors highlight the nine biophilic design attributes that are critical to the office design. Hence, the validation of the selection of the nine biophilic attributes are proofed by the previous literature. Sentences are added in the paragraph to clarify the demonstration. Revision in Page 6 Line 215-220: “Table 2 made a comparison between the selected nine biophilic design attributes for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity. These overlapped factors highlight the nine biophilic design at-tributes that are critical to the office design. The validation of the selection of the nine biophilic attributes are proofed by the previous literature [37].” Point 12:	2	1	Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design.			buildings12040417_perova	0
+1) The reasons why the two offices are selected for investigation: a) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices. Revision in Page 9 Line 277-280: “The two offices have similar features: 1) the urban contexts are similar: both the cities (Singapore and Shenzhen) are typical compact, high density Asian mega-cities; b) both the offices are open-plan offices.” 2) The supplemental Information (i.e., temperature, number of employees) are added in the revised version. Revision in Page 9 Table 5: Table 5. Basic information of the understudy offices. The structure is re-constructed in the revised version. The original Section 3.1.	2	1	b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices.			buildings12040417_perova	0
+We agree with the comment. The means (SD) are only applied when the assumption of normality applies for the datasets. According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min. 20 to max. 100). Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR. The median (IQR) value of the POE scale BDE is evaluated as 42.00 (5.00) (range of total value: min. 12 to max. 60). Table 8. Medians, Interquartile Range (IQR), Mean, standard deviation, and α coefficient values of workers’ evaluation based on HWBO, GH, NR, and BDE. Revision in Page 18-20 Table 9: Table 9. Comparison of independent variables (Gender, Age, Educational Levels, Weekly Work Hours, Daily Sedentary Time, Work Desk Locations, Working Years, Office Locations) on self-reported GH, NR, and BDE.	2	1	Comparison of independent variables (Gender, Age, Educational Levels, Weekly Work Hours, Daily Sedentary Time, Work Desk Locations, Working Years, Office Locations) on self-reported GH, NR, and BDE.			buildings12040417_perova	0
+Thank you for your suggestion. The Cronbach’s Alpha with an Alpha>0.6 considered acceptable internal consistency in this study. The statement and the references are added in the revised version. Revision in Page 13 Line 381-383: “The Cronbach’s α coefficient value of the main scale is 0.72, while those of the sub-scales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).” Citations are added in the Reference list: Morgan, P. J., Cleave‐Hogg, D., DeSousa, S., Tarshis, J., 2004. High‐fidelity patient simulation: validation of performance checklists. British Journal of Anesthesia, Volume 92, (3) 388–392. Cronbach, L.J., 1951. Coefficient Alpha and the Internal Structure of tests, Psychometrika. Vol. 297-334.	2	1	And the questionnaire can be applied in future biophilic design research for investigation.			buildings12040417_perova	0
+1) Abbreviated labels referring to the actual questions are added in the Figure 3 (Figure 2 in the revision). But 2) we keep the original percentages (round to one decimal place). We think it is not hard for the readers to understand, and the original percentages (round to one decimal place) is more accurate. Revision in Page 17 Line 431: Point 16:	2	1	Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR.			buildings12040417_perova	0
+Thanks for the reminder, the error has been corrected in the revised version. Revision in Page 21 Line 473: “According to Table 10, Pearson correlations indicate that…” Point 17:	2	1	Revision in Page 3 Line 122-128: “The 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design are the two mainstream biophilic design which are widely applied in the practical biophilic design projects.			buildings12040417_perova	0
+Spearman’s correlation test was conducted, and the revised contents are shown in the updated version. Revision in Page 17 Line 440: “Second, Spearman's correlation analysis is utilized to examine the correlation between three subscales.” Revision in Page 21 Line 472-483: 4.2. Intercorrelation between the three subscales (GH, NR, BDE) “According to Table 10, Spearman's correlations indicate that works' nature relatedness (NR) was positively correlated with self-evaluated GH (r = .264**, p < .01). This result also confirms the previously obtained results that people who had a higher evaluation in nature relatedness are also had a higher evaluation on their health. When the occupants feel that they have a strong sense of relationship with nature, it is observed that the biophilic environment would have positive impacts on their health. More importantly, significant correlation is also found in between biophilic design evaluation and self-reported health (GH), r=.270**, p < .01, indicating that office biophilic design has positive values on workers’ psychological health.” Table 10. Intercorrelations between responses of three subscales. Correlation is significant at the 0.01 level (2-tailed) Point 18:	2	1	"Revision in Page 11 Line 339: “(Photos: by the authors)” Point 3: Please bring strong relevance to the scope of journal ""Buildings"" by investigating most recent literature."			buildings12040417_perova	0
+Thank you for your comment. The above contents have been corrected in the revised version.	2	1	Figure 1 provides some insights into this, but these listed attributes span across different domains and the communal features are not that apparent.			buildings12040417_perova	0
+The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 514-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing. The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology. And the questionnaire can be applied in future biophilic design research for investigation. b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices. The weighting results of this study would be especially applicable to the workplace typology. The 14 Patterns of Biophilic Design has a broader range of usage for all building typologies and is more suitable for general design applications. Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design. These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results). c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration. d) The correlation results support the importance of biophilic design from the user perspectives. There is a significant correlation between office biophilic design and self-reported health of employees (r=.270**, p < .01). e) The study results contribute to provide designers with evidence-based design at-tributes for workplace design (i.e., the nine selected workplace biophilic design attributes).” Author Response File: Author Response.docx	2	1	In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues.			buildings12040417_perova	0
+"1) Further explanations of “Biophilia” are added in the Introduction. Revision in Page 1 Line 27-29: “The term ""Biophilia"" is evolved from human evolution research and is coined to describe humans' inherent love affinity for the living things in the natural world [1,2]. It explained why we prefer nature because it is an instinct deeply rooted in the human brain.”"	2	1	There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace.			buildings12040417_perova	0
+2) Literatures are added in the revised version to combine “workplace design” and “Biophilic Design”. Revision in Page 2 Line 48-53: “Workplace is one of the typologies that attracts the attentions of researchers. Scholars who research the relationship between the built environment and health found that the environment not merely directly or indirectly affects human health but also affects their work and study performance [59]. Studies proofed that biophilic design benefits workers’ health and productivity in an office environment [60, 61, 62, 63].” Citations are added in the Reference list:	2	1	Reading further to page 3, I believe these are the 24 biophilic design attributes [ref.			buildings12040417_perova	0
+Derek, C. C., 2003. Environmental Quality and the Productive Workplace. In C. J. Anumba (Ed.), Innovative Developments in Architecture, Engineering and Construction. Rotterdam: Millpress Science Publishers. Lei, Q.H., Yuan, C., Lau, S.S.Y., 2021. A quantitative study for indoor workplace biophilic design to improve health and productivity performance. Journal of Cleaner Production. 324, 129168. Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018. Physiological and cognitive performance of exposure to biophilic indoor environment. Build. Environ. 132, 255–262. Yin, J., Arfaei, N., MacNaughton, P., Catalano, P.J., Allen, J.G., Spengler, J.D., 2019. Effects of biophilic interventions in office on stress reaction and cognitive function: a randomized crossover study in virtual reality. Indoor Air 29, 1028–1039. https:// doi.org/10.1111/ina.12593. Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020. Effects of biophilic indoor environment on stress and anxiety recovery: a between-subjects experiment in virtual reality. Int. 136, 105427. https://doi.org/10.1016/j. envint.2019.105427. 61.7% of workers responded (agree/strongly agree) that their actions affect the environment (NR2-Q13), and they take notice of the wildlife in their daily lives (NR3-Q14). 63.2% of workers agree/strongly agree that the natural light is an essential biophilic attribute and their offices are bright (BDE2-Q17). 60.7 % of the workers agreed that introducing natural colors in the office benefits workplace health and wellbeing (BDE11-Q26). Citations are added in the Reference list: 59.	2	1	And the questionnaire can be applied in future biophilic design research for investigation.			buildings12040417_perova	0
+Observation Results-Biophilic Design Attributes in the Selected Offices is moved to be Section 2.3.2. It is because the 2.3.1 is the selection of office, and the Section 2.3.2 provides observation details of the selected offices. After re-construction, Section 3. Questionnaire Results focus on illustrating the questionnaire results. Hence, all the means (SD) are replaced as medians (IQR). And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version. Thanks for your suggestion. The figures and pictures have been adjusted in the revised manuscript. The Section 3 Questionnaire Results and Section 4 are rewritten in the revised version: a) the statistical analysis is modified; b) to reduce elaboration in the text. Please kindly refer the following revisions: 1) The original Section 3.1. 2) In the revised version, the formulars in Section 3.3 2. Quantitative Results of Impacts of Biophilic Design for Workplace are removed due to the statistical analysis is modified: The means (SD) are only applied when the assumption of normality applies for the datasets.	2	1	Yin, J., Yuan, J., Arfaei, N., Catalano, P.J., Allen, G.J., Spengler, J.D., 2020.			buildings12040417_perova	0
+Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).	2	1	Response 16: Thanks for the reminder, the error has been corrected in the revised version.			buildings12040417_perova	0
+Revision in Page 18 Table 9: Table 9.	2	1	According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min.			buildings12040417_perova	0
+Revision in Page 15-17 Line 399-433: The analysis of individual items provides more details into the works' responses.	2	1	The two offices present interesting case studies, but it wasn’t made clear why they were selected and how they served the research aim (comment #12).			buildings12040417_perova	0
+Figure 1 is updated in this version.	4	1	A quantitative study for indoor workplace biophilic design to improve health and productivity performance.			buildings12040417_perova	0
+All the photos in Table 6 are taken by the designers. And we added an annotation under Table 6.	4	1	An image showing the indoor conditions and outdoor façade for each might be beneficial.			buildings12040417_perova	0
+Thank you for your suggestions. The references are added in the manuscript.	4	1	This led me to believe that this office was much larger than the building studied in China.			buildings12040417_perova	0
+Subtotal resection is usually defined as a resection of 95-90%, with <90% defined as partial. We addended the submission in line 93-95 as well as table 4 as recommended.	2	1	That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or).			cancers14040966_makarova	0
+We are grateful to the reviewer for catching these mistakes. Both volume reduction and pathology associated numbers are formatting errors and we addended the table to correct these mistakes.	2	1	As we all know surgery of chordomas, especially of recurrent chordomas, is challenging and the authors should be congratulated for their thoroughly surgical planning and approaches, using elaborate pre- and intraoperative imaging, which is well described throughout the manuscript.			cancers14040966_makarova	0
+We agree with the reviewer that useful tools and techniques are available in different modalities and certainly surgeons dealing with various tumors utilize these as needed. We are fortunate to have access to this multi-modality image guided operating suite, and here we report on its use in specific rare tumors, chordomas and chondrosarcomas. We agree that the use of the PET-CT has no use in these cases, and we mentioned it only as part of the general introduction of the AMIGO suite. We addended the manuscript (line 260-264) as recommended	2	1	Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions.			cancers14040966_makarova	0
+We agree with the reviewer that it is the acquisition of multimodality (CT and MRI), with registration of preoperative imaging then intraoperative co-registration. We shared these steps in the “Operative and perioperative management” section (line 60).	2	1	First, when one reads it initially it would appear that there are many different modalities available for intraoperative imaging.			cancers14040966_makarova	0
+As acknowledged in the limitation section and noted by the reviewer, the rarity of these tumors and the small number of patients in most series, including ours, limits analysis. Hence, we did not aim to generalize our outcome. Our focus was on how the use of these modern tools can facilitate maximum safe tumor removal which is considered a pivotal factor in the management of chordoma and chondrosarcoma.	2	1	The results, while admirable, cannot easily be compared to other series, as the numbers are small and each case has its own complex series of characteristics that make it unique.			cancers14040966_makarova	0
+Subtotal resection is usually defined as a resection of 95-90%, with <90% defined as partial. We addended the submission in line 93-95 as well as table 4 as recommended.	2	1	Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions.			cancers14040966_perova	0
+We are grateful to the reviewer for catching these mistakes. Both volume reduction and pathology associated numbers are formatting errors and we addended the table to correct these mistakes.	2	1	With these caveats clarified in a modified version, the paper is valuable and could be published.			cancers14040966_perova	0
+We agree with the reviewer that it is the acquisition of multimodality (CT and MRI), with registration of preoperative imaging then intraoperative co-registration. We shared these steps in the “Operative and perioperative management” section (line 60).	2	1	The authors appear to really only use CT and MRI intraoperatively, with the other modalities imported preoperatively into the Brain Lab navigational system.			cancers14040966_perova	0
+As acknowledged in the limitation section and noted by the reviewer, the rarity of these tumors and the small number of patients in most series, including ours, limits analysis. Hence, we did not aim to generalize our outcome. Our focus was on how the use of these modern tools can facilitate maximum safe tumor removal which is considered a pivotal factor in the management of chordoma and chondrosarcoma.	2	1	However, chordoma is a rare disease and reports on large patient series are rare.			cancers14040966_perova	0
+As suggested, we analyzed the distribution of IRS (immunoreactivity score) across the cohort. In Figure 1 it can be clearly seen that there are two groups forming here, those with 0-4 and those with 6-12 IRS. In addition, we calculated the survival probability depending on the IRS (Figure 2). Patients with an IRS of 6 or higher have a similar bad prognosis compared to patients with an IRS below 5. A better survival correlates with lower MACC1 IRS. Patients with an IRS of 0 have the best prognosis. Based on these two analyses, the decision for cut-off value of 5 was made. Additionally, ROC curve analysis showed, that the IRS value of 5 has the highest Youden Index and is therefore the most suitable cut-off value. 0.408) (Figure 3 and Table 1). Looking at MACC1 expression in cell lines and tissue samples, the variability looks similar. Nevertheless, it must be considered that the investigation of the homogeneous cell lines is a purely quantitative analysis of the expression by Western plot and RT-PCR. The analysis of the tissue samples by immunoreactivity score analyses both the strength of expression and the inhomogeneity of expression. Figure 1: Distribution of the IRS across the cohort Figure 2: Overall Survival depending on the IRS Figure 3: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Table 1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	2	1	The results are interesting and may have clinical relevance.			cancers14071773_makarova	0
+We thank the reviewer for raising this point. However, to respond to this question appropriately, we currently can provide only the results from proliferation assays for one cell line model (FLO1-EV vs FLO1-MACC1) over the time period of 72 h. This assay was with no statistically significant difference in cell proliferation between FLO1-EV vs FLO1-MACC1 (see Figure 4). Figure 4: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay. Unfortunately, the short revision period of 10 days limits the generation of additional data sets for proliferation in further tumor cell lines.	2	1	"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e."			cancers14071773_makarova	0
+We thank the reviewer for this question. The concept of MACC1 inhibition was originally developed by our group for the treatment of MACC1 positive colorectal cancers and was recently published (Kobelt et al. Oncogene 2021). In this study we transferred our experience with MACC1 inhibition by selumetinib on AGE/S. Therefore, we chose the same doses for in vitro and in vivo experiments, and which was also used by other authors (Huynh et al. Mol Cancer Therapy. 2007). Further, we knew that the selected selumetinib concentrations of 10 µM has no cytotoxic effect in vitro and showed no reduction of migration in the MACC1 negative clones (FLO1-EV and OEshMACC1). Regarding the different dosages in the context of xenografted mice treatment in vivo and the clinically used dosages, the FDA recommends multiplying the dosage by a factor of 0.08 when transferring from mouse to human (Niar et al. Journal of basic and clinical pharmacy 2016). According to this calculation, the dosage of 50 mg selumetinib /kg mouse used would equal a dosage of 4 mg/kg in humans. The clinical dose of 75mg twice daily corresponds to a dose of 2x1mg per kg assuming a patient weight of 75kg. Using the aforementioned calculation, this dose equals then 12.5 mg/kg for treatment of mice. In our study, we refer again to the work of Huynh et al. Here, in which a dosage of 12 mg/kg selumetinib showed only a low level of effectiveness. At 100 mg/kg, a very good effectiveness was seen, however combined with an increased toxicity. Thus, we used in our study the dosage of 50 mg/kg as an effective dosage with low toxicity for the mice. Overall, we are considering here a pre-clinical model. The possibility of MEK1 inhibition to inhibit MACC1-induced effects in AGE/S was examined. Further, extended in vivo studies might then reveal the ideal dosage in vivo, being thereafter translated to use in humans. The use of trametinib is an interesting suggestion and might well be adapted to its use in forthcoming studies References: Kobelt, D., Perez-Hernandez, D., Fleuter, C., Dahlmann, M., Zincke, F., Smith, J., ... & Stein, U. (2021). The newly identified MEK1 tyrosine phosphorylation target MACC1 is druggable by approved MEK1 inhibitors to restrict colorectal cancer metastasis. Oncogene, 40(34), 5286-5301. Huynh H, Soo KC, Chow PK, Tran E. Targeted inhibition of the extracellular signal-regulated kinase kinase pathway with AZD6244 (ARRY-142886) in the treatment of hepatocellular carcinoma. Mol Cancer Ther. 2007 Jan;6(1):138-46.. Nair, Anroop B.; Jacob, Shery. (2016). A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy, 7. Jg., Nr. 2, S. 27.	2	1	If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			cancers14071773_makarova	0
+We thank the reviewer for this comment. The IRS cut-off value 5 reaches a sensitivity of 76.6% and a specificity of 64.2% (Youden Index. We include the arguments into the methods section page 3: To define the cut-off value a ROC curve analysis was performed. 0.408) and was chose as cut-off value (see supplement Figure S1 and Table S1). Additionally, we added the ROC curve and the table with sensitivity, specificity and Youden Index into a supplement section as figure S1 and table S1 Figure S1: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Positive if Greater than or Equal to Sensitivity 1- Specificity Specificity Youden Index -1 1 1 0 0 1 0,869 0,500 0,500 0,369 3 0,864 0,483 0,517 0,381 5 0,766 0,358 0,642 0,408 7 0,729 0,342 0,658 0,387 8,5 0,421 0,217 0,783 0,204 10,5 0,411 0,200 0,800 0,211 13 0 0 1 0 Table S1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	4	1	"should be after ""Expression was correlated with survival and morphological characteristics."""			cancers14071773_makarova	0
+thank you for this comment: we added the proliferation analyses by MTT into the method section page 5: Additionally, proliferation for FLO-1/EV and FLO-1/MACC1 was analyzed over 72 h by MTT. 4x103 cells were plated into 96-well-plates and were allowed to accommodate for 24 h. after 48 and 72 h formazan crystals were dissolved in 150 μl of DMSO and the absorption was measured at 560 nm in the absorbance reader (Tecan infinite 200 PRO). Each cell proliferation experiment was performed in triplicates. We added the results of the MTT analyses into the results section page 11: The additional analysis of the proliferation over 72 h showed no significant differences between FLO-1/EV (100% ± 4.886) and FLO-1/MACC1 (112.3% ± 12.24; p=0.113) (see supplement Figure S3). We added the MTT plot as figure S2 into the supplement section Figure S2: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay.	4	1	[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			cancers14071773_makarova	0
+We apologize for the inconvenience. Figure 4 can be found on page 14 in the submitted version. We inserted Figure 4 in the revised manuscript again to solve the technical issue.	2	1	If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			cancers14071773_makarova	0
+We address this question in the results part MACC1 expression: “ Due to the procedure of cutting and staining, some samples could not be used for the evaluation. Samples with insufficiently representative tumor tissue were also excluded from the evaluation. Cores with representative tumor material and evaluable staining were available in 266 of 360 samples (73.9%)…”. To avoid misunderstandings, we corrected the abstract to: 266 Samples of 360 AGE/S patients were analyzed for MACC1 expression.	2	1	"""105"" instead of ""10^5"" in lines 152 and 170, ""106"" instead of ""10^6"" in line 217, ""X2"" instead of ""X^2"" in line 241, ""108"" instead of ""10^8"" in lines 353, 354, 365, 366, 367, 368, 370, 371)."			cancers14071773_makarova	0
+We thank the reviewer for this correction. This formatting issue was corrected throughout the entire manuscript.	2	1	It would be nice to know the selumetinib sensitivity of the two used cell lines.			cancers14071773_makarova	0
+We thank the reviewer for this valuable hint. As suggested, we included p-values in Figure 3G and 3H. In Figure 3G there is a significant increase in migration of cells without MACC1 expression. This is based on a sufficiently large N, that renders even small differences statistically significant. Further, this increase is rather small and is outperformed by the MACC1-mediated increase in migration and the reduction in the MACC1-mediated migration by selumetinib. The data show, that selumetinib does not reduce migration of the MACC1-negative FLO1 and that MACC1 induces migration in this cell line that can be inhibited by selumetinib.	2	1	[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.			cancers14071773_makarova	0
+We thank the reviewer for this suggestion to improve the visual quality of the manuscript. The legends of 3F and I have been enlarged and the resolution of the figure has been increased.	2	1	Minor Issues: [4] In Figures 3G and 3H, please specify p-values for untreated vs selumetinib in FLO-1/EV and OE33/shMACC1 respectively.			cancers14071773_makarova	0
+We thank the reviewer for this discussion. The empty vector control clone was transduced and selected afterwards. During the selection procedure it is possible that a subclone was generated that expresses slightly more (or less) of any mRNA and protein. It differs from the wildtype by handling (transduction, selection etc) and somewhat by age (cell divisions). Therefore, for all comparisons the empty vector clone was used, as this clone was generated in parallel to the MACC1 clone.	2	1	In the pdf I reviewed, the caption is there, but the figure is not.			cancers14071773_makarova	0
+We thank the reviewer for pointing this out: Survival reports were changed in median survival and the 95% confidence interval were included. Additionally, disease-specific survival results were reported.	2	1	[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values).			cancers14071773_makarova	0
+We thank the reviewer for pointing this out: OR with the confidence interval is now included.	2	1	Please discuss how the applied concentrations are related to the clinically used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.			cancers14071773_makarova	0
+We thank the reviewer for pointing this out: all survival reports were changed in median survival and the 95% confidence interval were included.	2	1	The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50 mg/kg) are quite high.			cancers14071773_makarova	0
+Thank you for this valuable comment. The journal requires an unstructured abstract. As suggested by the reviewer, we have therefore reorganized the abstract and improved the flow of reading: Abstract: Esophageal and Gastric Adenocarcinomas (AGE/S) are characterized by early metastasis and poor survival. MACC1 (Metastasis Associated in Colon Cancer 1) acts in colon cancer as a metastasis inducer, linked to reduced survival. This project illuminates the role and potential of inhibition of MACC1 in AGE/S. Using 266 of 360 TMAs and survival data of AGE/S patients we confirm the value of MACC1 as an independent negative prognostic marker in AGE/S patients. MACC1 gene expression correlated with survival and morphological characteristics. In vitro analysis of lentivirally MACC1 manipulated subclones of FLO-1 and OE33 showed enhanced migration induced by MACC1 in both cell line models, which could be inhibited by the MEK1 inhibitor selumetinib. In vivo, the efficacy of selumetinib on tumor growth and metastasis of MACC1-overexpressing FLO-1 cells xenografted intrasplenically in NOG mice was tested. Mice with high MACC1 expressing cells developed faster and larger distant metastases. Treatment with selumetinib led to a significant reduction of metastasis exclusively in the MACC1 positive xenografts.	2	1	Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?			cancers14071773_makarova	0
+There were no more open questions in the second review.	4	1	[3] Throughout the paper, 10^x wrongly displays as 10x (e.g.			cancers14071773_makarova	0
+As suggested, we analyzed the distribution of IRS (immunoreactivity score) across the cohort. In Figure 1 it can be clearly seen that there are two groups forming here, those with 0-4 and those with 6-12 IRS. In addition, we calculated the survival probability depending on the IRS (Figure 2). Patients with an IRS of 6 or higher have a similar bad prognosis compared to patients with an IRS below 5. A better survival correlates with lower MACC1 IRS. Patients with an IRS of 0 have the best prognosis. Based on these two analyses, the decision for cut-off value of 5 was made. Additionally, ROC curve analysis showed, that the IRS value of 5 has the highest Youden Index and is therefore the most suitable cut-off value. 0.408) (Figure 3 and Table 1). Looking at MACC1 expression in cell lines and tissue samples, the variability looks similar. Nevertheless, it must be considered that the investigation of the homogeneous cell lines is a purely quantitative analysis of the expression by Western plot and RT-PCR. The analysis of the tissue samples by immunoreactivity score analyses both the strength of expression and the inhomogeneity of expression. Figure 1: Distribution of the IRS across the cohort Figure 2: Overall Survival depending on the IRS Figure 3: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Table 1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	2	1	"""MACC1 is an independent negative prognostic marker in AGE/S patients."""			cancers14071773_perova	0
+We thank the reviewer for raising this point. However, to respond to this question appropriately, we currently can provide only the results from proliferation assays for one cell line model (FLO1-EV vs FLO1-MACC1) over the time period of 72 h. This assay was with no statistically significant difference in cell proliferation between FLO1-EV vs FLO1-MACC1 (see Figure 4). Figure 4: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay. Unfortunately, the short revision period of 10 days limits the generation of additional data sets for proliferation in further tumor cell lines.	2	1	[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249).			cancers14071773_perova	0
+We thank the reviewer for this question. The concept of MACC1 inhibition was originally developed by our group for the treatment of MACC1 positive colorectal cancers and was recently published (Kobelt et al. Oncogene 2021). In this study we transferred our experience with MACC1 inhibition by selumetinib on AGE/S. Therefore, we chose the same doses for in vitro and in vivo experiments, and which was also used by other authors (Huynh et al. Mol Cancer Therapy. 2007). Further, we knew that the selected selumetinib concentrations of 10 µM has no cytotoxic effect in vitro and showed no reduction of migration in the MACC1 negative clones (FLO1-EV and OEshMACC1). Regarding the different dosages in the context of xenografted mice treatment in vivo and the clinically used dosages, the FDA recommends multiplying the dosage by a factor of 0.08 when transferring from mouse to human (Niar et al. Journal of basic and clinical pharmacy 2016). According to this calculation, the dosage of 50 mg selumetinib /kg mouse used would equal a dosage of 4 mg/kg in humans. The clinical dose of 75mg twice daily corresponds to a dose of 2x1mg per kg assuming a patient weight of 75kg. Using the aforementioned calculation, this dose equals then 12.5 mg/kg for treatment of mice. In our study, we refer again to the work of Huynh et al. Here, in which a dosage of 12 mg/kg selumetinib showed only a low level of effectiveness. At 100 mg/kg, a very good effectiveness was seen, however combined with an increased toxicity. Thus, we used in our study the dosage of 50 mg/kg as an effective dosage with low toxicity for the mice. Overall, we are considering here a pre-clinical model. The possibility of MEK1 inhibition to inhibit MACC1-induced effects in AGE/S was examined. Further, extended in vivo studies might then reveal the ideal dosage in vivo, being thereafter translated to use in humans. The use of trametinib is an interesting suggestion and might well be adapted to its use in forthcoming studies References: Kobelt, D., Perez-Hernandez, D., Fleuter, C., Dahlmann, M., Zincke, F., Smith, J., ... & Stein, U. (2021). The newly identified MEK1 tyrosine phosphorylation target MACC1 is druggable by approved MEK1 inhibitors to restrict colorectal cancer metastasis. Oncogene, 40(34), 5286-5301. Huynh H, Soo KC, Chow PK, Tran E. Targeted inhibition of the extracellular signal-regulated kinase kinase pathway with AZD6244 (ARRY-142886) in the treatment of hepatocellular carcinoma. Mol Cancer Ther. 2007 Jan;6(1):138-46.. Nair, Anroop B.; Jacob, Shery. (2016). A simple practice guide for dose conversion between animals and human. Journal of basic and clinical pharmacy, 7. Jg., Nr. 2, S. 27.	2	1	Inhibition of MEK1 with selumetinib also reduced cell migration only in MACC1 expressing cells.			cancers14071773_perova	0
+We thank the reviewer for this comment. The IRS cut-off value 5 reaches a sensitivity of 76.6% and a specificity of 64.2% (Youden Index. We include the arguments into the methods section page 3: To define the cut-off value a ROC curve analysis was performed. 0.408) and was chose as cut-off value (see supplement Figure S1 and Table S1). Additionally, we added the ROC curve and the table with sensitivity, specificity and Youden Index into a supplement section as figure S1 and table S1 Figure S1: ROC Curve for IRS cut-off analysis (AUC 0.716 [0.656 – 0.776]) Positive if Greater than or Equal to Sensitivity 1- Specificity Specificity Youden Index -1 1 1 0 0 1 0,869 0,500 0,500 0,369 3 0,864 0,483 0,517 0,381 5 0,766 0,358 0,642 0,408 7 0,729 0,342 0,658 0,387 8,5 0,421 0,217 0,783 0,204 10,5 0,411 0,200 0,800 0,211 13 0 0 1 0 Table S1: Sensitivity, specificity and Youden Index for different IRS cut-off values.	4	1	I recommend the following edits before publication.			cancers14071773_perova	0
+thank you for this comment: we added the proliferation analyses by MTT into the method section page 5: Additionally, proliferation for FLO-1/EV and FLO-1/MACC1 was analyzed over 72 h by MTT. 4x103 cells were plated into 96-well-plates and were allowed to accommodate for 24 h. after 48 and 72 h formazan crystals were dissolved in 150 μl of DMSO and the absorption was measured at 560 nm in the absorbance reader (Tecan infinite 200 PRO). Each cell proliferation experiment was performed in triplicates. We added the results of the MTT analyses into the results section page 11: The additional analysis of the proliferation over 72 h showed no significant differences between FLO-1/EV (100% ± 4.886) and FLO-1/MACC1 (112.3% ± 12.24; p=0.113) (see supplement Figure S3). We added the MTT plot as figure S2 into the supplement section Figure S2: Proliferation of FLO1 EV and FLO1 MACC1 over 72 h. Analysis of in vitro proliferation assay.	4	1	[3] Throughout the paper, 10^x wrongly displays as 10x (e.g.			cancers14071773_perova	0
+We apologize for the inconvenience. Figure 4 can be found on page 14 in the submitted version. We inserted Figure 4 in the revised manuscript again to solve the technical issue.	2	1	Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 1:			cancers14071773_perova	0
+We address this question in the results part MACC1 expression: “ Due to the procedure of cutting and staining, some samples could not be used for the evaluation. Samples with insufficiently representative tumor tissue were also excluded from the evaluation. Cores with representative tumor material and evaluable staining were available in 266 of 360 samples (73.9%)…”. To avoid misunderstandings, we corrected the abstract to: 266 Samples of 360 AGE/S patients were analyzed for MACC1 expression.	2	1	It would be nice to know the selumetinib sensitivity of the two used cell lines.			cancers14071773_perova	0
+We thank the reviewer for this correction. This formatting issue was corrected throughout the entire manuscript.	2	1	The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.			cancers14071773_perova	0
+We thank the reviewer for this valuable hint. As suggested, we included p-values in Figure 3G and 3H. In Figure 3G there is a significant increase in migration of cells without MACC1 expression. This is based on a sufficiently large N, that renders even small differences statistically significant. Further, this increase is rather small and is outperformed by the MACC1-mediated increase in migration and the reduction in the MACC1-mediated migration by selumetinib. The data show, that selumetinib does not reduce migration of the MACC1-negative FLO1 and that MACC1 induces migration in this cell line that can be inhibited by selumetinib.	2	1	[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).			cancers14071773_perova	0
+We thank the reviewer for this suggestion to improve the visual quality of the manuscript. The legends of 3F and I have been enlarged and the resolution of the figure has been increased.	2	1	"""MACC1 is an independent negative prognostic marker in AGE/S patients."""			cancers14071773_perova	0
+We thank the reviewer for this discussion. The empty vector control clone was transduced and selected afterwards. During the selection procedure it is possible that a subclone was generated that expresses slightly more (or less) of any mRNA and protein. It differs from the wildtype by handling (transduction, selection etc) and somewhat by age (cell divisions). Therefore, for all comparisons the empty vector clone was used, as this clone was generated in parallel to the MACC1 clone.	2	1	"should be after ""Expression was correlated with survival and morphological characteristics."""			cancers14071773_perova	0
+We thank the reviewer for pointing this out: Survival reports were changed in median survival and the 95% confidence interval were included. Additionally, disease-specific survival results were reported.	2	1	"[8] In line 274, please change ""OR 1.51"" to ""OR 1.51 [... - ...]"", i.e."			cancers14071773_perova	0
+We thank the reviewer for pointing this out: OR with the confidence interval is now included.	2	1	[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values).			cancers14071773_perova	0
+We thank the reviewer for pointing this out: all survival reports were changed in median survival and the 95% confidence interval were included.	2	1	[6] In line 306/307, please explain/speculate briefly why the control clone had higher MACC1 expression than the wildtype.			cancers14071773_perova	0
+Thank you for this valuable comment. The journal requires an unstructured abstract. As suggested by the reviewer, we have therefore reorganized the abstract and improved the flow of reading: Abstract: Esophageal and Gastric Adenocarcinomas (AGE/S) are characterized by early metastasis and poor survival. MACC1 (Metastasis Associated in Colon Cancer 1) acts in colon cancer as a metastasis inducer, linked to reduced survival. This project illuminates the role and potential of inhibition of MACC1 in AGE/S. Using 266 of 360 TMAs and survival data of AGE/S patients we confirm the value of MACC1 as an independent negative prognostic marker in AGE/S patients. MACC1 gene expression correlated with survival and morphological characteristics. In vitro analysis of lentivirally MACC1 manipulated subclones of FLO-1 and OE33 showed enhanced migration induced by MACC1 in both cell line models, which could be inhibited by the MEK1 inhibitor selumetinib. In vivo, the efficacy of selumetinib on tumor growth and metastasis of MACC1-overexpressing FLO-1 cells xenografted intrasplenically in NOG mice was tested. Mice with high MACC1 expressing cells developed faster and larger distant metastases. Treatment with selumetinib led to a significant reduction of metastasis exclusively in the MACC1 positive xenografts.	2	1	Author Response Please see the attachment Author Response File: Author Response.pdf			cancers14071773_perova	0
+There were no more open questions in the second review.	4	1	[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249).			cancers14071773_perova	0
+We thank the reviewer for this important question and would be happy to clarify. TTFields are alternating electric fields in the range of 100 to 500 kHz, with maximal efficacy seen at a different frequency for different tumor types. Since TTFields cannot be applied at several different frequencies simultaneously, one specific frequency needs to be selected, and so the first step in examination of a new tumor type is a frequency scan. The effect of TTFields is not dichotomic, such that only one frequency is effective while the other are not at all. From the frequencies showing efficacy the purpose is to select the most effective one. It should be clarified that higher frequencies of the alternating electric fields do not mean higher energy, and hence there is no disadvantage in working with higher frequencies and no added toxicity or increased side effects for higher frequencies within the TTFields frequency range. Of note, TTFields at 200 kHz is already approved and has been applied to more than 18,000 patients with glioblastoma, with skin irritation being the main treatment related adverse event. TTFields at 150 kHz is approved for treatment of unresectable malignant pleural mesothelioma. To address these issues, we have rephrased the discussion: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” and “The lower TTFields intensity required for treatment of HepG2 relative to Huh-7D12 cells for achieving the same level of efficacy suggest higher sensitivity of the former to TTFields.” Point 2: What is the ‘n’ number of the samples and experiments in each group?	2	1	Majority of rodent study in cancer research lose the control group early but the treated groups are followed up for much longer to delineate indeed the efficacy vs. toxicity.			cancers14122959_makarova	0
+We thank the reviewer for pointing this was not clear. As mentioned in the statistical analysis sub-section within the methods part, all in vitro experiments were repeated at least 3 times, and so depicted values are mean (N ≥ 3) ± SEM. We have now added this also to the legends of figures 1, 2, and 3. Per the in vivo study, as was mentioned in the methods (sub-section 2.8), 52 animals were included in the study. The specific numbers in each treatment group are now mentioned in the results (sub-section 3.4): “During the treatment period, average tumor volumes of control animals (n = 11) increased 5.9-fold (Figure 4b and 4c; and Figure S3 for tumor images). For animals treated with TTFields (n = 15) or sorafenib (n = 10), tumor growth was significantly lower, 3.3- and 2.3-fold, respectively. In the TTFields-sorafenib combination group (n = 16), a 1.6-fold increase in tumor volume within the treatment period was observed, a growth significantly lower than that for control or for each treatment alone.” It is also mentioned in the legend of figure 4: “Rats (n = 52) were inoculated orthotopically with rat HCC N1S1 cells, and treated with sham-vehicle (n = 11), TTFields alone (n = 15), sorafenib alone (n = 10), or TTFields + sorafenib (n = 16), according to the depicted timeline (a).” Point 3: LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group.	2	1	Point 9: Was Mycoplasma testing done routinely?			cancers14122959_makarova	0
+First, we would like to emphasize that indications of autophagy were diverse and determined from increased cellular granularity, amplified lipidation of LC3 (from LC3-I to LC3-II) detected by Western blot, and elevated levels of LC3 foci observed by fluorescent staining. Nevertheless, to better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and additional time points. As per the reviewer’s request, these examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3.	2	1	As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib.			cancers14122959_makarova	0
+Per the reviewer’s request we show below images of the tumors within the liver, which we have also provided now in the supporting material of the paper. As may be seen, the liver itself is much larger than the tumor, and so we did not see value in measuring total liver weight, and hence cannot provide these values. While we tried to reduce variability between the animals by excluding those that at treatment initiation had tumor volumes not within the range 30–100 mm3, there was still divergence between the animals that resulted in the standard deviation bars depicted in the figure. It is however important to mention that there was a very good correlation between tumor volume (as measured by MRI) versus tumor weight at the end of the study, as may be seen in the graph below.	2	1	Point 10: Line 337 seems over statement since the data are not shown in the manuscript.			cancers14122959_makarova	0
+IHC tests the presence of a protein already expressed in the tissue, and 30 min are sufficient to allow recognition and binding of the antibody to the target protein. The secondary antibody is an HRP conjugated goat anti rabbit, that is a part of the Leica HRP-refine detection kit (Cat # DS9800). We have added the missing antibody information to section 2.10. We unfortunately do not have any tissue left to allow for performing WB analysis from the in vivo study, but we have conducted additional cell line studies and added WB analysis of cleaved PARP for the in vitro examinations and additional IHC examinations for autophagy and ER stress markers (as shown in response 3).	2	1	Response 3: We sincerely appreciate this well-taken comment.			cancers14122959_makarova	0
+We thank the reviewer for identifying this mistake, and have corrected this, changing the label of the LC3 data to e.	2	1	Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but only 2-fold in the combination group (Figure 4e).			cancers14122959_makarova	0
+Error bars are not missing for the control groups, rather they are null, as the values displayed in these figures are relative to control, and so per definition all control experiments have the exact same value of 100 or 1 (for percentage or fold change, respectively).	2	1	This study identified the efficacy of TTFields treatment for hepatocellular carcinoma and the combined effect of sorafenib in vitro and in vivo.			cancers14122959_makarova	0
+We apologize for this mistake, and have now corrected the figure legend. The quantification of immunofluorescence of LC3 foci formation is c, and immunoblotting showing LC3-II to LC3-I ratio is d. Point 9: There is no data in Fig 4A.	2	1	While I do not know if the authors have the technology to perform TTFields in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells.			cancers14122959_makarova	0
+We apologize for accidently omitting Fig 4A. This figure depicts the timeline of the in vivo experiment for easier understanding of the study designed. We have now corrected this.	2	1	Response 7: We thank the reviewer for this question.			cancers14122959_makarova	0
+We understand from the reviewer’s comments that the sentence “The higher effects seen in the presence of CQ reveal that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation” is not clear enough. We have hence rephrased the sentence to be more accurate and better deliver the message: “CQ is an inhibitor of lysosome degradation, commonly used to decipher whether the elevation of LC3 is due to upregulation of the autophagy process or reduced autophagosome turnover [30]. The higher TTFields-induced elevation of LC3 seen in the presence of CQ suggest that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation.” Point 11: Typos: Line 111, ‘invitro’ spelling is not correct.	2	1	What are the p53 status and the apoptosis signaling pathway function in N1S1 cells?			cancers14122959_makarova	0
+The “inovitro” in line 111 is not a typo, it is the name of the system used for applying TTFields in vitro. In line 342 we removed the duplicate “and”, and thank the reviewer for catching this typo.	2	1	*p < 0.05 relative to control; one-way ANOVA.			cancers14122959_makarova	0
+We thank the reviewer for the positive review.	2	1	I have raised the following concerns which is necessary to make this manuscript more scientifically interesting.			cancers14122959_makarova	0
+For the in vitro experiments with TTFields (sub-section 2.3) we tried not to elaborate too much and referenced previous work describing all details. We understand that we have cut off too many details, and are happy to add them “HepG2 and Huh-7D12 cell suspensions (500 µl, 25 x 103 cells/plate) were placed as a drop in the center of 35-mm inovitro™ dishes composed of high dielectric constant ceramic (lead magnesium niobate–lead titanate [PMN-PT]), with two perpendicularly pairs of transducer arrays printed on their outer walls. Cells were incubated overnight at 37 °C to allow attachment to the dish, and then 2 ml of fresh media were added.” Regarding the number of rats included in the final analysis, we apologize this information was not clear. The 52 rats mentioned in the text were in fact the actual final number of animals in the analysis, since “all rats reached the required usage limit of ≥18 h/day”, as is now explicitly stated in the results sub-section 3.4.” Point 2: What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells.	2	1	Author Response Please see attachment Author Response File: Author Response.docx			cancers14122959_makarova	0
+We thank the reviewer for raising this important issue. Using a xerograph model is indeed a good suggestion, however it would require working in immunosuppressed mice. It is currently not feasible for us to perform studies in which we apply TTFields to immunosuppressed animals, as these animals are of smaller weight thus the burden of wearing the electrodes may induce too much stress. Furthermore, these animals are more prone to contract infections during the process of electrodes placement and replacement on the animals. On the other hand, using the rat cells for in vitro experiments in the inovitro dishes turned out to be very technically challenging, as the cells were non-adherent, and so we could not pursue this important avenue. Nevertheless, we agree with the reviewer that there is a possibility that the optimal frequency may differ in the N1S1 relative to the human cell lines. Therefore, we have addressed this issue by including results of an additional frequency scan experiments done with N1S1 cells. We now mention this in the text, results sub-section 3.4: “The efficacy of combining TTFields with sorafenib relative to each modality alone was examined in the N1S1 HCC rat orthotopic model (timeline in Figure 4a). In vitro experiments confirmed that the 150 kHz TTFields frequency found optimal for treatment of the human cell lines was also optimal for treatment of the murine N1S1 cells used for the in vivo study (Figure S2).” We provide the relevant frequency scan figure in the supplementary material. Per the good question by the reviewer regarding the p53 status of the N1S1 cells we added: “It is also worth mentioning that the N1S1 murine cells used for this study, like the HepG2 cells, are p53 wild type.” Figure S2.	2	1	HepG2 cells were treated for 6, 24, or 48 hours with 150 kHz TTFields, 3 µM sorafenib, or the two treatments combined, followed by Western blot examination of the autophagy markers beclin-1 and LC3 (d), the ER stress marker GRP78 (e), and the apoptosis marker cleaved PARP (f).			cancers14122959_makarova	0
+We thank the reviewer for this comment, and apologize that the illustration of the timeline was accidentally missing from the submission (we have now added it). The limiting factor for study duration was the well being of the animals. The tumors in the control group were very large, causing stress and weight loss of the animals. The arrays placed on the animals, together with the individual housing needed to prevent wire entanglement, adds even more stress and increases animal weight loss. Overall, it was non-ethical to continue the study further. We have now added this explanation to the discussion section to clarify this limitation of the study: “In the HCC animal model, the acute effects of TTFields and sorafenib were examined. Due to the large tumors developed in the control group and the stress experienced by the animals as a result of the individual housing and motility limitations imposed by the sham and TTFields arrays, longer treatment durations were not feasible.” Point 4: There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing.	2	1	It is practically impossible to appreciate a therapeutic effect in such a short time and upon monitoring effect for 1 day.			cancers14122959_makarova	0
+It is important to understand that the TTFields frequency is not the treatment dose, and hence increased frequency does not mean increased efficacy. In the case of TTFields, intensity, duration, and usage are the factors that contribute to the “treatment dose”. Nevertheless, we have added to the discussion an explanation on this issue of optimal frequency: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” Regarding the option that the optimal frequency will differ between in vitro and in vivo setting, this is indeed a concern. However, as shown by the in vitro frequency scans, the effect of TTFields is not dichotomic, and effectiveness can be seen at more than one frequency. Therefore, the relatively low expected differences between animals treated with different TTFields frequencies together with the variability inherent to animal studies, makes it technically problematic to perform frequency scans in vivo. As of today, in vitro frequency scans are the common practice for determining the optimal TTFields frequency for delivery to animals and to humans. In fact, in vitro frequency scans were the basis for the clinical studies leading to FDA approval of TTFields at 200 kHz for the treatment of GBM and at 150 kHz for the treatment of MPM. We have added this issue to the discussion as a limitation of the study: “Since the optimal TTFields frequency has been shown to be dependent on the electrical properties of the cells and it is not clear how much effect the tumor microenvironment has on these properties, and because it is technically problematic to perform TTFields frequency scans in vivo, the frequency detected in the cell cultures was also employed for the animal studies.” Point 5: Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	2	1	When compared with sorafenib alone there is practically no difference.			cancers14122959_makarova	0
+When it comes to testing efficacy, we would like to use a treatment duration as long as possible to maximize the effect. However, this duration varies between in vitro and in vivo work. For cell cultures, the control samples continue to grow throughout the treatment duration, and so after 72 h the plates are very much confluent, and may not be left to continue and grow any further (without drastic changes in the environmental conditions, making them inadequate control cells). It is also noteworthy that such treatment periods for in vitro work are well accepted in the field of TTFields research. In vivo, prior studies also used similar time frames of 2 to 3 weeks from inoculation when working with the N1S1 model (Buijs et al., 2012 -; Ju et al., 2009; Thompson et al., 2012). Extending treatment duration was also limited by the physical status of the animals, as was explained in response 3.	2	1	N1S1 cells were treated with TTFields (1.7 V/cm RMS) across a frequency range of 100–400 kHz, and cell count were determined following 24 hours of treatment.			cancers14122959_makarova	0
+Chloroquine was added to the cell cultures only at the final hours of the treatment for answering questions related to the mechanism of action, and not for boosting the efficacy of the other treatments. When using chloroquine in animal studies, it is for efficacy purposes, and so it is used throughout the treatment period. Since our animal experiment aimed to examine the efficacy of concomitant TTFields and sorafenib, with no additional agents, chloroquine was not employed.	2	1	Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic?			cancers14122959_makarova	0
+For the cytotoxicity assay we remove the supernatant, washed the cells, and then collect the adherent cells following trypsinization and visual inspection to verify all cells were removed. Indeed, as the reviewer mentioned correctly, we “counted the live cells and plotted the final data as percentage of untreated controls”. We agree that this is not cytotoxicity per se, and that is why we also perform 7-AAD/annexin-V staining of the cells. For this apoptosis assay we do collect the supernatant together with the adherent cells. Increased apoptosis and/or necrosis indicates that reduction in the cell number observed in the cell count emanates at least in part from cytotoxicity. We agree with the reviewer that we should not confuse the cell count measurements with the term “cytotoxicity” prior to showing the effect on apoptosis, and therefore we have changed this terminology throughout the paper.	2	1	Maybe the cytotoxicity data will reveal a different better frequency.			cancers14122959_makarova	0
+We thank the reviewer for these questions. Preliminary tests using CD31 staining revealed no differences between the groups regarding blood vessel density and therefore we did not pursue the research in that direction. We agree that measuring the effect of TTFields on the anti-angiogenic effect of sorafenib and on resistance to sorafenib are interesting, these topics were however not within the scope of the current study, and remain for future investigations. Regarding possible discrepancy between the volume fold change and tumor weight, please see below a graph showing tumor volume (as measured by MRI) versus tumor weight, both measured at the end of the study. The graph shows very good correlation between the two parameters, indicating the reliability of the measurements. If the reviewer feels there is discrepancy, it may be due to the volume shown as fold change relative to the initial tumor volume and not as the end value.	2	1	Response 3: We sincerely appreciate this well-taken comment.			cancers14122959_makarova	0
+We routinely examine our cells for Mycoplasma. Regarding authenticity, the cells were used shortly after purchase, and so there was no need to examine this.	2	1	Point 10: Line 337 seems over statement since the data are not shown in the manuscript.			cancers14122959_makarova	0
+We appreciate this question. Autophagy is a process that is elevated in order to cope with cellular stress, but as stress level elevate, autophagy can no longer provide the protection the cell needs, and the cell will undergo apoptosis. This kind of kinetics means that the levels of autophagy markers will depend on the time point the cells are examined. This may be appreciated from figure 2 panel c and d, with the different kinetics displayed by the two cell lines. While for Huh-7D12 autophagy levels increase from 24 to 48 hours of treatment, in the HepG2 cells autophagy levels at 48 hours are lower than at 24 hours, indicating these cells are already after the autophagy peak and on the way to apoptosis. Indeed, figure 1d shows higher levels of apoptosis for HepG2 cells. To clarify we have added a few sentences to the text. In results sub-section 3.2: “However, autophagy kinetics seems to be faster in the HepG2 cells, in which LC3 markers are lower at 48 versus 24 hours, whereas elevation is seen from 24 to 48 hours for the Huh-7D12 cells.” In the discussion “While autophagy serves as a survival strategy of cells, when stress levels continue raising it may be over-activated and mediate cell death [9]. The faster autophagy kinetics seen for the HepG2 relative to Huh-7D12 cells following application of TTFields is in agreement with the higher apoptosis levels displayed by this cell line, and may serve as an additional rational for the higher efficacy of TTFields against it. Examination of the reasons for faster autophagy in HepG2 relative to Huh-7D12 cells is out of the scope of this work.” For more clarity we have also added a more in-depth kinetic study, including additional relevant markers and additional time points, examinations that were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case” Figure 3.	2	1	*p < 0.05, **p < 0.01, and ****p < 0.0001 relative to control for labels above bars, or between indicated groups; Student’s T-test.			cancers14122959_makarova	0
+The sentence referred to was meant to describe only the in vivo outcomes. We have rephrased it to be more accurate and clear: “While each treatment alone elevated levels of autophagy relative to control, TTFields concomitant with sorafenib induced a significant increase versus control in tumor ER stress and apoptosis levels, demonstrating increased stress under the multimodal treatment.” Point 12: Finally, adding to all the above questions, I found a very weak Discussion section which must be extended.	2	1	GRP78 levels in the groups treated with TTFields or sorafenib alone remained unchanged from the control, but were elevated 2-fold in the TTFields plus sorafenib group (Figure 4f).			cancers14122959_makarova	0
+We thanks the reviewer for this comment. We have now elaborated on many issues throughout the discussion, as was described thorough this response letter. Regarding the conclusion, we have rephrased it for better accuracy: “TTFields were identified to be most efficient for treatment of HCC cells at 150 kHz, and this frequency further demonstrated in vivo efficacy.” Why only one frequency was used in vivo, and the difference between frequency and dose, were explained in response 4.	2	1	There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.			cancers14122959_makarova	0
+We thank the reviewer for this question. Indeed, the sorafenib dose used in this study proved to be slightly more efficacious than TTFields in controlling tumor fold increase. Nevertheless, these differences between the monotherapies, did not reach statistical significance. In accordance, there was no statistical difference between the monotherapies in the expression levels of the LC3 marker and the levels of cleaved PARP. In order to better understand the mechanism of action, we have added experiments to better characterize the autophagy-apoptosis interplay for treatment with concomitant TTFields and sorafenib, which are described in response 3. Of note, the clinical development of TTFields does not aim to replace sorafenib with TTFields, but rather to add TTFields on top of sorafenib and therefore this work focused mainly on the potential added value in combining these 2 modalities.	2	1	*p < 0.05, **p < 0.01, and ****p < 0.0001 relative to control for labels above bars, or between indicated groups; Student’s T-test.			cancers14122959_makarova	0
+We thank the reviewer for pointing out this issue. Quantification of the IHC images was done automatically. The whole slide was scanned, and the CaseViewer software was used to exclude non-tumor areas. The signals of the stained protein and the nuclei were resolved by color deconvolution and quantified separately using the FIJI software (ImageJ) software. Average signal per cell or percent of positive cells was calculated. As the reviewer pointed out, the high magnification images we chose to show do not correctly reflect the quantification performed by the software, and we have now replaced them with better representative fields of the slides.	2	1	Images of tumors within the livers of the rats treated with sham heat (control rats), TTFields, sorafenib, or TTFields plus sorafenib.			cancers14122959_makarova	0
+We sincerely appreciate this well-taken comment. To better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and at additional time points. These examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3.	2	1	 Point 7: Why there is no error bar in the control group of all bar graphs?			cancers14122959_makarova	0
+Both cell lines experience elevation of apoptosis following application of sorafenib in a dose dependent manner, as evident from the AnnV/7AAD results. However, while TTFields greatly elevate apoptosis in HepG2 cells, they have a low effect on apoptosis levels in the Huh-7D12 cells, seen both in Figure 1d and in Figure 3c. As was explained in the discussion, this difference between the cell lines may be attributed to the different p53 status, wild type in HepG2 and mutated in Huh-7D12, as there are previous indications of lower TTFields-induced apoptosis in cell lines with mutated p53. As suggested by the reviewer, in order to back up the AnnV/7AAD results we added WB for cleaved PARP, as described in response 3.	2	1	Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			cancers14122959_makarova	0
+We apologize for accidentally leaving out this figure, and have now added it.	2	1	Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).			cancers14122959_makarova	0
+We thank the reviewer for this comment. In the in vitro experiments we used cell lines derived from humans. However, these cell lines cannot be implanted to rats, and so for the in vivo experiments we had to use a cell line from rats.	2	1	While for in vitro data, the authors have data, for in vivo they used only one frequency of 150 Hz.			cancers14122959_makarova	0
+We thank the reviewer for this important question and would be happy to clarify. TTFields are alternating electric fields in the range of 100 to 500 kHz, with maximal efficacy seen at a different frequency for different tumor types. Since TTFields cannot be applied at several different frequencies simultaneously, one specific frequency needs to be selected, and so the first step in examination of a new tumor type is a frequency scan. The effect of TTFields is not dichotomic, such that only one frequency is effective while the other are not at all. From the frequencies showing efficacy the purpose is to select the most effective one. It should be clarified that higher frequencies of the alternating electric fields do not mean higher energy, and hence there is no disadvantage in working with higher frequencies and no added toxicity or increased side effects for higher frequencies within the TTFields frequency range. Of note, TTFields at 200 kHz is already approved and has been applied to more than 18,000 patients with glioblastoma, with skin irritation being the main treatment related adverse event. TTFields at 150 kHz is approved for treatment of unresectable malignant pleural mesothelioma. To address these issues, we have rephrased the discussion: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” and “The lower TTFields intensity required for treatment of HepG2 relative to Huh-7D12 cells for achieving the same level of efficacy suggest higher sensitivity of the former to TTFields.” Point 2: What is the ‘n’ number of the samples and experiments in each group?	2	1	The manuscript is an interesting work related to a potential new therapy of hepatocellular carcinoma (HCC) by using TTFields in combination with a TKI (Sorafenib).			cancers14122959_perova	0
+We thank the reviewer for pointing this was not clear. As mentioned in the statistical analysis sub-section within the methods part, all in vitro experiments were repeated at least 3 times, and so depicted We have now added this also to the legends of figures 1, 2, and 3. Per the in vivo study, as was mentioned in the methods (sub-section 2.8), 52 animals were included in the study. The specific numbers in each treatment group are now mentioned in the results (sub-section 3.4): “During the treatment period, average tumor volumes of control animals (n = 11) increased 5.9-fold (Figure 4b and 4c; and Figure S3 for tumor images). For animals treated with TTFields (n = 15) or sorafenib (n = 10), tumor growth was significantly lower, 3.3- and 2.3-fold, respectively. In the TTFields-sorafenib combination group (n = 16), a 1.6-fold increase in tumor volume within the treatment period was observed, a growth significantly lower than that for control or for each treatment alone.” It is also mentioned in the legend of figure 4: “Rats (n = 52) were inoculated orthotopically with rat HCC N1S1 cells, and treated with sham-vehicle (n = 11), TTFields alone (n = 15), sorafenib alone (n = 10), or TTFields + sorafenib (n = 16), according to the depicted timeline (a).” Point 3: LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group.	2	1	The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment.			cancers14122959_perova	0
+First, we would like to emphasize that indications of autophagy were diverse and determined from increased cellular granularity, amplified lipidation of LC3 (from LC3-I to LC3-II) detected by Western blot, and elevated levels of LC3 foci observed by fluorescent staining. Nevertheless, to better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and additional time points. As per the reviewer’s request, Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but elevated only 2-fold in the combination group (Figure 4e).	2	1	The in vivo model appears to be performed only time which question the validity of data.			cancers14122959_perova	0
+Per the reviewer’s request we show below images of the tumors within the liver, which we have also provided now in the supporting material of the paper. As may be seen, the liver itself is much larger than the tumor, and so we did not see value in measuring total liver weight, and hence cannot provide these values. While we tried to reduce variability between the animals by excluding those that at treatment initiation had tumor volumes not within the range 30–100 mm3, there was still divergence between the animals that resulted in the standard deviation bars depicted in the figure. It is however important to mention that there was a very good correlation between tumor volume (as measured by MRI) versus tumor weight at the end of the study, as may be seen in the graph below. Figure S3. Images of tumors within the livers of the rats treated with sham heat (control rats), TTFields, sorafenib, or TTFields plus sorafenib.	2	1	There is no way to analyze bands for a protein using the GAPDH data from another experiment.			cancers14122959_perova	0
+IHC tests the presence of a protein already expressed in the tissue, and 30 min are sufficient to allow recognition and binding of the antibody to the target protein. The secondary antibody is an HRP conjugated goat anti rabbit, that is a part of the Leica HRP-refine detection kit (Cat # DS9800). We have added the missing antibody information to section 2.10. We unfortunately do not have any tissue left to allow for performing WB analysis from the in vivo study, but we have conducted additional cell line studies and added WB analysis of cleaved PARP for the in vitro examinations and additional IHC examinations for autophagy and ER stress markers (as shown in response 3).	2	1	Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but only 2-fold in the combination group (Figure 4e).			cancers14122959_perova	0
+We thank the reviewer for identifying this mistake, and have corrected this, changing the label of the LC3 data to e.	2	1	Kaplan-Meyer survival curves are mandatory when toxic treatments are combined and compared.			cancers14122959_perova	0
+Error bars are not missing for the control groups, rather they are null, as the values displayed in these figures are relative to control, and so per definition all control experiments have the exact same value of 100 or 1 (for percentage or fold change, respectively).	2	1	This way to present the data questions a lot the quality of experimental work and consequently and importantly the final analysis of data and the concluion claimed by the authors.			cancers14122959_perova	0
+We apologize for this mistake, and have now corrected the figure legend. The quantification of immunofluorescence of LC3 foci formation is c, and immunoblotting showing LC3-II to LC3-I ratio is d. Point 9: There is no data in Fig 4A.	2	1	Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group.			cancers14122959_perova	0
+We apologize for accidently omitting Fig 4A. This figure depicts the timeline of the in vivo experiment for easier understanding of the study designed. We have now corrected this.	2	1	Point 3: It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenib induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.			cancers14122959_perova	0
+We understand from the reviewer’s comments that the sentence “The higher effects seen in the presence of CQ reveal that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation” is not clear enough. We have hence rephrased the sentence to be more accurate and better deliver the message: “CQ is an inhibitor of lysosome degradation, commonly used to decipher whether the elevation of LC3 is due to upregulation of the autophagy process or reduced autophagosome turnover [30]. The higher TTFields-induced elevation of LC3 seen in the presence of CQ suggest that the observed phenomenon is due to increased autophagic flux, rather than decreased autolysosome degradation.” Point 11: Typos: Line 111, ‘invitro’ spelling is not correct.	2	1	This way to present the data questions a lot the quality of experimental work and consequently and importantly the final analysis of data and the concluion claimed by the authors.			cancers14122959_perova	0
+The “inovitro” in line 111 is not a typo, it is the name of the system used for applying TTFields in vitro. In line 342 we removed the duplicate “and”, and thank the reviewer for catching this typo.	2	1	The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d).			cancers14122959_perova	0
+For the in vitro experiments with TTFields (sub-section 2.3) we tried not to elaborate too much and referenced previous work describing all details. We understand that we have cut off too many details, and are happy to add them “HepG2 and Huh-7D12 cell suspensions (500 µl, 25 x 103 cells/plate) were placed as a drop in the center of 35-mm inovitro™ dishes composed of high dielectric constant ceramic (lead magnesium niobate–lead titanate [PMN-PT]), with two perpendicularly pairs of transducer arrays printed on their outer walls. Cells were incubated overnight at 37 °C to allow attachment to the dish, and then 2 ml of fresh media were added.” Regarding the number of rats included in the final analysis, we apologize this information was not clear. The 52 rats mentioned in the text were in fact the actual final number of animals in the analysis, since “all rats reached the required usage limit of ≥18 h/day”, as is now explicitly stated in the results sub-section 3.4.” Point 2: What is puzzling in this investigation is showing functional data using two human cell lines and in vivo data using rat cells.	2	1	Response: We thank the reviewer for the positive review.			cancers14122959_perova	0
+We thank the reviewer for raising this important issue. Using a xerograph model is indeed a good suggestion, however it would require working in immunosuppressed mice. It is currently not feasible for us to perform studies in which we apply TTFields to immunosuppressed animals, as these animals are of smaller weight thus the burden of wearing the electrodes may induce too much stress. Furthermore, these animals are more prone to contract infections during the process of electrodes placement and replacement on the animals. On the other hand, using the rat cells for in vitro experiments in the inovitro dishes turned out to be very technically challenging, as the cells were non-adherent, and so we could not pursue this important avenue. Nevertheless, we agree with the reviewer that there is a possibility that the optimal frequency may differ in the N1S1 relative to the human cell lines. Therefore, we have addressed this issue by including results of an additional frequency scan experiments done with N1S1 cells. We now mention this in the text, results sub-section 3.4: “The efficacy of combining TTFields with sorafenib relative to each modality alone was examined in the N1S1 HCC rat orthotopic model (timeline in Figure 4a). In vitro experiments confirmed that the 150 kHz TTFields frequency found optimal for treatment of the human cell lines was also optimal for treatment of the murine N1S1 cells used for the in vivo study (Figure S2).” We provide the relevant frequency scan figure in the supplementary material. Per the good question by the reviewer regarding the p53 status of the N1S1 cells we added: “It is also worth mentioning that the N1S1 murine cells used for this study, like the HepG2 cells, are p53 wild type.” Figure S2. TTFields frequency scan in rat N1S1 HCC cells. N1S1 cells were treated with TTFields (1.7 V/cm RMS) across a frequency range of 100–400 kHz, and cell count were determined following 24 hours of treatment. *p < 0.05 relative to control; one-way ANOVA. ANOVA = analysis of variance; HCC = hepatocellular carcinoma; RMS = root mean square; SEM = standard error of the mean; TTFields = Tumor Treating Fields.	2	1	Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.			cancers14122959_perova	0
+We thank the reviewer for this comment, and apologize that the illustration of the timeline was accidentally missing from the submission (we have now added it). The limiting factor for study duration was the well being of the animals. The tumors in the control group were very large, causing stress and weight loss of the animals. The arrays placed on the animals, together with the individual housing needed to prevent wire entanglement, adds even more stress and increases animal weight loss. Overall, it was non-ethical to continue the study further. We have now added this explanation to the discussion section to clarify this limitation of the study: “In the HCC animal model, the acute effects of TTFields and sorafenib were examined. Due to the large tumors developed in the control group and the stress experienced by the animals as a result of the individual housing and motility limitations imposed by the sham and TTFields arrays, longer treatment durations were not feasible.” Point 4: There is no explanation why the most effective dose was 150 kHz and higher does actually decreases the killing.	2	1	Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic?			cancers14122959_perova	0
+It is important to understand that the TTFields frequency is not the treatment dose, and hence increased frequency does not mean increased efficacy. In the case of TTFields, intensity, duration, and usage are the factors that contribute to the “treatment dose”. Nevertheless, we have added to the discussion an explanation on this issue of optimal frequency: “It has been previously shown that maximal effectivity of TTFields occurs at a different frequency for different cancer types, owing to the specific electrical properties of the cells [15, 28]. Hence, the first step in applying TTFields to a new tumor type includes frequency scans.” Regarding the option that the optimal frequency will differ between in vitro and in vivo setting, this is indeed a concern. However, as shown by the in vitro frequency scans, the effect of TTFields is not dichotomic, and effectiveness can be seen at more than one frequency. Therefore, the relatively low expected differences between animals treated with different TTFields frequencies together with the variability inherent to animal studies, makes it technically problematic to perform frequency scans in vivo. As of today, in vitro frequency scans are the common practice for determining the optimal TTFields frequency for delivery to animals and to humans. In fact, in vitro frequency scans were the basis for the clinical studies leading to FDA approval of TTFields at 200 kHz for the treatment of GBM and at 150 kHz for the treatment of MPM. We have added this issue to the discussion as a limitation of the study: “Since the optimal TTFields frequency has been shown to be dependent on the electrical properties of the cells and it is not clear how much effect the tumor microenvironment has on these properties, and because it is technically problematic to perform TTFields frequency scans in vivo, the frequency detected in the cell cultures was also employed for the animal studies.” Point 5: Why the in vitro experiment was performed for 72 hours and in in vivo for 120 hours?	2	1	In general, the manuscript is well written and provides novel and interesting data illustrating the possibility to enhance the efficacy of sorafenib in the therapy of hepatocellular carcinoma (HCC).			cancers14122959_perova	0
+When it comes to testing efficacy, we would like to use a treatment duration as long as possible to maximize the effect. However, this duration varies between in vitro and in vivo work. For cell cultures, the control samples continue to grow throughout the treatment duration, and so after 72 h the plates are very much confluent, and may not be left to continue and grow any further (without drastic changes in the environmental conditions, making them inadequate control cells). It is also noteworthy that such treatment periods for in vitro work are well accepted in the field of TTFields research. In vivo, prior studies also used similar time frames of 2 to 3 weeks from inoculation when working with the N1S1 model (Buijs et al., 2012 -; Ju et al., 2009; Thompson et al., 2012). Extending treatment duration was also limited by the physical status of the animals, as was explained in response 3.	2	1	Response 4: We thank the reviewer for this question.			cancers14122959_perova	0
+Chloroquine was added to the cell cultures only at the final hours of the treatment for answering questions related to the mechanism of action, and not for boosting the efficacy of the other treatments. When using chloroquine in animal studies, it is for efficacy purposes, and so it is used throughout the treatment period. Since our animal experiment aimed to examine the efficacy of concomitant TTFields and sorafenib, with no additional agents, chloroquine was not employed.	2	1	However, the limited experimental design and paucity of strong data ask for more experiments to proof the feasibility of this combination for treating HCC.			cancers14122959_perova	0
+For the cytotoxicity assay we remove the supernatant, washed the cells, and then collect the adherent cells following trypsinization and visual inspection to verify all cells were removed. Indeed, as the reviewer mentioned correctly, we “counted the live cells and plotted the final data as percentage of untreated controls”. We agree that this is not cytotoxicity per se, and that is why we also perform 7-AAD/annexin-V staining of the cells. For this apoptosis assay we do collect the supernatant together with the adherent cells. Increased apoptosis and/or necrosis indicates that reduction in the cell number observed in the cell count emanates at least in part from cytotoxicity. We agree with the reviewer that we should not confuse the cell count measurements with the term “cytotoxicity” prior to showing the effect on apoptosis, and therefore we have changed this terminology throughout the paper.	2	1	It looks like a speculative discussion section with no clear point of view of the authors and general conclusions of their presented work.			cancers14122959_perova	0
+We thank the reviewer for these questions. Preliminary tests using CD31 staining revealed no differences between the groups regarding blood vessel density and therefore we did not pursue the research in that direction. We agree that measuring the effect of TTFields on the anti-angiogenic effect of sorafenib and on resistance to sorafenib are interesting, these topics were however not within the scope of the current study, and remain for future investigations. Regarding possible discrepancy between the volume fold change and tumor weight, please see below a graph showing tumor volume (as measured by MRI) versus tumor weight, both measured at the end of the study. The graph shows very good correlation between the two parameters, indicating the reliability of the measurements. If the reviewer feels there is discrepancy, it may be due to the volume shown as fold change relative to the initial tumor volume and not as the end value.	2	1	Kaplan-Meyer survival curves are mandatory when toxic treatments are combined and compared.			cancers14122959_perova	0
+We routinely examine our cells for Mycoplasma. Regarding authenticity, the cells were used shortly after purchase, and so there was no need to examine this.	2	1	This is an interesting scientific study, as it concerns the issue of hepatocellular carcinoma, it also has a clinical aspect.			cancers14122959_perova	0
+We appreciate this question. Autophagy is a process that is elevated in order to cope with cellular stress, but as stress level elevate, autophagy can no longer provide the protection the cell needs, and the cell will undergo apoptosis. This kind of kinetics means that the levels of autophagy markers will depend on the time point the cells are examined. This may be appreciated from figure 2 panel c and d, with the different kinetics displayed by the two cell lines. While for Huh-7D12 autophagy levels increase from 24 to 48 hours of treatment, in the HepG2 cells autophagy levels at 48 hours are lower than at 24 hours, indicating these cells are already after the autophagy peak and on the way to apoptosis. Indeed, figure 1d shows higher levels of apoptosis for HepG2 cells. To clarify we have added a few sentences to the text. In results sub-section 3.2: “However, autophagy kinetics seems to be faster in the HepG2 cells, in which LC3 markers are lower at 48 versus 24 hours, whereas elevation is seen from 24 to 48 hours for the Huh-7D12 cells.” In the discussion “While autophagy serves as a survival strategy of cells, when stress levels continue raising it may be over-activated and mediate cell death [9]. The faster autophagy kinetics seen for the HepG2 relative to Huh-7D12 cells following application of TTFields is in agreement with the higher apoptosis levels displayed by this cell line, and may serve as an additional rational for the higher efficacy of TTFields against it. Examination of the reasons for faster autophagy in HepG2 relative to Huh-7D12 cells is out of the scope of this work.” For more clarity we have also added a more in-depth kinetic study, including additional relevant markers and additional time points, examinations that were performed for the combined treatment as compared to TTFields and sorafenib alone. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case” Figure 3. In the animal study we may only measure one time point. The lower autophagy seen at this time for the combined treatment together with the higher apoptosis indicate that we are further along the kinetic timeline of autophagy relative to the monotherapies, suggesting higher stress in animals receiving the combined treatment. We have now added IHC examination of beclin-1, an additional autophagy marker, and of GRP78, a marker for ER stress, as described in results sub-section 3.4.	2	1	Point 9: Was Mycoplasma testing done routinely?			cancers14122959_perova	0
+The sentence referred to was meant to describe only the in vivo outcomes. We have rephrased it to be more accurate and clear: “While each treatment alone elevated levels of autophagy relative to control, TTFields concomitant with sorafenib induced a significant increase versus control in tumor ER stress and apoptosis levels, demonstrating increased stress under the multimodal treatment.” Point 12: Finally, adding to all the above questions, I found a very weak Discussion section which must be extended.	2	1	Point 4: Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib (when compared to the cells treated with TTFields and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 (for both HCC cell lines).			cancers14122959_perova	0
+We thanks the reviewer for this comment. We have now elaborated on many issues throughout the discussion, as was described thorough this response letter. Regarding the conclusion, we have rephrased it for better accuracy: “TTFields were identified to be most efficient for treatment of HCC cells at 150 kHz, and this frequency further demonstrated in vivo efficacy.” Why only one frequency was used in vivo, and the difference between frequency and dose, were explained in response 4.	2	1	Response 6: We thank the reviewer for this question.			cancers14122959_perova	0
+We thank the reviewer for this question. Indeed, the sorafenib dose used in this study proved to be slightly more efficacious than TTFields in controlling tumor fold increase. Nevertheless, these differences between the monotherapies, did not reach statistical significance. In accordance, there was no statistical difference between the monotherapies in the expression levels of the LC3 marker and the levels of cleaved PARP. In order to better understand the mechanism of action, we have added experiments to better characterize the autophagy-apoptosis interplay for treatment with concomitant TTFields and sorafenib, which are described in response 3. Of note, the clinical development of TTFields does not aim to replace sorafenib with TTFields, but rather to add TTFields on top of sorafenib and therefore this work focused mainly on the potential added value in combining these 2 modalities.	2	1	These examinations were performed for the combined treatment as compared to TTFields and sorafenib alone.			cancers14122959_perova	0
+We thank the reviewer for pointing out this issue. Quantification of the IHC images was done automatically. The whole slide was scanned, and the CaseViewer software was used to exclude non-tumor areas. The signals of the stained protein and the nuclei were resolved by color deconvolution and quantified separately using the FIJI software (ImageJ) software. Average signal per cell or percent of positive cells was calculated. As the reviewer pointed out, the high magnification images we chose to show do not correctly reflect the quantification performed by the software, and we have now replaced them with better representative fields of the slides.	2	1	Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			cancers14122959_perova	0
+values are mean (N ≥ 3) ± SEM. We sincerely appreciate this well-taken comment. these examinations were performed for the combined treatment as compared to TTFields and sorafenib alone. We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results. These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins. For HepG2 cells, the autophagy marker beclin-1 demonstrated elevation after 6 hours of treatment, which was later replaced with diminished expression levels (Figure 3d). This type of behavior was seen in all treatment groups, but was most pronounced for TTFields-sorafenib co-application. The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d). As in the case of beclin-1, the magnitude of the effect was higher for co-treatment of TTFields and sorafenib relative to the monotherapies. GRP78, a marker of ER stress, remained low in all treatment groups for 6 and 24 hours of treatment, but demonstrated elevated levels at the later, 48-hours time point (Figure 3e). The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment. For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f). The slower kinetics of the autophagy-apoptosis path in the Huh-7D12 cells, as seen from the elevation of LC3 after as much as 48 hours (Figure 2c and d), prevented from detecting such changes in the levels of these markers in this cell line (Figure S1).” And in the discussion part “Kinetic examination in the HepG2 cells revealed elevation in autophagy levels as early as 6 hours of TTFields or sorafenib treatment, which diminished and were replaced with ER stress and apoptosis for 48 hours of treatment. These results are in line with a previous study that focused on the effects of sorafenib on such markers in HepG2 cells [32]. The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3. HepG2 cells were treated for 6, 24, or 48 hours with 150 kHz TTFields, 3 µM sorafenib, or the two treatments combined, followed by Western blot examination of the autophagy markers beclin-1 and LC3 (d), the ER stress marker GRP78 (e), and the apoptosis marker cleaved PARP (f). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA. In the animal study we have now added IHC examination of beclin-1 and of GRP78, a marker for ER stress, as described in results sub-section 3.4. Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively. GRP78 levels in the groups treated with TTFields or sorafenib alone remained unchanged from the control, but were elevated 2-fold in the TTFields plus sorafenib group (Figure 4f). Additionally, the percentage of cells positive for cleaved PARP was significantly higher relative to control only in the combination group (Figure 4g).” and also in the discussion: “The lower autophagy accompanied by the higher ER stress and apoptosis displayed in the conjunction group relative to the monotherapies groups following 6 days of treatment suggest that these animals were pushed further along the autophagy-apoptosis kinetic timeline due to the higher levels of stress experienced by these animals, in accordance with the results described for the cell cultures.” Figure 4. tumor slices were subjected to immunohistochemical analysis for beclin-1 and LC3 (e), GRP78 (f), and cleaved PARP (g). Values are mean ± SD. *p < 0.05, **p < 0.01, and ****p < 0.0001 relative to control for labels above bars, or between indicated groups; Student’s T-test. Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but only 2-fold in the combination group (Figure 4e). To better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and at additional time points.	2	1	I am not sure why the randomization of rats was unequally, practically 50% in TTF groups vs. control and sorafenib alone.			cancers14122959_perova	0
+Both cell lines experience elevation of apoptosis following application of sorafenib in a dose dependent manner, as evident from the AnnV/7AAD results. However, while TTFields greatly elevate apoptosis in HepG2 cells, they have a low effect on apoptosis levels in the Huh-7D12 cells, seen both in Figure 1d and in Figure 3c. As was explained in the discussion, this difference between the cell lines may be attributed to the different p53 status, wild type in HepG2 and mutated in Huh-7D12, as there are previous indications of lower TTFields-induced apoptosis in cell lines with mutated p53. As suggested by the reviewer, in order to back up the AnnV/7AAD results we added WB for cleaved PARP, as described in response 3.	2	1	Point 7: The authors claim that cytotoxicity was measured “by cell counting using iCyt EC800 (Sony Biotechnology) 123 flow cytometer, and expressed as a percentage relative to the control.” Does this imply that they counted the live cells and plotted the final data as percentage of untreated controls (as figure 1 suggests).			cancers14122959_perova	0
+We apologize for accidentally leaving out this figure, and have now added it.	2	1	Is this indeed an anti-tumor effect or a very toxic effect to the liver (including tumor).			cancers14122959_perova	0
+We thank the reviewer for this comment. In the in vitro experiments we used cell lines derived from humans. However, these cell lines cannot be implanted to rats, and so for the in vivo experiments we had to use a cell line from rats.	2	1	I read the manuscript with interest and commend the authors for the work done in the area of liver cancer treatment.			cancers14122959_perova	0
+We have introduced new several sentences in this paragraph of Materials and Methods, which define these ad-hoc rules. Lines 154-155: “mgR51C read-outs have been introduced into the classification system as PVS1_O or BP7_O codes of variable evidence strength depending on the splicing outcome [P, Sup-porting (±1 point); M, Moderate (±2); Strong (±4); Very Strong (±8)].” Lines 163-179: “we have developed some ad-hoc rules that take into consideration the coding potential of each individual transcript and its relative contribution to the overall expression to reach the appropriate PVS1_O or BP/_O evidence strength. In brief, for each complex read-out we have applied the following algorithm: (i) De-convolute mgR51C read-outs in-to individual transcripts; (ii) apply ACMG/AMP evidences to each individual transcript; (iii) produce an overall PVS1_O (or BP7_O) code strength based on the relative contribu-tion of individual transcripts/evidences to the overall expression. Thus, if pathogenic supporting transcripts contribute ≥90% to the overall expression, PVS1_O_ code is applied (if different transcripts support different pathogenic evidence strengths, the lowest strength contributing >10% to the overall expression is selected for overall evidence strength). Similarly, BP7_O_ code is applied if benign supporting transcripts contribute ≥90% to the overall expression (if different transcripts support different pathogenic evi-dence strengths, the lowest strength contributing >10% to the overall expression is selected for overall evidence strength). If neither pathogenic nor benign supporting transcripts contribute ≥90% to the overall expression, the splicing assay is considered not providing any evidence in favor, or against, pathogenicity. Recently, we have used a similar ap-proach to deal with complex PALB2/ATM minigene read-outs [20,30].” 2 -	2	1	The assay used is well established and has been used previously to evaluate spliceogenic variants in RAD51C and other hereditary breast/ovarian cancer-associated genes.			cancers14122960_makarova	0
+The five RAD51 paralogs are known to be required for homologous recombination and maintenance of genomic stability. Indeed, RAD51C interacts with RAD51B, RAD51D, XRCC2 and XRCC3 in two different complexes that play a role in homologous recombination. Miller et al studied the interaction between RAD51B and D (and also XRCC3) with deletion mutants. These authors found that Rad51C1-285 (includes β-strands 1-5) or Rad51C285-376 (includes β-strands 6-9) did not bind RAD51B. So, a complete beta-sheet is important in maintaining the overall fold of the protein. Moreover, the missense variant p.Arg312Trp (ß-strand 6) has been shown to impair RAD51C function (Gayarre et al 2017). Both studies indicate that this protein region is essential for RAD51C function so that transcripts lacking any of the β-strands, such as ▼(E6q4)-a, ▼(E6q4)-b, Δ(E7), Δ(E8) or the in-frame isoform Δ(E5), is probably deleterious. - We have modified this part, adding several sentences to clarify it. Lines 270-277: “ The integrity of the β-sheet is important in maintaining the overall fold of the RAD51C protein and the interaction with RAD51B, so that alterations of any ß-strand of RAD51C should be considered deleterious [33]. Further, structural features (the order of the ß--strands in space is not the same as their order in sequence) predict that proteins lacking any single b-strand would fail to form the ß--sheet resulting in a collapse of the protein core and misfolding of the protein [33]. Moreover, the missense variant p.Arg312Trp (ß-strand 6) has been shown to impair RAD51C function [34]. Altogether these data,…” 3-	2	1	It appears to be more appropriate as results and maybe some information could be in the introduction.			cancers14122960_makarova	0
+We have added all the transcript names and their contribution in Table 2 (Column PVS1_O/BP7_O mgR51C_ex2-8).	2	1	The lack of these protein structures are used as evidence to support pathogenicity.			cancers14122960_makarova	0
+We have included cross-references to Table 1 to facilitate understanding of the manuscript.	2	1	Using the minigene assay the authors were able to demonstrate that all 20 variants altered splicing of the minigene compared to wildtype constructs.			cancers14122960_makarova	0
+Very important comment. As the reviewer indicates, it would be essential to define the threshold of RAD51C expression from which it keeps its tumor suppressor activity. Unfortunately, it is not known by now but this finding would provide critical information to determine the pathogenicity of leaky spliceogenic variants.	2	1	Author Response Thank you very much for the positive comments 1-			cancers14122960_makarova	0
+So, we have modified this paragraph to introduce this information: “Only two variants (c.404+3A>G and c.705+3A>G) displayed 26% and 21% of the mgFL-transcript, respectively. Unfortunately, it is not known the minimal amount of RAD51C expression to confer tumor suppressor haplosufficiency so, these splicing assays were not considered informative (PVS1_O_N/A).” Note that leaky variants generate complex minigene read-outs (two or more transcripts), and are therefore classified accordingly (see methods).	2	1	I only had minor suggestions or comments: Line 156 – the authors refer to “ad-hoc rules” that they have developed for consideration of the different coding transcripts associated with the same spliceogenic variant in variant interpretation and classification.			cancers14122960_makarova	0
+Acknowledge this suggestion. We have added two references of BRCA1 and BRCA2 studies.	2	1	Although a reference is provided so that the reader can look up what these ad-hoc rules are, it would also be helpful to briefly describe these in the current manuscript.			cancers14122960_makarova	0
+In this section we have tried to describe the classification approach and the rules we have followed to classify the variants. Certainly, we agree with the referee as sometimes the method is mixed with some results. So, we have moved some sentences of the last paragraph of Materials and Methods to Results. “The PM3 evidence (in trans with a pathogenic variant in a recessive disorder) did not con-tribute to the final classification. Not surprisingly (FANCO is an extremely rare FA complementation group) [37], none of the tested variants has been identified in Fanconi Anemia patients (ClinVar and Global Variome shared LOVD databases and literature search-es). Similarly, the BS2 evidence (in trans with a pathogenic variant in a healthy individual) does not contribute to the final classification of our tested variants. Finally, we have considered that some pathogenic (PS2, PM1, PM6, PP2, PP4) and benign (BP1, BP3, BP5) codes are not applicable to the classification of RAD51C variants.” 3-	2	1	describes about the analysis of RAD51C variants using mini-gene assays for splicing defect.			cancers14122960_makarova	0
+We have included explanations of these acronyms in text and Table 1 and have modified the following sentence: “Of the 19 characterized transcripts, 14 introduced premature termination codons (PTC; PTC transcripts), and of these, 10 are predicted to be degraded by the Nonsense-Mediated Decay pathway (NMD; PTC-NMD transcripts) that is considered convincing evidence of deleteriousness (Supplementary Table S3).	2	1	Lines 259-274 – This paragraph refers to evidence used in the variant interpretation of three aberrant transcripts that kept the open reading-frame (Δ(E2p3), Δ(E5) and ▼(E8p3)).			cancers14122960_makarova	0
+We have modified the sentence of the Fl-transcript: “All variants altered splicing, 18 of which produced no traces of the mgFL-transcript or almost undetectable levels (<2.4%, c.904G>A)…”	2	1	The type of work presented in this manuscript is invaluable for accurate interpretation of spliceogenic variants that may be detected in families with suspected hereditary breast/ovarian cancer.			cancers14122960_makarova	0
+Acknowledge this comment. Splicing variants are called solely based on the size difference. It is good to look into Sanger sequence of splice variants after gel elution of the band and sequencing. We have sequenced the RT-PCR products of all variant assays (indicated in Materials and Methods, section 2.5. Minigene Splicing Assays). In fact, all the *.ab1 sequence and*.fsa fragment analysis files of RT-PCR products will be freely available at http://hdl.handle.net/10261/270934; https://doi.org/10.20350/digitalCSIC/14662 upon manuscript acceptance (links indicated in the manuscript section “Data Availability Statement”). Unfortunately, Sanger sequencing only allowed us to characterize the main transcripts, while the minor ones (<10% of the overall expression) are really difficult to characterize since gel band extraction does not work properly with these small amounts, or other methods (e.g. subcloning of RT-PCR products into a PCR-vector, or RNAseq of minigene outcomes) are laborious and not cost-effective. Anyway, we have also been using Fluorescent fragment analysis for transcript characterization in our previous studies (since Acedo et al, 2012). We have shown that this technique is highly sensitive, accurate and shows high resolution. For example, in Figure 1c, transcripts with minimal size differences (1-3 nt) are well-discriminated. So, for minor-rare transcripts is a good option (not perfect, we agree with the reviewer) to annotate them. - To clarify it, we have modified this part: Lines 132-134: “RT-PCR products were sequenced by Macrogen (Madrid, Spain), which allowed the characterization of the main variant-induced transcripts. Minor transcripts were annotated according to fluorescent fragment electrophoresis size data (see below).”	2	1	Author Response - Thank you very much for the positive comments and the revision of our manuscript.			cancers14122960_makarova	0
+We would like to thank reviewer #1 for his/her time and effort to review our manuscript. Please find our point-by-point responses below.	2	1	Conclusions: NR2F1 expression in breast cancer is associated with tumor dormancy traits and it is predominantly ex-pressed in CAFs in the tumor microenvironment.			cancers14122962_makarova	0
+We totally agree with the reviewer that it is puzzling how NR2F1 expressed in CAFs of the primary breast cancer contribute to the dormancy of DTCs, thus the addition of a discussion on this point in more detail will strengthen this manuscript. Based on our results, NR2F1 expression in primary bulk tumor is associated with several pathways related to dormancy, and NR2F1 is most predominantly expressed in CAFs in the tumor microenvironment. However, we did not prove the underlying mechanism through which CAF-expressed NR2F1 regulates dormancy, and we do not intend to claim a causal relationship. Single-cell sequence data of metastatic tumor cohorts will allow us to investigate whether the expression of NR2F1 in CAFs in the metastatic TME is related to the dormancy of DTCs. We added the following sentences to the discussion section.	2	1	To this end, we believe that NR2F1 expression in the bulk tumor does not reflect the expression in the cancer cells, thus its value as a biomarker is in doubt.			cancers14122962_makarova	0
+We agree with the reviewer that it would be interesting to see how NR2F1 levels would change with endocrine therapy. However, we do not have access to cohorts that include tumor samples before and after neoadjuvant endocrine therapy at this point. What we do have access to regarding endocrine therapy is a cohort comparing responders and non- responders to neoadjuvant endocrine therapy (GSE145325). We found that NR2F1 expression between responders and non-responders to endocrine therapy was not different. We added this to the results section as follows.	2	1	Previous studies mentioned that NR2F1 was highly expressed in DTCs [67].			cancers14122962_makarova	0
+We agree with the reviewer that breast cancer is a heterogenous disease and we should show subtype-specific data. We analyzed survival outcomes and the cell fraction in TME by each immunohistological subtype. We did not observe any validated difference in survival outcomes. Cell fractionation of immune cells and stromal cells showed almost similar trends for the scores such as intratumor heterogeneity, HRD, mutation rate, and neoantigens across all subtypes. The results for each immunohistological subtype of single-cell Cohort 2 are shown in Supplementary Figure 7, and each subtype showed the same trend. We have revised the results section and added supplementary data as follows.	2	1	(B) Boxplots showing immune and stromal cell fractions between NR2F1 high and low group in HER2 positive breast cancer of three large cohorts.			cancers14122962_makarova	0
+We thank the reviewer for reading our manuscript closely and pointing out our oversight. As you indicated, we did not find any significance difference in 2D in Figure 1A and have corrected the results section as follows.	2	1	It was also of interest to compare the expression of NR2F1 between primary and metastatic breast cancer, but there was no difference observed in the NR2F1 expression between both groups (Figure 3E).			cancers14122962_makarova	0
+We are grateful to the reviewer for her/his time and effort in reviewing our paper, as well as for pointing out issues to improve the paper.	2	1	Man-Whitney U test was used to compare the two groups and p-values are shown in bold for significant results (p < 0.05).			cancers14122962_makarova	0
+We completely agree with the reviewer that it will be informative to demonstrate the NR2F1 expression in both primary and metastatic breast cancer and the possible correlation of NR2F1 expression with metastasis. In Figure 3C, we present NR2F1 expression in primary breast cancer with and without distant metastases. NR2F1 expression was not increased in the group with later recurrence in four cohorts in this analysis. We also present NR2F1 expression between primary and metastatic breast cancer in Figure 3E with no significant difference. We did not find a clear association of NR2F1 with distant metastasis in this study. On the other hand, NR2F1 expression was higher in primary breast tumors with lymph node metastasis in all four cohorts, as shown in Figure 3B, suggesting an association between NR2F1 and lymph node metastasis. Given these results, we revised the results section as follows.	2	1	The novelty of our study is that it is CAFs, not cancer cells, that are the dominant source of NR2F1 expression in the bulk tumor.			cancers14122962_makarova	0
+We completely agree with the reviewer that NR2F1 expression in cancer cells and CAFs should be investigated not only in primary but also in metastatic breast cancer using single-cell sequence in order to prove that NR2F1 expression in CAFs affects late recurrence. The main finding of this study is that NR2F1 is predominantly expressed in CAFs rather than in all other cell types in the TME, and we do not intend to claim that CAF-expressed NR2F1 regulates dormancy. Further, we do not have access to single-cell sequence cohorts of metastatic breast cancer tumors, but it is of our interest, and this will be our future direction. In response to the reviewer, we added the following sentences in the discussion section.	2	1	Cancer mutation rates, immune responses, and immune cell infiltrations were lower in high NR2F1 tumors whereas the infiltration of stromal cells including cancer-associated fibroblasts (CAFs) was higher.			cancers14122962_makarova	0
+We thank the reviewer for closely reading our manuscript and pointing out our oversight. We have corrected the results section as answered in Responce1.	2	1	HER2- positive subtype was determined by immunohistochemistry.			cancers14122962_makarova	0
+We totally agree with the reviewer that it is puzzling how NR2F1 expressed in CAFs of the primary breast cancer contribute to the dormancy of DTCs, thus the addition of a discussion on this point in more detail will strengthen this manuscript. Based on our results, NR2F1 expression in primary bulk tumor is associated with several pathways related to dormancy, and NR2F1 is most predominantly expressed in CAFs in the tumor microenvironment. However, we did not prove the underlying mechanism through which CAF-expressed NR2F1 regulates dormancy, and we do not intend to claim a causal relationship. Single-cell sequence data of metastatic tumor cohorts will allow us to investigate whether the expression of NR2F1 in CAFs in the metastatic TME is related to the dormancy of DTCs. We added the following sentences to the discussion section.	2	1	(B) The violin plot shows NR2F1 expression in inflammatory CAFs (iCAFs) and myofibroblasts (myCAFs) by immunohistological subtypes in single-cell Cohort 2.			cancers14122962_perova	0
+We agree with the reviewer that it would be interesting to see how NR2F1 levels would change with endocrine therapy. However, we do not have access to cohorts that include tumor samples before and after neoadjuvant endocrine therapy at this point. What we do have access to regarding endocrine therapy is a cohort comparing responders and non- responders to neoadjuvant endocrine therapy (GSE145325). We found that NR2F1 expression between responders and non-responders to endocrine therapy was not different. We added this to the results section as follows.	2	1	Cancer mutation rates, immune responses, and immune cell infiltrations were lower in high NR2F1 tumors whereas the infiltration of stromal cells including cancer-associated fibroblasts (CAFs) was higher.			cancers14122962_perova	0
+We agree with the reviewer that breast cancer is a heterogenous disease and we should show subtype-specific data. We analyzed survival outcomes and the cell fraction in TME by each immunohistological subtype. We did not observe any validated difference in survival outcomes. Cell fractionation of immune cells and stromal cells showed almost similar trends for the scores such as intratumor heterogeneity, HRD, mutation rate, and neoantigens across all subtypes. The results for each immunohistological subtype of single-cell Cohort 2 are shown in Supplementary Figure 7, and each subtype showed the same trend. We have revised the results section and added supplementary data as follows.	2	1	Aguirre-Ghiso group recently demonstrated that NR2F1 agonist treatment induced cancer cell dormancy [21], which raised an expectation that NR2F1 expression in primary breast cancer may have the possibility to be a biomarker.			cancers14122962_perova	0
+We thank the reviewer for reading our manuscript closely and pointing out our oversight. As you indicated, we did not find any significance difference in 2D in Figure 1A and have corrected the results section as follows.	2	1	Comment 3: Line 170, “Figure 1E” should be “Figure 1D”.			cancers14122962_perova	0
+We completely agree with the reviewer that it will be informative to demonstrate the NR2F1 expression in both primary and metastatic breast cancer and the possible correlation of NR2F1 expression with metastasis. In Figure 3C, we present NR2F1 expression in primary breast cancer with and without distant metastases. NR2F1 expression was not increased in the group with later recurrence in four cohorts in this analysis. We also present NR2F1 expression between primary and metastatic breast cancer in Figure 3E with no significant difference. We did not find a clear association of NR2F1 with distant metastasis in this study. On the other hand, NR2F1 expression was higher in primary breast tumors with lymph node metastasis in all four cohorts, as shown in Figure 3B, suggesting an association between NR2F1 and lymph node metastasis. Given these results, we revised the results section as follows.	2	1	No significant difference was found in the NR2F1 expression with and without distant metastasis in 5 independent primary breast cancer cohorts (Figure 3B).			cancers14122962_perova	0
+We completely agree with the reviewer that NR2F1 expression in cancer cells and CAFs should be investigated not only in primary but also in metastatic breast cancer using single-cell sequence in order to prove that NR2F1 expression in CAFs affects late recurrence. The main finding of this study is that NR2F1 is predominantly expressed in CAFs rather than in all other cell types in the TME, and we do not intend to claim that CAF-expressed NR2F1 regulates dormancy. Further, we do not have access to single-cell sequence cohorts of metastatic breast cancer tumors, but it is of our interest, and this will be our future direction. In response to the reviewer, we added the following sentences in the discussion section.	2	1	Boxplots showing NR2F1 expression in responder and non-responder to neoadjuvant endocrine therapy in GSE145325.			cancers14122962_perova	0
+We thank the reviewer for closely reading our manuscript and pointing out our oversight. We have corrected the results section as answered in Responce1.	2	1	Conclusions: NR2F1 expression in breast cancer is associated with tumor dormancy traits and it is predominantly ex-pressed in CAFs in the tumor microenvironment.			cancers14122962_perova	0
+According to the reviewer suggestion, FT-IR peaks shift related to chromium ion adsorption are discussed in the mechanism section and is incorporated in the revised manuscript.	2	1	Comment 3: It is better to show the SEM images using the same scale.			catal12030290_makarova	0
+EDX analysis (In Figure 3) – is a conditional type of analysis of the chemical composition of the surface. In EDX, often, spectra with different atomic abundances of elements can be obtained even from the same sample.	2	1	Author Response We appreciate the efforts of the reviewers for their detailed and insightful comments, which have helped us to improve the quality of our manuscript.			catal12030290_makarova	0
+The sulphuric acid disintegrated the leaves. Most of the substances in the leaves are reduced to carbon after two hours. Charring takes place by adding sulphuric acid and by the action of heat, charring removes hydrogen and oxygen from the solid, so that the remaining char is composed primarily of carbon. It also helps to remove the moist content in the leaves. The pollution problem may not be takes place.	2	1	First of all, authors have to attach a EDX mapping images before/after sorption of chromium ions.			catal12030290_makarova	0
+Chromium is a potentially toxic metal occurring in water and groundwater as a result of natural and anthropogenic sources. The prepared adsorbent is well suitable in the real condition. Since the prepared adsorbent (Al-GNSC) is successfully reduces the chromium (VI) ion form the real groundwater samples.	2	1	Author Response We appreciate the efforts of the reviewers for their detailed and insightful comments, which have helped us to improve the quality of our manuscript.			catal12030290_makarova	0
+Based on the reviewer suggestion, regeneration studies were performed briefly. Four different desorption agents such as tap water, 0.1M HCl, 0.1M H2SO4 and 0.1M NaOH were utilized to remove the adsorbed chromium ions from the Al-GNSC adsorbent. From this various desorption agents it was identified that 0.1M NaOH was more effective. Hence, the reuse of Al-GNSC from Cr(VI)-loaded material was studied for sorption and desorption cycles using sodium hydroxide as a regenerated agent. Relevant references are quoted in the revised manuscript. Reference: Sujitha Ravulapalli and Ravindhranath Kunta reported that sodium hydroxide was used as a regenerating agent for the sorption and desorption of Cr(VI) by activated carbon derived from Lantana camara plant. [Enhanced removal of chromium (VI) from wastewater using active carbon derived from Lantana camara plant as adsorbent. Water Sci Technol (2018) 78 (6): 1377–1389. https://doi.org/10.2166/wst.2018.413] Reference: M.A. Tandal and B.N.OZA. reported that Sodium hydroxide as a regenerating agent for the sorption and desorption of Cr(VI) by Granular activated carbon. [Adsorption and regeneration studies for the removal of Chromium (VI) from the waste water of electroplating industry using Granular activated carbon. Asian Journal of chemistry. Vol. 17, No.4 (2005), 2524-2530] Page 2 of 2 Comment 2:	2	1	Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+We acknowledge the reviewer’s opinion. For the preparation of 1kg of Al-GNSC adsorbent, approximately 150 to 200 mL of hydrochloric acid was added in order to blend the aluminum in to the ground nut shell carbon. Thank you for your valuable comment.	2	1	Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+According to the reviewer suggestion, the expression of separation factor is mentioned in the revised manuscript. 𝑅𝐿= 1 1+𝑏𝐶𝑖, where “Ci” is the initial concentration of Cr(VI) and “b” is the Langmuir constant.	2	1	Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+However, we regret that we were not able to investigate the BET analysis due to pandemic situation, which could definitely give us additional information.	2	1	Author Response We appreciate the efforts of the reviewers for their detailed and insightful comments, which have helped us to improve the quality of our manuscript.			catal12030290_makarova	0
+According to the reviewer suggestion, the complete terms of all abbreviations are mentioned before the first use in the revised manuscript.	2	1	Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+As suggested, the novelty of the research work is explained in the introduction part of the revised manuscript, and the Page 2 of 4 obtained results are compared with the recent literature.	2	1	The authors are very thankful to the Reviewer for their valuable suggestions for the improvement of the manuscript.			catal12030290_makarova	0
+According to the reviewer suggestion, the SEM images with same scale are provided in the revised manuscript.	2	1	Author Response We appreciate the efforts of the reviewers for their detailed and insightful comments, which have helped us to improve the quality of our manuscript.			catal12030290_makarova	0
+We hope the reviewer understand the experimental deficiencies at the stage of the present experiments. We deeply appreciate the comment raised by the reviewer. Thank you very much. However, we regret that we were not able to investigate the XPS analysis due to pandemic situation, which could definitely give us additional information about the elemental confirmation.	2	1	A point-by-point response to the reviewer-1 comments is appended below for your convenience.			catal12030290_makarova	0
+According to the reviewer suggestion, XRD study has been performed and incorporated in the revised manuscript.	2	1	All the modifications are shown in yellow color in the revised manuscript.			catal12030290_makarova	0
+According to the reviewer suggestion, kinetic study has been performed and incorporated in the revised manuscript.	2	1	A point-by-point response to the reviewer-1 comments is appended below for your convenience.			catal12030290_makarova	0
+According to the reviewer suggestion, the conditions for testing adsorbents of Cr(VI) is incorporated in the Table and is mentioned in the revised manuscript.	2	1	Therefore, the authors are recommended to add the missing information (i.e.			catal12030290_makarova	0
+According to the reviewer suggestion the adsorption capacity of groundnut shell activated carbon (Qe= 7.4 mg/g) is mentioned in the revised manuscript.	2	1	All the modifications are shown in yellow color in the revised manuscript.			catal12030290_makarova	0
+Thank you for your valuable suggestion. According to the reviewer suggestion the conclusion section is elaborated with specific conclusions in the revised manuscript.	2	1	The XPS method is much more sensitive and more accurately determines changes in the chemical composition of samples.			catal12030290_makarova	0
+We acknowledge the reviewer’s opinion. As suggested by the reviewer, we checked the manuscript carefully and enlisted a professional English language service to eliminate the spelling mistakes and grammatical errors. Thank you for your valuable suggestion for strengthening the quality of the manuscript.	2	1	Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+Indeed, the liposomes are prepared by a conventional method. However, here and in our previous work [1] we use the term liposomal nanotraps to reflect the functional aspect of the liposomal action. In this context, the liposomes act as traps for bacterial toxins.	2	1	Therefore, no new information is provided with these experiments.			cells11010166_perova	0
+We verified the total protein content of the supernatants by Coomassie blue staining, which was similar for all strains (Figure. 1, for referee inspection only). The experiments were performed using different bacterial supernatant batches and results were remarkably consistent. All the supernatants were collected at the exact same bacterial culture’s optical density to harvest the bacteria in a comparable state between batches and strains. In our previous work we have shown that cholesterol-dependent cytolysins (SLO, PLY) displayed different kinetics and dynamics of their hemolytic activity compared to SLS. As a result, the total hemolytic activities of individual streptococcal supernatants were not represented by a simple sum of activities (concentrations) of their individual toxins but displayed more complex time- and amount-dependent behavior: e.g. – the activity of PLY/SLO was prevalent at the initial times of incubation and at relatively high amounts (volume) of supernatants, whereas SLS activity fully developed only after initial lag period but was prevalent when relatively low amounts of supernatants were used in the assays. The relative quantifications of toxins between streptococcal species and strains were performed in a previous publication [1] and are referred to throughout the manuscript. We added a paragraph in the result section (line 215-222) that summarizes those points. Moreover, in our current experiments we aim not only at the neutralization of the whole hemolytic secretome of streptococcus (as described in [1]) but also include (putative) cytotoxic/cytostatic activities that might be carried out either by hemolysins (SLO/SLS) or other not yet identified toxins that display cytotoxic/cytostatic but no hemolytic activities. For these reasons, in our current experiments, we use specific cytotoxic/cytostatic activities of total supernatants derived from a toxicity assay displayed in (Figure. 1), instead of concentrations of individual (partly unknown) toxins. However, we agree with the reviewers that using volume units is confusing. We therefore, edited our manuscript to display the lethal dose (LD%) interpolated from results shown in Figure 1 instead. Using LD% units accounts for the batch variability.	2	1	However, we agree with the reviewers that using volume units is confusing.			cells11010166_perova	0
+While LDH-release or MTT assay are frequently used techniques and might be preferred by some investigators, other approaches assessing cell viability might be more popular by others, dependent on particular experimental settings of a particular study. Figure 2 (for referee inspection only) demonstrates that Alamar blue cell viability assay and Trypan blue live/dead quantification provide results that are identical to those obtained in the cell proliferation protocol used in our study. We believe that the latter protocol is the most suitable experimental approach for our study since it allows distinguishing between cell lysis, cytotoxisity and cytostatic effects (i.e. between cytolysins, cytotoxins and cytostatic agents) in a single experiment. The techniques and algorithms used by the CellDrop relies on accurate and unbiased measurements, akin to previously published techniques[2,3].	2	1	The techniques and algorithms used by the CellDrop relies on accurate and unbiased measurements, akin to previously published techniques[2,3].			cells11010166_perova	0
+The focus of our study is to highlight that successful protection against the whole palette of streptococcal toxins can be achieved by using liposomal nanotraps and to show that the liposome requirements differs between bacterial species and between different types of immune cells. We agree that the SLO neutralization is not novel and simply confirm results from previous publications by us and others. In the current work we do not intend to reveal new neutralization mechanisms either for SLO or for SLS.	2	1	However, authors did not make any attempt to explore this aspect in mechanistic detail.			cells11010166_perova	0
+Streptolysin S (SLS) is a small, non-immunogenic, peptide. This means that no commercial antibody against it is available and the peptide is too small for reliable mass spectrometry detection. It is heavily post-translationally modified and is the product of a complex operon. Its exact mode-of-action is still not yet fully clarified. In a previous publication, we were able to show that SLS is neutralized by phosphatidylcholine as well as sphingomyelin liposomes [1]. However, given the poor characterization of SLS, its unavailability from commercial providers and its extremely tedious purification, the mechanistic details of the SLS neutralization will require an extensive a project of its own and are beyond the scope of the current study.	2	1	The experiments were performed using different bacterial supernatant batches and results were remarkably consistent.			cells11010166_perova	0
+We used two controls in the protection experiments. In one control we challenged the cells with bacterial supernatant without adding liposomes, to determine that the baseline cytotoxicity is in line with the results displayed in the Figure 1. The second control consists of the immune cells without toxin or liposomes. This control represents 100% survival and allows us to normalize our survival data. We also tested the intrinsic toxicity of liposomes on their own to see if they did impede cell growth or were cytotoxic. At the concentrations used in current study the liposomes were not cytotoxic (data not shown). The following text was added to the material and method section: The data were normalized to a control incubated with PBS instead of bacterial supernatant (considered as 0% cell death). A control challenged by bacterial supernatant without liposomes was added for each assay to verify the expected cytotoxic activity 2.	2	1	In the current work we do not intend to reveal new neutralization mechanisms either for SLO or for SLS.			cells11010166_perova	0
+We selected the specific supernatant volumes used to reach a similar lethal dose to compare the protection capability and efficiency of the liposomal nanotraps. However, as the immune cell lines have different sensitivities depending on the toxin profile of the tested strain, a similar lethal dose corresponds to different supernatant volumes. We used the results displayed in Figure 1 of the manuscript to determine which volume to use. We agree that the way we presented it can lead to confusion and we replaced the supernatant volume values by LD% values. The following text was added to the material and method section: The added supernatant volume was determined based on the toxicity assay results and was used either at saturating dose (lethal dose >90, LD>90) or at non-saturating dose (LD60-90) to study minor toxin activities. We agree that the distinction between saturating (LD>90) and non-saturating (LD60-90) was confusing. To clarify our process we did add clarifications throughout the results part and we added a panel in figure 2 to include results obtained after challenge of LD>90 of GGS 5804 supernatant.	2	1	Carlsen J, Cömert C, Bross P, Palmfeldt J. Optimized High-Contrast Brightfield Microscopy Application for Noninvasive Proliferation Assays of Human Cell Cultures.			cells11010166_perova	0
+All the corrections are highlighted in yellow. Corrected the legend of Figure 4 (EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT).	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+Response: The errors were corrected in text on page 09. (EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT).	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+All the corrections are highlighted in red. The “galenic” was mentioned in the article due to physician Claudio Galeno, which created the first creams formulations. Therefore we decided to change galenic to cosmetic.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+Inserted parenthesis at line 132 (Tpeak = 48.17 °C).	2	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_makarova	0
+Corrected at line 160 “Figureure” to “Figure”.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+This abbreviation is widely use in Thermal analysis. We specified in the text (the extrapolated peak onset temperature (TonsetDSC).	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+We removed the units of temperature from the Table.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+We changed “donate” to provide and the other word donor to provider.	2	1	Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			cosmetics2030248_makarova	0
+We changed this sentence.	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+The sample´s name corrected (EM/BL) at line 189.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+We integrated all the problems and correct he errors.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+We described these two points more clearly. The thermal analysis technique applies high temperatures in the samples due to this thermal stability of the emulsion associated with phytic acid decreased.	4	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_makarova	0
+All the corrections are highlighted in green. We expanded the discussion of the results on pages 7, 10 and the conclusion on page 11 (item 4).	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+As mentioned in this paper, there are few studies related to the phytic acid in cosmetic products. However we added more references regarding about phytic acid antioxidant activity. References 24, 25 and 26.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+We mentioned in the text more information about incorporation of phytic acid into emulsion on Thermal Analysis results, DPPH results and conclusion.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+Regarding with this queries we improved the discussion in the results on page 7. But we need to study more about.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+All the corrections are highlighted in yellow. Corrected the legend of Figure 4	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_perova	0
+(EM/HPHYT/BHT, EM/HPHYT, ET/HPHYT). The errors were corrected in text on page 09.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+All the corrections are highlighted in red. The “galenic” was mentioned in the article due to physician Claudio Galeno, which created the first creams formulations. Therefore we decided to change galenic to cosmetic.	2	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+Inserted parenthesis at line 132 (Tpeak = 48.17 °C).	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+Corrected at line 160 “Figureure” to “Figure”.	2	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+This abbreviation is widely use in Thermal analysis. We specified in the text (the extrapolated peak onset temperature (TonsetDSC).	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+We removed the units of temperature from the Table.	2	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+We changed “donate” to provide and the other word donor to provider.	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+We changed this sentence.	2	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+The sample´s name corrected (EM/BL) at line 189.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+We integrated all the problems and correct he errors.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+We described these two points more clearly. The thermal analysis technique applies high temperatures in the samples due to this thermal stability of the emulsion associated with phytic acid decreased.	4	1	The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+All the corrections are highlighted in green. We expanded the discussion of the results on pages 7, 10 and the conclusion on page 11 (item 4).	2	1	All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+As mentioned in this paper, there are few studies related to the phytic acid in cosmetic products. However we added more references regarding about phytic acid antioxidant activity. References 24, 25 and 26.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_perova	0
+We mentioned in the text more information about incorporation of phytic acid into emulsion on Thermal Analysis results, DPPH results and conclusion.	2	1	However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_perova	0
+Regarding with this queries we improved the discussion in the results on page 7. But we need to study more about.	2	1	Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+Following the reviewer’s comments, we have now focused on one detailed analysis based on adequate tests and a discussion of the results. The revised version drops Descriptive statistics in old Table 2, rates of increase given in the old Table 3, correlations reported in old Table 4, the regressions in old Table 5, and pooled regressions of old Table 6. The discussion based on these Tables has consequently been omitted from the revision. The unit-root-test Tables have been moved from the Appendix to the text as Table 2, and in view of the evidence in favor of unit roots, rates of growth of GDP and government variables have been shown in Table 3 somewhat like the old Table 3. These are the most major changes.	2	1	Round 2: My recommendation is to accept the paper for publication.			economies3040150_perova	0
+The authors are aware of the Johnson et al. paper (JME 2013) and other critiques of the PWT data. However, while the LCU numbers are appropriate for single-country analysis, these are not suitable for panel-data format which is a significant part of this study. Therefore, we use the PWT numbers throughout and hope that single-country analysis with these numbers is a reasonable approximation to the scenario indicated by LCU data.	2	1	Round 2: Author Response to Reviewer 1 Please note the following changes in revised version:			economies3040150_perova	0
+The dependency variable has been removed from the analysis.	2	1	Round 2: Author Response to Reviewer 1 Please note the following changes in revised version:			economies3040150_perova	0
+Figure 1 has been redone with separate scales for logarithms of real GDP per capita and government share.	2	1	Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+The correlations of old Table 4, the static regressions of Table 5, and the discussion of endogeneity have been omitted.	2	1	This makes the unconditional correlations uninteresting.			economies3040150_perova	0
+Since our data cover a long period of nearly 50 years, we are inclined to rely on Johansen tests although the number of data points is smaller than 100. The Gregory-Hansen test is done for Japan and Korea also just for completeness, and might be of some methodological interest since it does not show cointegration even for Japan and Korea. The reviewer’s observation has been acknowledged in note 2.	2	1	This makes the unconditional correlations uninteresting.			economies3040150_perova	0
+We agree with the reviewer about the limited usefulness of MTAR in small samples and for Japan and Korea. However, it is included in the hope it would indicate whether lack of cointegration for Malaysia, Philippines, Singapore and Thailand might be due to asymmetric adjustment. The reviewer’s observation has been acknowledged in note 3.	2	1	Round 2: Author Response to Reviewer 1 Please note the following changes in revised version:			economies3040150_perova	0
+We have highlighted the alternative hypothesis in Pedroni’s tests on Page 8. As Pedroni (1999) clearly points out, the alternative hypothesis rejects the null of no-cointegration across all cross-sections in both within- and between-dimension tests.	2	1	Penn World Table Revisions and Their Impact on Growth Estimates” NBER Working Paper No.			economies3040150_perova	0
+The “provocative conclusion” reflected one author’s subjective perception and was stated as such. He shares the perspective noted by Bennett McCallum’s in his “Is the spurious regression problem spurious?” (Economics Letters, 2010). However, the segment cited by the reviewer has been deleted since it is a more general proposition and is not necessary for the limited analysis reported in our paper.	2	1	Penn World Table Revisions and Their Impact on Growth Estimates” NBER Working Paper No.			economies3040150_perova	0
+We completely share the reviewer’s view that despite Biehl’s PF/FP essay, almost everyone follows the standard approach, and we do the same. Our observation is just a restatement of the proposition that our approach is traditional.	2	1	More generally, we venture the highly subjective view that despite the immense mathematical sophistication of cointegration tests and their application in perhaps thousands of studies, it is not obvious how much additional substantive insight has been gained in the wide variety of contexts in which these tests have been used.”			economies3040150_perova	0
+(a) the plots in Figure 1 have been rescaled and now there are cross-overs in the plot for Japan, (b) years have been shown on the horizontal axis in Figure 1, (c) a sizable addition has been made toward the end of Section 3 to address the reviewer’s observations about possible reasons for lack of cointegration in most countries, and (d) a few minor editorial alterations have been made.	4	1	Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+Several new references, including those kindly mentioned by the reviewer, have been added, and there is a concise description of these in note 1.	2	1	Round 2: My recommendation is to accept the paper for publication.			economies3040150_perova	0
+Several formats in which Wagner’s hypothesis has been formulated and tested have been listed on page 5 of the revision. That paragraph includes six different approaches.	2	1	Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			economies3040150_perova	0
+Real GDP per capita is a fairly standard proxy for the “level of development” in the context of Wagner’s hypothesis. It might not be appropriate to use nominal GDP per capita. For the government share, it seems better to take the current-price ratio which indicates what fraction of the current output is used for government activities. The deflators for GDP and government spending are different and “real” government-share may not be a good indicator of the resource-use by the government during the current year.	2	1	Unit roots and the demand for cigarettes in Turkey: pitfalls and possibilities.			economies3040150_perova	0
+The old Table 2 has been deleted. Those descriptive statistics were for the entire period. The growth rates in the new Table 3 are also for the entire period, and that has been indicated.	2	1	Round 1: The paper analyzes the validity of Wagner’s law in six East Asian countries during the period 1960- 2008.			economies3040150_perova	0
+The redundant graphs for Philippines have been deleted.	2	1	Round 1: The paper analyzes the validity of Wagner’s law in six East Asian countries during the period 1960- 2008.			economies3040150_perova	0
+The dependency-rate variable has been dropped.	2	1	Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+The old Table 4 has been deleted along with the remark about p-values being taken from SAS.	2	1	While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			economies3040150_perova	0
+We better specify the required items on agricultural labor productivity in the text. Sorry for the unclearness. The index is actually the measure of agricultural labor productivity, which is calculated by the agricultural output and labor. We now clarify this part with another way of expression.	2	1	Despite that, the authors must improve the paper in order to get a better piece of work.			economies4030013_makarova	0
+The Reviewer raises an interesting point that we overlooked previously. We take the starting year as 2000 is mainly because of the entry of WTO, which influences the pattern of agricultural production in China. We take the reviewer’s advice, to extend the time period until 2013, which is the latest available data on prefectural level.	2	1	We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.			economies4030013_makarova	0
+We now correct all the issues in citations and tables and figures in the text. Figure 3 (now is Figure 4), the vertical axis indicates the growth rate of agricultural labor productivity; we now put it in the note. In Appendix A1, the “absolute value” means real value; we now change it to the better expression. In Appendix A2, they are measured in real value at constant price; we now better specify it in the text.	2	1	In general, we amended the paper in accordance with the replies below.			economies4030013_makarova	0
+We would like to thank the reviewer’s scrutiny. We now specify this better in the text	2	1	Hence, all the prefectures within each province are deflated by the same index.			economies4030013_makarova	0
+The agricultural output is deflated with the base year 2000. There is no price index at the prefectural level. For this reason we use the provincial price indices to deflate the nominal series. Hence, all the prefectures within each province are deflated by the same index. Although suboptimal, this helps to preserve the comparability in the two estimations. The reviewer rises a very useful point that we initially ignored. We now better specify the required items on agricultural labor productivity in the text.	2	1	Literature discussion: Most of the literature cited in the Introduction section and Discussion section are published five years ago.			economies4030013_makarova	0
+The reviewer is right. The first sector is a macro conception of agriculture. It indeed includes fishery. However, the data of output and employment are both collected on this level - the first sector. Hence, we assume the quotient of the two variables is a proxy to agriculture labor productivity. Meanwhile, they are the only available data on the prefectural level to calculate agricultural labor productivity. Therefore, we think this calculation is still helpful.	2	1	Labor productivity needs to be measured using real value of agricultural output.			economies4030013_makarova	0
+The Reviewer raises a very valuable point. We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.	2	1	Labor productivity needs to be measured using real value of agricultural output.			economies4030013_makarova	0
+We now specify this better in the text.	2	1	The agricultural output is in current or constant prices?			economies4030013_makarova	0
+The Reviewer raises a valuable point which we overlooked previously. In the section 3.3, we improve the understanding of the growth pattern of Chinese agricultural labor productivity by adding the estimations of beta and sigma convergence. The results of convergence estimation also echo our analysis of LGCM.	2	1	Despite that, the authors must improve the paper in order to get a better piece of work.			economies4030013_makarova	0
+Yes, the Reviewer is correct. Since we add three years into the dataset (following the suggestion from the other reviewer), the table context changes. We now improve this issue in the text.	2	1	One of the most interesting issues of the paper is the political change in 2004.			economies4030013_makarova	0
+We improve this part in the text. The recent movements of the government in this agricultural reform are added. Since we add the data of recent three years, another break is observed in the curve. So we also introduce the macro policy of stimulus package.	2	1	The authors may want to have some brief discussion on more recent literature regarding China’s productivity estimates and policy related issues.			economies4030013_makarova	0
+We would like to thank the Reviewer to recommend us to develop this part. The Reviewer raises a very interesting point. However, the prefectural-level data in terms of farmland are only reported until 2007 in the China City Statistical Yearbook.	2	1	This could have strong implications in one case or another, especially in the first case.			economies4030013_makarova	0
+We better specify the required items on agricultural labor productivity in the text. Sorry for the unclearness. The index is actually the measure of agricultural labor productivity, which is calculated by the agricultural output and labor. We now clarify this part with another way of expression.	2	1	The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			economies4030013_perova	0
+The Reviewer raises an interesting point that we overlooked previously. We take the starting year as 2000 is mainly because of the entry of WTO, which influences the pattern of agricultural production in China. We take the reviewer’s advice, to extend the time period until 2013, which is the latest available data on prefectural level.	2	1	Labor productivity needs to be measured using real value of agricultural output.			economies4030013_perova	0
+We now correct all the issues in citations and tables and figures in the text. Figure 3 (now is Figure 4), the vertical axis indicates the growth rate of agricultural labor productivity; we now put it in the note. In Appendix A1, the “absolute value” means real value; we now change it to the better expression. In Appendix A2, they are measured in real value at constant price; we now better specify it in the text.	2	1	Otherwise, it may reflect the changes in prices instead of productivity.			economies4030013_perova	0
+We would like to thank the reviewer’s scrutiny. We now specify this better in the text	2	1	The authors should also clarify if this sector included the forest products and labor.			economies4030013_perova	0
+The agricultural output is deflated with the base year 2000. There is no price index at the prefectural level. For this reason we use the provincial price indices to deflate the nominal series. Hence, all the prefectures within each province are deflated by the same index. Although suboptimal, this helps to preserve the comparability in the two estimations. The reviewer rises a very useful point that we initially ignored. We now better specify the required items on agricultural labor productivity in the text.	2	1	Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to.			economies4030013_perova	0
+The reviewer is right. The first sector is a macro conception of agriculture. It indeed includes fishery. However, the data of output and employment are both collected on this level - the first sector. Hence, we assume the quotient of the two variables is a proxy to agriculture labor productivity. Meanwhile, they are the only available data on the prefectural level to calculate agricultural labor productivity. Therefore, we think this calculation is still helpful.	2	1	Data issue: The authors indicate their data source as “…an index of panel data on agricultural output and employment in the first sector of 287 prefectures from 2000 to 2010 is collected from the China city Statistical Yearbook (2001-2011)...” It is not clear what kind of index the authors are referring to.			economies4030013_perova	0
+The Reviewer raises a very valuable point. We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.	2	1	Issues to improve the paper: The introduction is welldone, but it could be improved in some points.			economies4030013_perova	0
+We now specify this better in the text.	2	1	Table 3: Each column title needs to be more specific.			economies4030013_perova	0
+The Reviewer raises a valuable point which we overlooked previously. In the section 3.3, we improve the understanding of the growth pattern of Chinese agricultural labor productivity by adding the estimations of beta and sigma convergence. The results of convergence estimation also echo our analysis of LGCM.	2	1	The authors should also clarify if this sector included the forest products and labor.			economies4030013_perova	0
+Yes, the Reviewer is correct. Since we add three years into the dataset (following the suggestion from the other reviewer), the table context changes. We now improve this issue in the text.	2	1	If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions?			economies4030013_perova	0
+We improve this part in the text. The recent movements of the government in this agricultural reform are added. Since we add the data of recent three years, another break is observed in the curve. So we also introduce the macro policy of stimulus package.	2	1	Author Response We would like to thank the Reviewers for a very careful reading of the paper and a number of interesting, though challenging, suggestions and comments.			economies4030013_perova	0
+We would like to thank the Reviewer to recommend us to develop this part. The Reviewer raises a very interesting point. However, the prefectural-level data in terms of farmland are only reported until 2007 in the China City Statistical Yearbook.	2	1	Thanks to his/her remarks, the paper has improved under many important aspects.			economies4030013_perova	0
+According to reviewer’s comment, we added Equation (14), and Tables 3 and 4 on the number of additions for the VVC inverse transforms.	2	1	It is better that they replace them with other keywords to increase the reach of the manuscript.			electronics11050760_makarova	0
+According to the reviewer’s comment, we modified and added some keywords as follows: We replaced “Keywords: VVC; HEVC; video coding; transform; computational complexity” With “Keywords: VVC (Versatile Video Coding), HEVC (High Efficiency Video Coding, Linear Inverse Transform, computational complexity, DCT (Discrete Cosine Transform), Discrete Sine Transform (DST), BD-rate”	2	1	We included the following sentence and Equation (14) on page 4: “For an (n×m) transform block which has N non-zero coefficients, the total number of additions in the proposed inverse transform using linearity is computed in Equation (14), even though Equation (14) is not used in the proposed method: We included the following Table 3 and 4 and sentences on page 6: Table 3.			electronics11050760_makarova	0
+According to the reviewer's comments, we updated the last paragraph of Section 1 as follows: We replaced “Section 2 presents a proposed inverse transform using linearity to reduce the computational complexity.” With “In Section 2.1, we introduce the transforms used in VVC. And we propose a fast inverse transform using linearity to reduce the computational complexity in Section 2.2.” Comment) Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	2	1	References List: The list of references is recent, and all references are related to the research topic but it is not sufficient for this study.			electronics11050760_makarova	0
+According to the reviewer’s comment, we divided Section 2 as follows: 2. VVC Transforms and Proposed Method 2.1. Introduction to DCT-II, DST-VII, and DCT-VIII 2.2. Propose Fast Inverse Transform Using Linearity Comment) The authors should add a section where they discuss comparing their results with those of existing research.	2	1	Zhao, X.; Kim, S,-H,; Zhao, Y.; Eglimez, H.; Koo, M.; Liu, S.; Lainema, J.; Karczewicz, M. Transform Coding in the VVC Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol.			electronics11050760_makarova	0
+According to the reviewer’s comment, we included the following sentence in the end of 3. Experimental Results section on page 11: ~~ “Fast encoding methods only in the encoder side were proposed to reduce the encoding complexity of VVC but all the fast encoding methods increased the BD-rates [29][30][31] in terms of the bit-rate reduction so that the proposed inverse transform using linearity in the decoder side differs from those approaches in that it keeps the BD-rate in VVC while reducing the decoding complexity. If the proposed inverse transform is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. Therefore, the proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.” ~~ And three new references [29],[30], and [31] in Reference section were included.	2	1	It can be easily computed by Equation (14) that the number of additions of the proposed method is smaller for all block sizes than those in VTM-8.2.”			electronics11050760_makarova	0
+According to the reviewer’s comment, we added the future directions in the conclusion section as follows: “The proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.”	2	1	The authors should add the main contributions briefly at the end of the introduction.			electronics11050760_makarova	0
+High Efficient Video Coding (HEVC), Standard ITU-T Recommendation H.265 and ISO/IEC 23008-2, Apr. 2013. Sullivan, G. J.; Ohm, J.; Han, W.; Wiegand, T. Overview of the High Efficiency Video Coding (HEVC) Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2012, vol. 22, pp. 1649-1668, doi: 10.1109/TCSVT.2012.2221191. Bross, B.; Chen, J.; Liu, S.; Wang, Y.-K. Versatile Video Coding (Draft 10). In Proceedings of the 19th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Teleconference (Online), 22 June-1 July 2020; 6. Bross, B.; Wang, Y.-K; Ye, Y,; Liu, S.; Sullivan, G. J.; Ohm, J. Overview of the Versatile Video Coding (VVC) Standard and its Applications, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol. 31, pp. 3736-3764, doi: 10.1109/TCSVT.2021.3101953 7. Karhunen, K. Über Lineare Methoden in der Wahrscheinlichkeitsrechnung, Soumalainen Tiedeakatemia, 1947; pp. 1-79.	2	1	Inverse Transform Using Linearity Hyeonju Song*, and Yung-Lyul Lee* *Digital Media System Laboratory, Department of Computer Engineering, Sejong University, Seoul, Korea.			electronics11050760_makarova	0
+Thank you for pointing out our mistake. According to the reviewer’s comment, we completely updated the list of references as follows: 1. Advanced Video Coding (AVC), Standard ITU-T Recommendation H.264 and ISO/IEC 14496-10, May 2003. Wiegand, T.; Sullivan, G. J.; Bjontegaard, G.; Luthra, A. Overview of the H.264/AVC Video Coding Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2003, vol. 13, pp. 560-576, doi: 10.1109/TCSVT.2003.815165..	2	1	Please add information on the number of additions for different transform types and sizes in original and proposed variants.			electronics11050760_makarova	0
+According to the reviewer’s comment, the revised manuscript was reviewed and substantially modified by an English native speaker.	2	1	Hung, C. -Y.; Landman, P. Compact inverse discrete cosine transform circuit for MPEG video decoding, In 1997 IEEE Workshop on Signal Processing Systems.			electronics11050760_makarova	0
+According to reviewer’s comment, we the following sentences at the end of the Introduction. “In this paper, we analyze the number of multiplications of the existing fast transform methods in the VVC standard, and we propose a new fast inverse transform using the number of non-zero coefficients based on linearity to reduce the number of multiplications.” Comment 2) Discussion Section: The authors did not respond accurately to this comment.	4	1	The results show a higher performance of the proposed computation of inverse transforms.			electronics11050760_makarova	0
+According to reviewer’s comment, we added Discussion Section. Discussion The previously proposed fast methods were mainly addressed to reduce complexity in the video encoder with the BD-rate loss. In [29], a fast intra mode decision algorithm was proposed and the result showed the encoding time savings of 51%~53% with BD-rate loss of 0.93%~1.08%. A low-complexity CTU (Coding Tree Unit) partition structure decision and fast intra mode decision were proposed in [30] and showed the average encoding time saving of 63% with the BD-rate loss of 1.93%. The fast encoders for video coding reduce only the encoder complexity but the BD-rates were always increased without decreasing the decoder complexity. However, the proposed fast inverse transform is different from the fast encoders in that it reduces the complexity in both the encoder and decoder while maintaining the BD- rate of the VVC standard. In the RA configuration, the proposed method reduces the average encoding and decoding times by approximately (4, 10) %, respectively, while maintaining average BD-rates. If the proposed inverse transform using the number of non-zero coefficients is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. This fact can be demerit. However, the proposed method can be considered in the next-generation video coding standards because it achieves decoding run-time saving, while maintaining average BD-rate. In addition to that, the proposed method is more effective in high QP value than in low QP value, because the higher the QP value is, the fewer no-zero coefficients there are.	4	1	Comment) Conclusions: Future directions should be added to the Conclusion Section.			electronics11050760_makarova	0
+We cannot find them.	4	1	Schwarz, H.; Coban, M.; Karczewicz, M.; Chuang, T.-D.; Bossen, F.; Alshin, A.; Lainema, J.; Helmrich, C.; Wiegand, T. Quantization and Entropy Coding in the Versatile Video Coding (VVC) Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol.			electronics11050760_makarova	0
+According to reviewer’s comment, we added Equation (14), and Tables 3 and 4 on the number of additions for the VVC inverse transforms.	2	1	Some search names in the reference list begin an uppercase letter for each word (such as [4], [5] ...			electronics11050760_perova	0
+According to the reviewer’s comment, we modified and added some keywords as follows: We replaced “Keywords: VVC; HEVC; video coding; transform; computational complexity” With “Keywords: VVC (Versatile Video Coding), HEVC (High Efficiency Video Coding, Linear Inverse Transform, computational complexity, DCT (Discrete Cosine Transform), Discrete Sine Transform (DST), BD-rate” Comment) Introduction: The authors should add the main contribution s briefly at the end of the introduction.	2	1	Koo, M.; Salehifar, M.; Lim, J.; Kim, S.-H. Low Frequency Non-Separable Transform (LFNST), 2019 Picture Coding Symposium (PCS), 2019, pp. 1-5, doi: 10.1109/PCS48520.2019.8954507.			electronics11050760_perova	0
+According to the reviewer's comments, we updated the last paragraph of Section 1 as follows: We replaced “Section 2 presents a proposed inverse transform using linearity to reduce the computational complexity.” With “In Section 2.1, we introduce the transforms used in VVC. And we propose a fast inverse transform using linearity to reduce the computational complexity in Section 2.2.” Comment) Methodology Section: It should be structured, sub-headings should be added to facilitate tracking and understanding.	2	1	Bjontegaard, G. Calculation of Average PSNR Differences Between Rd-Curves, Austin, TX, USA, pp.			electronics11050760_perova	0
+According to the reviewer’s comment, we divided Section 2 as follows: 2. VVC Transforms and Proposed Method 2.1. Introduction to DCT-II, DST-VII, and DCT-VIII 2.2. Propose Fast Inverse Transform Using Linearity Comment) The authors should add a section where they discuss comparing their results with those of existing research.	2	1	Comment) Figures and Tables: All figures and tables are shown before they are used in the text Answer)			electronics11050760_perova	0
+According to the reviewer’s comment, we included the following sentence in the end of 3. Experimental Results section on page 11: ~~ “Fast encoding methods only in the encoder side were proposed to reduce the encoding complexity of VVC but all the fast encoding methods increased the BD-rates [29][30][31] in terms of the bit-rate reduction so that the proposed inverse transform using linearity in the decoder side differs from those approaches in that it keeps the BD-rate in VVC while reducing the decoding complexity. If the proposed inverse transform is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method. Therefore, the proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.” ~~ And three new references [29],[30], and [31] in Reference section were included.	2	1	Koo, M.; Salehifar, M.; Lim, J.; Kim, S.-H. Low Frequency Non-Separable Transform (LFNST), 2019 Picture Coding Symposium (PCS), 2019, pp. 1-5, doi: 10.1109/PCS48520.2019.8954507.			electronics11050760_perova	0
+According to the reviewer’s comment, we added the future directions in the conclusion section as follows: “The proposed method can be considered in the next-generation video coding standards because it reduces the decoding complexity while the BD-rate is maintained.”	2	1	Authors should check the entire article to remove all extensive mistakes (grammatical and typos) and to improve English writing quality.			electronics11050760_perova	0
+Thank you for pointing out our mistake. According to the reviewer’s comment, we completely updated the list of references as follows: 1. Advanced Video Coding (AVC), Standard ITU-T Recommendation H.264 and ISO/IEC 14496-10, May 2003. 2. Wiegand, T.; Sullivan, G. J.; Bjontegaard, G.; Luthra, A. Overview of the H.264/AVC Video Coding Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2003, vol. 13, pp. 560-576, doi: 10.1109/TCSVT.2003.815165.. 3. High Efficient Video Coding (HEVC), Standard ITU-T Recommendation H.265 and ISO/IEC 23008-2, Apr. 2013. 4. Sullivan, G. J.; Ohm, J.; Han, W.; Wiegand, T. Overview of the High Efficiency Video Coding (HEVC) Standard, In IEEE Transactions on Circuits and Systems for Video Technology, 2012, vol. 22, pp. 1649-1668, doi: 10.1109/TCSVT.2012.2221191. 5. Bross, B.; Chen, J.; Liu, S.; Wang, Y.-K. Versatile Video Coding (Draft 10). In Proceedings of the 19th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Teleconference (Online), 22 June-1 July 2020; 6. Bross, B.; Wang, Y.-K; Ye, Y,; Liu, S.; Sullivan, G. J.; Ohm, J. Overview of the Versatile Video Coding (VVC) Standard and its Applications, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol. 31, pp. 3736-3764, doi: 10.1109/TCSVT.2021.3101953 7. Karhunen, K. Über Lineare Methoden in der Wahrscheinlichkeitsrechnung, Soumalainen Tiedeakatemia, 1947; pp. 1-79. 8. Ahmed, N.; Natarajan, T.; Rao, K. R. Discrete Cosine Transform, In IEEE Transactions on Computers, 1974, vol. C-23, pp. 90-93, doi: 10.1109/T-C.1974.223784. 9. Rose, K.; Heiman, A.; Dinstein, I. DCT/DST Alternate-Transform Image Coding, In IEEE Transactions on Communications, 1990, vol. 38, pp. 94-101, doi: 10.1109/26.46533. 10. Zhao, X.; Chen, J.; Karczewicz, M.; Zhang, L.; Li, X.; Chien, W. -J. Enhanced Multiple Transform for Video Coding, 2016 Data Compression Conference (DCC), 2016, pp. 73-82, doi: 10.1109/DCC.2016.9. 11. Han, J.; Saxena, A.; Melkote, V.; Rose, K. Jointly Optimized Spatial Prediction and Block Transform for Video and Image Coding, In IEEE Transactions on Image Processing, 2012, vol. 21, pp. 1874-1884, doi: 10.1109/TIP.2011.2169976. 12. Budagavi, M.; Fuldseth, A.; Bjøntegaard, G.; Sze, V.; Sadafale, M. Core Transform Design in the High Efficiency Video Coding (HEVC) Standard, In IEEE Journal of Selected Topics in Signal Processing, 2013, vol. 7, pp. 1029-1041, doi: 10.1109/JSTSP.2013.2270429. 13. Zhao, X.; Chen, J.; Karczewicz, M.; Said, A.; Seregin, V. Joint Separable and Non-Separable Transforms for Next-Generation Video Coding, In IEEE Transactions on Image Processing, 2018, vol. 27, no. 5, pp. 2514-2525, doi: 10.1109/TIP.2018.2802202. 14. Zhao, X.; Chen, J.; Said, A.; Seregin, V.; Egilmez, H. E.; Karczewicz, M. NSST: Non-separable secondary transforms for next generation video coding, 2016 Picture Coding Symposium (PCS), 2016, pp. 1-5, doi: 10.1109/PCS.2016.7906344.	2	1	30, no. 6, pp. 1668-1682 doi: 10.1109/TCSVT.2019.2904198.			electronics11050760_perova	0
+According to reviewer’s comment, we the following sentences at the end of the Introduction. “In this paper, we analyze the number of multiplications of the existing fast transform methods in the VVC standard, and we propose a new fast inverse transform using the number of non-zero coefficients based on linearity to reduce the number of multiplications.” Comment 2) Discussion Section: The authors did not respond accurately to this comment.	4	1	IEEE Transactions on Multimedia, 2022, vol. 24, pp. 400-414, doi: 10.1109/TMM.2021.3052348.			electronics11050760_perova	0
+4. According to reviewer’s comment, we added Discussion Section. Discussion The previously proposed fast methods were mainly addressed to reduce complexity in the video encoder with the BD-rate loss. In [29], a fast intra mode decision algorithm was proposed and the result showed the encoding time savings of 51%~53% with BD-rate loss of 0.93%~1.08%. A low-complexity CTU (Coding Tree Unit) partition structure decision and fast intra mode decision were proposed in [30] and showed the average encoding time saving of 63% with the BD-rate loss of 1.93%. The fast encoders for video coding reduce only the encoder complexity but the BD-rates were always increased without decreasing the decoder complexity. However, the proposed fast inverse transform is different from the fast encoders in that it reduces the complexity in both the encoder and decoder while maintaining the BD- rate of the VVC standard. In the RA configuration, the proposed method reduces the average encoding and decoding times by approximately (4, 10) %, respectively, while maintaining average BD-rates. If the proposed inverse transform using the number of non-zero coefficients is applied to the VVC standard, the inverse transform of the VVC standard should be changed to include the proposed method.	4	1	<Reviewer 2> “The authors should accurately address the below comments.” We would like to thank the reviewer for the thorough review and very helpful comments.			electronics11050760_perova	0
+We cannot find them.	4	1	Table 4 presents the number of additions computed in the VTM-8.2 source code in each (n  m) block size when the horizontal and vertical transforms are a combination of DST-VII and DCT-VIII.			electronics11050760_perova	0
+The diesel generator equal to the rating of base load is kept as backup to avoid total black out situation of the system.	2	1	Point 6: Sizing of renewable energy sources and battery systems were not properly explored.			en15031006_perova	0
+According to your suggestion, we have improved the quality of Figures as can see Figure 1 and Figure2.	2	1	Point 2: In such a study an experimental case must be presented in a specified region Response 2: Thank you for your comment.			en15031006_perova	0
+According to your suggestion, we have modified the Equations 4, 9 and 12 in the revised version of the manuscript as fellows.	2	1	Point 5: It could be better to include base load ratings in the results section (Fig: 10) Response 5:			en15031006_perova	0
+According to your suggestion the baseload is added in the result section as follows: In addition, to further validate the proposed system, the appliance schedule pattern for Monday is drawn in Figure 10 and Figure 9 represents the appliance weightage given by consumers for Monday. In addition, Figure 8 represents the power generated graph for the same day. The baseload is calculated as 1000 watts for each house to keep basic appliances turn ON uninterruptedly. As there are 5 houses in the community the total base power for the system is 5000 watts. The excess power is utilize to turn ON the schedulable appliances on Monday.	2	1	Point 7: Please discuss how consumers’ comfortability is marinated in the proposed method.			en15031006_perova	0
+As Sizing of the grid system is not the main goal of the article, but as per your suggestion, it is briefly discuss in the subsections of Mathematical modelling of the Energy sources. Both solar and wind systems are designed for output power equal to the consumer’s load whose annual load curve peak is maximum out of all the consumer’s in the community. Battery storage systems have the capability to supply uninterrupted power equal to the base load of the system up to 6 hours. And diesel generator equal to the rating of system’s base load is kept as a backup to avoid total black out situation. The designed system is not the time-varying tariff, therefore the overall system is designed to treat each consumer of the community equally. In time-varying tariff based system, the sizing of the system is the main objective as to accommodate the wealthy consumers of the community.	2	1	As the proposed system is in isolated mode, how continuity of power supply be maintained?			en15031006_perova	0
+According to your suggestion the consumer comfortability is discussed from line 144-160. In this work, a DSM algorithm is proposed by using renewable resources PV and wind turbines for those houses that are located far away from the main city.	2	1	As the proposed system is in isolated mode, how continuity of power supply be maintained?			en15031006_perova	0
+According to your suggestion, in the revised version of the manuscript, the unnecessary use of subjective pronoun is removed and overall writing of the paper is improved.	2	1	Point 3: The quality of the figures needs to be improved.			en15031006_perova	0
+Thank you for your comment. Due to the lack of real time datasets, we have used the publically available datasets for the verification of the proposed system.	2	1	Point 6: Sizing of renewable energy sources and battery systems were not properly explored.			en15031006_perova	0
+Battery storage bank is included in the system to provide the baseload power in the time when renewable sources are not suffice to meet the baseload power demand. Diesel generators equal to the rating of base load is kept as backup to avoid total black out in worse conditions. Our objective is to schedule and shift the controllable appliances by estimating hourly power generation and considering consumer demand as a variable function. The appliances are prioritized based on their need and demand to achieve optimum energy utilization. The main contribution of this paper include: A demand side management algorithm is proposed to fulfil the energy gap between generation and consumer’s demand for standalone renewable energy system. K-mean clustering is used to make clusters of the data based on two factors: probability of turning ON a specific appliance at time t and priority number given by consumer to that specific appliance. Linear integer programming is used to schedule the appliances clusters based on the available power and state of charge of the battery system. According to your suggestion, the novelty of the proposed work has been made more clear and understandable for readers in the introduction section as follows: In this work, a DSM algorithm is proposed by using renewable resources PV and wind turbines for those houses that are located far away from the main city.	2	1	Point 4: Equations 4, 9, and 12 include some corrections and need to be modified.			en15031006_perova	0
+According to your suggestion, we have modified the literature review and included some latest references i.e., [22][29][30][32]. In addition, we organized the literature review in more comprehensive way.	2	1	Point 2: In such a study an experimental case must be presented in a specified region Response 2: Thank you for your comment.			en15031006_perova	0
+According to your suggestion, we have improved the Figures as can be seen in the revised version of the manuscript.	2	1	Point 5: It could be better to include base load ratings in the results section (Fig: 10) Response 5:			en15031006_perova	0
+We have taken your advice and changed statistical methods in the manuscript.	2	1	Page 2, Lines 66 and 67: Write the microorganisms' names in italic.			fermentation8050212_makarova	0
+We have improved English language.	2	1	"Page 3, Line 124: Change ""ethyl acetate reagents"" to "" ethyl acetate reagent""."			fermentation8050212_makarova	0
+FRBE is water-soluble fermented rice bran extracts.	2	1	Page 2, Lines 66 and 67: Write the microorganisms' names in italic.			fermentation8050212_makarova	0
+The hot water extraction conditions was adopt according to previous methods (Dong et al., 2021). Dong, C.L. ; Liu, N.; Wang, Y.; Song, M.; Wang, R.F. ; Yang, Y.P. ; An, X.P. Study on Antioxidant Activity of Fermented Rice Bran Water Extract. Feed Industry 2021, 42(24), 8-13. DOI:10.13302/j.cnki.fi.2021.24.002.	2	1	This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			fermentation8050212_makarova	0
+24hpf means 24 hours post fertilization.	2	1	"Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology."			fermentation8050212_makarova	0
+AO solution is acridine orange.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?			fermentation8050212_makarova	0
+; Qi, J.W. Humates added to feed stimulate the microbial growth(Huck et al., 1991). Sodium humate added to fermentation substrate improved the growth of bacteria (Liu et al., 2014, Shi et al., 2016). The role of sodium humate is fermentation synergist. Huck, T.A., N. Porter and M.E. Bushell, 1991. Effect of humates on microbial activity. Gen. Microbiol. 137: 2321-2329. Liu, N.; An, X.P. ; Tong, B. S.; Chen, D. Y.; Yu, C.Q. Study on the effect of sodium humate on solid fermentation of lactobacillus plantarum. Chinese Journal of Animal Science 2014, 50(05),79-82. Shi, J.X. ; Qi, J.W; An, X.P. ; Liu, N.; Luo, X.G; Chen, D.Y. The influence of sodium humate on fermentation characteristics of yeast. Feed Industry, 2016, 37(04), 44-47.	2	1	The main comment addressing the article improvement is the methodology applied for statistical analysis.			fermentation8050212_makarova	0
+It is my mistake to write the water-soluble polysaccharide content of FRBE and RBE on the wrong sides. The water-soluble polysaccharide content values have been corrected.	2	1	"Page 3, Line 124: Change ""ethyl acetate reagents"" to "" ethyl acetate reagent""."			fermentation8050212_makarova	0
+We have taken your advice to change the number of decimal places.	2	1	Since I have some doubts that I would like clarified, I recommend that a major revision be warranted.			fermentation8050212_makarova	0
+Phenolic acids belongs to polyphenols. The increasing of content of water-soluble phenolic acids in FRBE may be attributed to the degradation of rice bran cell wall during fermentation.	2	1	For your suggestion we have dealt with the comments as follows: Comments to the Author			fermentation8050212_makarova	0
+Fig.3a and 3b showing different groups of LPO rate.	2	1	"Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology."			fermentation8050212_makarova	0
+The 73.85 and 77.00 values are correct.	2	1	"""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH."			fermentation8050212_makarova	0
+We have rewrited the first sentence in Conclusions. In summary, the DPPH radical radical scavenging abilities of rice bran water-soluble extracts and the content of bioactive constituents were considerably altered by mixed bacteria solid-state fermentation.	2	1	(2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”.			fermentation8050212_makarova	0
+I have changed.	2	1	In the reference list, write the names of the microorganisms in italic.			fermentation8050212_makarova	0
+The abbreviated form of some journal titles have been corrected to full form taking into account the rules of the journal.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?			fermentation8050212_makarova	0
+The statistical analysis were conducted as follows: All experiments were conducted in triplicate. The UNIVARIATE procedure of SAS was used to test the normality of the data before any further analyses were carried out. Data were subjected to ANOVA using the MIXED procedure of SAS (SAS 9.2, SAS Institute Inc., Cary, NC). Mean ± standard error (S.E.) were reported, and means were separated by LSD multiple comparisons. Differences were accepted as statistically significance at P < 0.05.	2	1	"Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology."			fermentation8050212_makarova	0
+Thanks for your comment. We have enhanced clarity and detail of the image for easy reading.	2	1	The application of the Duncan test should be justified because it is less sensitive in this kind of study.			fermentation8050212_makarova	0
+We have taken your advice and changed statistical methods in the manuscript.	2	1	For your suggestion we have dealt with the comments as follows:			fermentation8050212_perova	0
+We have improved English language.	2	1	It appears on Page 4, Line 162; Explain the role of sodium humate.			fermentation8050212_perova	0
+FRBE is water-soluble fermented rice bran extracts.	2	1	Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al.			fermentation8050212_perova	0
+The hot water extraction conditions was adopt according to previous methods (Dong et al., 2021). Dong, C.L. ; Liu, N.; Wang, Y.; Song, M.; Wang, R.F. ; Yang, Y.P. ; An, X.P. ; Qi, J.W. Study on Antioxidant Activity of Fermented Rice Bran Water Extract. Feed Industry 2021, 42(24), 8-13. DOI:10.13302/j.cnki.fi.2021.24.002.	2	1	In the reference list, write the names of the microorganisms in italic.			fermentation8050212_perova	0
+24hpf means 24 hours post fertilization.	2	1	Since I have some doubts that I would like clarified, I recommend that a major revision be warranted.			fermentation8050212_perova	0
+AO solution is acridine orange.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?			fermentation8050212_perova	0
+Humates added to feed stimulate the microbial growth(Huck et al., 1991). Sodium humate added to fermentation substrate improved the growth of bacteria (Liu et al., 2014, Shi et al., 2016). The role of sodium humate is fermentation synergist.	2	1	Standardize, taking into account the rules of the journal.			fermentation8050212_perova	0
+It is my mistake to write the water-soluble polysaccharide content of FRBE and RBE on the wrong sides. The water-soluble polysaccharide content values have been corrected.	2	1	Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			fermentation8050212_perova	0
+We have taken your advice to change the number of decimal places.	2	1	Table 1: The number of decimal places of some constituents is excessive.			fermentation8050212_perova	0
+Phenolic acids belongs to polyphenols. The increasing of content of water-soluble phenolic acids in FRBE may be attributed to the degradation of rice bran cell wall during fermentation.	2	1	"Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology."			fermentation8050212_perova	0
+Fig.3a and 3b showing different groups of LPO rate.	2	1	1703121)’ for publication in the Fermentation.			fermentation8050212_perova	0
+The 73.85 and 77.00 values are correct.	2	1	Page 3, Line 124: What do you mean “oscillated for 10 minutes”?			fermentation8050212_perova	0
+We have rewrited the first sentence in Conclusions. In summary, the DPPH radical radical scavenging abilities of rice bran water-soluble extracts and the content of bioactive constituents were considerably altered by mixed bacteria solid-state fermentation.	2	1	Why did the water-soluble phenolic acids contents increase after the fermentation?			fermentation8050212_perova	0
+The “oscillated for 10 minutes” means “shaked for 10 minutes”.	2	1	Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			fermentation8050212_perova	0
+I have changed.	2	1	Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_perova	0
+The abbreviated form of some journal titles have been corrected to full form taking into account the rules of the journal.	2	1	"Page 4, Line 133: Change ""VWD detector"" to ""Variable wavelength detector (VWD)""."			fermentation8050212_perova	0
+The statistical analysis were conducted as follows: All experiments were conducted in triplicate. The UNIVARIATE procedure of SAS was used to test the normality of the data before any further analyses were carried out. Data were subjected to ANOVA using the MIXED procedure of SAS (SAS 9.2, SAS Institute Inc., Cary, NC). Mean ± standard error (S.E.) were reported, and means were separated by LSD multiple comparisons. Differences were accepted as statistically significance at P < 0.05.	2	1	It appears for the first time on Page 1, Line 17; The hot water extraction conditions were tested before?			fermentation8050212_perova	0
+Thanks for your comment. We have enhanced clarity and detail of the image for easy reading.	2	1	It appears on Page 4, Line 162; Explain the role of sodium humate.			fermentation8050212_perova	0
+Thanks for your suggestion. In this study, texture profile analysis was investigated by a Texture Analyzer (TA.XT PLUS/50, STABLEMICVO, UK) using a P/0.5R cylindrical probe, and corresponding description has been added to the manuscript in Line 526 marked in red font.	2	1	The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_makarova	0
+The gel without the emulsion was taken as the emulsion blank gel, which was the gel formed only by gel continuous phase.	2	1	This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			gels8040212_makarova	0
+Wet gel were the emulsion gels we obtained, which had a three-dimensional network structure that can hold a large amount of water. The corresponding Line 570 in the manuscript has been revised to a more accurate expression, emulsion gels. Thanks again for your careful review.	2	1	What equipment was used for the texture profile analysis?			gels8040212_makarova	0
+Thanks for carefully checking. Whey protein isolate (WPI) has been added in Line 60-61 in the manuscript.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+Thanks for your constructive comment. Line 84-88 has been removed and it made the manuscript more logically. Thank you again for the good comment.	2	1	Author Response Point 1: This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			gels8040212_makarova	0
+The emulsions of different droplet size was prepared at WPI concentration of 1.5 wt% and this information has been added in Line 102. This concentration was selected as a suitable emulsifier concentration based on the results in Figure 1a and 1b, and more details can be found in the manuscript.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+The manuscript has been revised accordingly in Line 159-161.	2	1	The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_makarova	0
+Figure 3 has been revised accordingly.	2	1	Thanks very much for your contribution to our manuscript.			gels8040212_makarova	0
+Figure 4 has been adjusted to make the legend clear. In fact, B1, B2 and B3 were in a relatively homogeneous state, and no phase separation had occurred. The shadow in the middle part of the apparent image of the heat-induced gels was because the texture test was carried out before shooting. Unlike TGase-induced emulsion gel had very good elastic texture, the structure of heat-induced gels did not fully recover to its original state due to its soft and less elastic texture after compression.	2	1	The manuscript is presented in a well-structured manner.The references cited are relevant and current, mostly published within the last three years.The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_makarova	0
+Thanks for your constructive suggestion. Figure 7B has been revised accordingly, that is, a graph was added which displayed the apparent viscosity of all emulsion gels at a fixed shear rate of 0.25/s, showing the differences between groups very visually. Thanks again for your suggestion.	2	1	One main request would be to make it clearer what was the formulations and conditions tested.			gels8040212_makarova	0
+In order to choose appropriate emulsifier, we prepare different WPI concentration of 1.0-2.0 wt% and found the emulsion formulated with 1.5 wt% WPI had smaller size and medimum zeta potential, so it was described to the optimize condition. For clarity, we have also revised the corresponding positions in the manuscript.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+Thanks for your good suggestion. The gel continuous phase was the WPI-gelatin mixture. To more clearly describe the composition of the emulsion gel, Table 3 has been added to the manuscript. The pre-gel mixture, refered to the mixture before gelation occurred, was composed of WPI-gelatin mixture and emulsions, but due to the large number of samples prepared by the single factor experiment, only the general formulation of the emulsion gel was listed in Table 3. And the relevant statements in 4.2.2 and 4.2.3 have been revised in the manuscript.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+Thanks for your check. It was revised in the manuscript.	2	1	The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_makarova	0
+Thanks for your carefully review. The characterization of emulsions was investigated by the droplet size, polydispersity index (PDI) and zeta potential. This expression has been revised in the manuscript. Point 14: Line 500: Define PDI at first use	2	1	Line 60: define WPI at first use Line 84-88: This is a summary of what was carried out and not a discussion, please remove Figure 1: Include in Fig.1c what was the the % of WPI used Figure 2: Increase the font of the legend Line 166: English check Figure 3: rename using latin alphabet Figure 4: Increase legend as it is not visible.			gels8040212_makarova	0
+PDI was the polydispersity index and the manuscript has been revised in Line 501.	2	1	This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			gels8040212_makarova	0
+The gap was set to 1 mm. Thank you again for your careful review.	2	1	This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			gels8040212_makarova	0
+Thanks for your question. 1% strain was within the linear viscoelastic region in our pre-experiment so it was chosen for other tests.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+Thanks for your comment. The statement in 4.2.5 has been revised in the manuscript.	2	1	Figure 2 has been modified accordingly to make the legend clearer.			gels8040212_makarova	0
+Thanks for your carefully check. Line 533 has been modified which corresponding to the line of revised manuscript.	2	1	Line 60: define WPI at first use Line 84-88: This is a summary of what was carried out and not a discussion, please remove Figure 1: Include in Fig.1c what was the the % of WPI used Figure 2: Increase the font of the legend Line 166: English check Figure 3: rename using latin alphabet Figure 4: Increase legend as it is not visible.			gels8040212_makarova	0
+Thanks for your suggestion. In this study, texture profile analysis was investigated by a Texture Analyzer (TA.XT PLUS/50, STABLEMICVO, UK) using a P/0.5R cylindrical probe, and corresponding description has been added to the manuscript in Line 526 marked in red font.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_perova	0
+The gel without the emulsion was taken as the emulsion blank gel, which was the gel formed only by gel continuous phase.	2	1	Line 512: Was 1% found to be within the viscoelastic region?			gels8040212_perova	0
+Wet gel were the emulsion gels we obtained, which had a three-dimensional network structure that can hold a large amount of water. The corresponding Line 570 in the manuscript has been revised to a more accurate expression, emulsion gels.	2	1	Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_perova	0
+It is our great pleasure to get your professional comments. Thanks very much for your contribution to our manuscript. We take the concerns seriously and comments have been carefully addressed.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_perova	0
+Thanks for carefully checking. Whey protein isolate (WPI) has been added in Line 60-61 in the manuscript.	2	1	B1, B2, and B3 looks biphasic, was this the case?			gels8040212_perova	0
+Thanks for your constructive comment. Line 84-88 has been removed and it made the manuscript more logically. Thank you again for the good comment.	2	1	Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels.			gels8040212_perova	0
+The emulsions of different droplet size was prepared at WPI concentration of 1.5 wt% and this information has been added in Line 102. This concentration was selected as a suitable emulsifier concentration based on the results in Figure 1a and 1b, and more details can be found in the manuscript.	2	1	Response 18: Thanks for your carefully check.			gels8040212_perova	0
+Thanks for your good comment. Figure 2 has been modified accordingly to make the legend clearer.	2	1	The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_perova	0
+The manuscript has been revised accordingly in Line 159-161.	2	1	The manuscript is presented in a well-structured manner.The references cited are relevant and current, mostly published within the last three years.The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_perova	0
+Figure 3 has been revised accordingly.	2	1	B1, B2, and B3 looks biphasic, was this the case?			gels8040212_perova	0
+Figure 4 has been adjusted to make the legend clear. In fact, B1, B2 and B3 were in a relatively homogeneous state, and no phase separation had occurred. The shadow in the middle part of the apparent image of the heat-induced gels was because the texture test was carried out before shooting. Unlike TGase-induced emulsion gel had very good elastic texture, the structure of heat-induced gels did not fully recover to its original state due to its soft and less elastic texture after compression.	2	1	Response 1: It is our great pleasure to get your positive comments.			gels8040212_perova	0
+Thanks for your constructive suggestion. Figure 7B has been revised accordingly, that is, a graph was added which displayed the apparent viscosity of all emulsion gels at a fixed shear rate of 0.25/s, showing the differences between groups very visually. Thanks again for your suggestion.	2	1	The article is very well written and only requires few minor changes.			gels8040212_perova	0
+In order to choose appropriate emulsifier, we prepare different WPI concentration of 1.0-2.0 wt% and found the emulsion formulated with 1.5 wt% WPI had smaller size and medimum zeta potential, so it was described to the optimize condition. For clarity, we have also revised the corresponding positions in the manuscript.	2	1	The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_perova	0
+Thanks for your good suggestion. The gel continuous phase was the WPI-gelatin mixture. To more clearly describe the composition of the emulsion gel, Table 3 has been added to the manuscript. The pre-gel mixture, refered to the mixture before gelation occurred, was composed of WPI-gelatin mixture and emulsions, but due to the large number of samples prepared by the single factor experiment, only the general formulation of the emulsion gel was listed in Table 3. And the relevant statements in 4.2.2 and 4.2.3 have been revised in the manuscript.	2	1	The references cited are relevant and current, mostly published within the last three years.			gels8040212_perova	0
+Thanks for your check. It was revised in the manuscript.	2	1	Author Response Point 1: The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			gels8040212_perova	0
+Thanks for your carefully review. The characterization of emulsions was investigated by the droplet size, polydispersity index (PDI) and zeta potential. This expression has been revised in the manuscript.	2	1	The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_perova	0
+PDI was the polydispersity index and the manuscript has been revised in Line 501.	2	1	Line 500: Define PDI at first use Line 507: Clearance height or gap`?			gels8040212_perova	0
+The gap was set to 1 mm. Thank you again for your careful review.	2	1	Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels.			gels8040212_perova	0
+Thanks for your question. 1% strain was within the linear viscoelastic region in our pre-experiment so it was chosen for other tests.	2	1	The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_perova	0
+Thanks for your comment. The statement in 4.2.5 has been revised in the manuscript.	2	1	B1, B2, and B3 looks biphasic, was this the case?			gels8040212_perova	0
+Thanks for your carefully check. Line 533 has been modified which corresponding to the line of revised manuscript.	2	1	One main request would be to make it clearer what was the formulations and conditions tested.			gels8040212_perova	0
+Thanks for your good suggestion and we have improved it.	2	1	Citing the original source is required but so are details needed to reproduce and evaluate the soundness of the manuscript.			genes13030404_makarova	0
+Thanks for your good suggestion and we have added the possible reason in the discussion.	2	1	Blocking IbmiR319a 21 in transgenic sweet potato (MIM319) resulted in a slim and tender phenotype and greater sensitivity 22 to drought stress.			genes13030404_makarova	0
+Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	2011), dis- 44 coloration, abnormal ripening, and so on (Sato, Murakami et al. 2001).			genes13030404_makarova	0
+Thanks for your good suggestions on this point. In our revised manuscript, we have made corresponding modifications to summary our work.	2	1	RNA-Seq results need to be further discussed.			genes13030404_makarova	0
+Thanks for your good suggestion. We removed “and so on” in our revised manuscript.	2	1	As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing.			genes13030404_makarova	0
+Thanks for your reminding. We modified “As we all know” into “More and more research reveals that”.	2	1	The manuscript is well written and the presented evidence supports the conclusions being made.			genes13030404_makarova	0
+It is really true as you mention that the details of methods is needed. But considering that this method is also expressed in thus way in our previous articles (Ren L, et al (2021) Genome-Wide Identification of TCP Transcription Factors Family in Sweet Potato Reveals Significant Roles of miR319-Targeted TCPs in Leaf Anatomical Morphology. Front. Plant Sci. 12: 686698. doi: 10.3389/fpls.2021.686698), we just cite the original source here. Point 4: Pg 6 line 239: I don't understand the point being made here.	2	1	33 Keywords: sweet potato; microRNA319; TCP transcription factor; drought stress.			genes13030404_makarova	0
+Thanks for your question Target mimicry is a common method to study the function of miRNA. There is a non-coding protein gene IPS in Arabidopsis, and its transcription RNA can specifically bind to miRNA through sequence complementarity, but it forms incomplete complementary bubble structure at the cleavage site of miRNA, that is, miRNA cannot cleavage IPS RNA, thus inhibiting the degradation of miRNA to its target gene mRNA. IPS gene is equivalent to the target mimicry. (Franco-Zorrilla et al., 2007) 2 1	2	1	Severe drought restricts crop growth and sig- 38 nificantly reduces yield worldwide (Burke, Lobell et al. 2009), (Hu and Xiong 2014).			genes13030404_makarova	0
+Thanks for your good suggestion and we have improved it.	2	1	Point 4: Pg 6 line 239: I don't understand the point being made here.			genes13030404_perova	0
+Thanks for your good suggestion and we have added the possible reason in the discussion.	2	1	Genes 2022, 13, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Last- name Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations.			genes13030404_perova	0
+Thank you for your positive and constructive suggestions and we have improved it in our revised manuscript.	2	1	As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing.			genes13030404_perova	0
+Thanks for your good suggestions on this point. In our revised manuscript, we have made corresponding modifications to summary our work.	2	1	Point 4: Pg 6 line 239: I don't understand the point being made here.			genes13030404_perova	0
+Thanks for your good suggestion. We removed “and so on” in our revised manuscript.	2	1	RNA-Seq results need to be further discussed.			genes13030404_perova	0
+Thanks for your reminding. We modified “As we all know” into “More and more research reveals that”.	2	1	Response to Reviewer 1 Comments Point 1: The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424.			genes13030404_perova	0
+It is really true as you mention that the details of methods is needed. But considering that this method is also expressed in thus way in our previous articles (Ren L, et al (2021) Genome-Wide Identification of TCP Transcription Factors Family in Sweet Potato Reveals Significant Roles of miR319-Targeted TCPs in Leaf Anatomical Morphology. Front. Plant Sci. 12: 686698. doi: 10.3389/fpls.2021.686698), we just cite the original source here.	2	1	Introduction 36 Drought is a major environmental factor causing abiotic stress (Pearce and RS 2001, 37 Xie, Wang et al. 2017, Meng, Li et al. 2018).			genes13030404_perova	0
+Thanks for your question Target mimicry is a common method to study the function of miRNA. There is a non-coding protein gene IPS in Arabidopsis, and its transcription RNA can specifically bind to miRNA through sequence complementarity, but it forms incomplete complementary bubble structure at the cleavage site of miRNA, that is, miRNA cannot cleavage IPS RNA, thus inhibiting the degradation of miRNA to its target gene mRNA. IPS gene is equivalent to the target mimicry. (Franco-Zorrilla et al., 2007) 2 1	2	1	Blocking IbmiR319a 21 in transgenic sweet potato (MIM319) resulted in a slim and tender phenotype and greater sensitivity 22 to drought stress.			genes13030404_perova	0
+Comments #1 (gene expression assay) and #3 (proper discussion on RNA-Seq data) aren't properly addressed.	3	2	Two members, IbmiR319a and IbmiR319c, were discovered in the miR319 gene family in sweet 19 potato (Ipomoea batatas [L.] Lam).			genes13030404_perova	0
+The colour refer to the SEEBASE depth to basement, the legend has been added • Figure 2: The left spine (or border) does not appear on the pdf reviewed.	2	1	• Lines 339-357/Section 4.3: The map shown on Figure 9 that provides the location where groundwater and springs were sampled could be referred to.			geosciences12010037_makarova	0
+Thanks, the figure have been entirely revisited and divided into four subfigures.	2	1	Helium content of the aquifer systems was sampled and measured.			geosciences12010037_makarova	0
+The figure has been redrawn and the magnetic lineaments indicated • Figure 4: This figure could go in the appendix.	2	1	It is good to let the reader know that not all seismic surveys provided the same quality, but this is more a technical point.			geosciences12010037_makarova	0
+Thank you. We hesitated on this point but decided to keep the figure in as the quality of the seismic in the shallow level is really crucial to the discussion. We need to know if the interpretation is reliable, especially in this shallow zone that is not well imaged by this type of data. A misinterpretation can led to the targeting of circulation pathways that are in fact only artefacts.	2	1	Interpretation of shallow horizons and faults using all publicly available geological and geophysical data of the Beetaloo region.			geosciences12010037_makarova	0
+We removed the seismic interpretation of figure 7G (cross) that were misleading the reader and add the interpretation of the fluid leakage features on the images	2	1	Results appear to be sound and supported by the illustrations.			geosciences12010037_makarova	0
+this is due to the different seismic acquisition technics and processing applied in each survey as well as the fact that our interval of interest is shallow and not fitting with the interval of interest the companies tried to best image with those datasets”	2	1	done Figure 4 -Poor quality figure (low resolution) in the version available to the reviewer.			geosciences12010037_makarova	0
+We added “as the presence of fluid or gas can locally alter the seismic signal”	2	1	This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control.			geosciences12010037_makarova	0
+We added: “We define as recent tectonic activity the events that are recorded at the surface and can alter the surface with active tectonic features, such as modifications of the drainage system” • Discussion: One general comment, could the use of other tracers (radon, of stable isotopes of water) yield also insight on the origin of the fluids sampled in this study?	2	1	Overall, the implications of this study are important not only for industrial purposes (reservoir exploration and operation) but also for our basic understanding of subsurface flow processes over geological timescales.			geosciences12010037_makarova	0
+Radon cannot give this information since with the short half-live of 3.8days it's signal comes only from the direct vicinity of the sampled location. Stable isotopes of water could in principle do this if there is a geothermal signature on them (such as plotting far left to the meteoric water line - see inset in Figure 1 of McIntosh and Ferguson, 2020 – Geophysical research Letter) - but we have not seen such a signature. We may mention that in the text - but for such a signature to be discernible the bulk of the water needs to be geothermal, and what we see is only a small admixture.	2	1	These often highlight tectonic accommodation space creation better than maps.			geosciences12010037_makarova	0
+The revisited figure 43 is now providing this information Seismic reflection 2D lines (1989-2015).	2	1	The replies to detailed comments are in purples, directly below each comment Frery et al.			geosciences12010037_makarova	0
+Thank you, the details comments on those figures have been followed and new figures are provided Helium measurements from groundwater samples.	2	1	Consider replacing well names on the map with numbers at the well position (white circles, black typeface) and add a table in the legend listing number and corresponding name of the well.			geosciences12010037_makarova	0
+"There is no other reference to date to the ""primary documentation"" but the link to the data (they should now be online) which are in the fact sheets."	2	1	These often highlight tectonic accommodation space creation better than maps.			geosciences12010037_makarova	0
+We have redone the figure 2 and added explanations on the process (see reply to comments on figure 2). Figure 8 should now be easier to read The surface drainage network line dataset was indeed based on the simplified (n=130) network following the following approach: Small segments with less than 10 nodes were automatically removed in SKUA (Paradigm/Emerson Trademark) and one iteration of smoothing was applied that further removed small irregularities without changing the overall orientation of major drainage lines.	2	1	Results appear to be sound and supported by the illustrations.			geosciences12010037_makarova	0
+There is indeed an error on the figure referencing, we modified to Figure 3B where the lineaments are now clearly indicated.	2	1	Reference is made to “…prominent […] magnetic lineaments which can be caused by faulting”, and “…several circular low magnetization anomalies, approximately two kilometers in diameter, aligned with northwest trending lineaments”.			geosciences12010037_makarova	0
+We indeed need to add this link- we will put a link in place upon acceptance of the paper Figures Figure 1A.	2	1	Review summary Scientific aims clearly stated.			geosciences12010037_makarova	0
+done, we also modified the fault legend to add clarity - Replace white labels with black lettering with no labels and white lettering.	2	1	Quality improved - Very, very overloaded figure, screen-dump from a mapping programme?			geosciences12010037_makarova	0
+We separated the legends from figures 1A and 1B as to add clarity. This article is focused on the Beetaloo subbasin and we use the direct input for Geoscience Australia in this stratigraphic column. We are not willing to add complexity by presenting in detail the Basins that are covering the Beetaloo Sub-Basin. We removed from the legend the details on the different basins as we understand that it is leading the reviewer in directions we are not exploring on the CLA stratigraphy- we do not refer this complexity in the results and discussion.	2	1	- Add scale. done - Add colour scale for map (depth).done -Well- and fault-labels largely illegible.			geosciences12010037_makarova	0
+Quality improved - Very, very overloaded figure, screen-dump from a mapping programme?	2	1	The accuracy of the resulting plot can therefore be questioned.			geosciences12010037_makarova	0
+Thanks, the figure have been entirely revisited and divided into four subfigures as advised below -Consider splitting into several maps, e.g.	2	1	If the latter applies, make sure to provide a scale.			geosciences12010037_makarova	0
+This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control. In this process, minor mismatches of the automated simplified lines were removed and small drainage segments were connected along major lineaments for further simplification and reduction of the dataset. The adopted approach was: Small segments with less than 10 nodes were automatically removed in SKUA (Paradigm/Emerson Trademark) and one iteration of smoothing was applied that further removed small irregularities without changing the overall orientation of major drainage lines. We added this description of the adopted approach in the legend Figure 3 -Very small/illegible typeface on map coordinates – consider simplifying scales and use larger typeface.	2	1	the seismic interpretation of figure 7G (cross) that were misleading the reader and add the interpretation of the fluid leakage features on the images • Figure 9: Remove ‘s’ from <10 yearss.			geosciences12010037_makarova	0
+figure redrawn - Replace white labels with black typeface with white typeface.	2	1	This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control.			geosciences12010037_makarova	0
+Well label typos modified -Thin grey seismic lines are not explained in the legend or caption.	2	1	The study concludes that there the shallow aquifers are connected to deep-seated gas source along faults, some of which show evidence of recent movement.			geosciences12010037_makarova	0
+Legend added for those lines -The outline of the Eastern and Western Beetaloo Basin in Figure 3 and 4 should be kept identical in order to position Figure 6 A, B and C in relation to the seismic lines shown in Figure 4.	2	1	The map differentiates between “BaseCambrian seismic faults”, Post-Wilton “Strike-slip” and “Reverse” faults, and “FAULTS”.			geosciences12010037_makarova	0
+sochores added as supplementary material	2	1	Also, increase the thickness of the fault lines to make them more obvious.			geosciences12010037_makarova	0
+done, legend cleared from information not represented	2	1	Although this is described in the caption, it is not easy to grasp for someone not familiar with the stratigraphy in the area without spending time some time with paper and pencil.			geosciences12010037_makarova	0
+checked and clarified Clean up map: 1.	2	1	Thank you, we truly appreciated the time took by the reviewer to provide detail guidance to improve the manuscript and the figures			geosciences12010037_makarova	0
+Clearly identified in the legend and representation 2.	2	1	Quality improved - Very, very overloaded figure, screen-dump from a mapping programme?			geosciences12010037_makarova	0
+removed	2	1	Reference is made to “…prominent […] magnetic lineaments which can be caused by faulting”, and “…several circular low magnetization anomalies, approximately two kilometers in diameter, aligned with northwest trending lineaments”.			geosciences12010037_makarova	0
+Outline size modified and homogenised with other figures 4.	2	1	Depth conversion using check-shot velocities from 26 wells.			geosciences12010037_makarova	0
+Red selected 5.	2	1	Section 4.3 Helium concentrations and isotopic composition measured in groundwater, is framed around results from previous studies.			geosciences12010037_makarova	0
+modified	2	1	Consider replacing well names on the map with numbers at the well position (white circles, black typeface) and add a table in the legend listing number and corresponding name of the well.			geosciences12010037_makarova	0
+modified accordingly Consider adding a simple conceptual sketch highlighting the stratigraphic position of the different faults (and possible fluid escape structures– this would also help visualizing which stratigraphic intervals the potential fluid escapes originate from and which stratigraphic intervals are potentially connected along potential fault-related fluid flow pathways.	2	1	).done, legend cleared from information not represented 4. d) Explain/improve labels (what do you mean by “ntspr_2M_gw”?			geosciences12010037_makarova	0
+With the new classification the reader should be able to better read the paper and can now directly refer to the stratigraphic table Figure 8.	2	1	One of the key issues in the Beetaloo Sub-basin, the risk of groundwater contamination from fracking-based production of the unconventional plays.			geosciences12010037_makarova	0
+Done, figure clarified as well Figure 9.	2	1	A misinterpretation can led to the targeting of circulation pathways that are in fact only artefacts.			geosciences12010037_makarova	0
+done, we also totally revisited this figure as to have a cleared output	2	1	If the primary documentation is not accessible, the authors should state this clearly.			geosciences12010037_makarova	0
+The helium measurements have been added as supplementary data and the author made a clear difference between this study Helium measurements and other tracer studies we are citing.	4	1	Figures, however, need to be significantly improved for the final manuscript (see detailed comments below).			geosciences12010037_makarova	0
+Done, thank you	4	1	It would certainly help if they were highlighted directly on the figure.			geosciences12010037_makarova	0
+The isochores have been added in the seismic supplementary dataset	4	1	Conclusions appear supported by the observations made in this study, but presentation of results needs to be improved.			geosciences12010037_makarova	0
+Post Wilton “Strike-slip” and “Reverse” faults have been redrawn in all figures (Figure 1, 7, 9) with a new colour code, the fluid escape features are now shown as red dots in the legend as well.	4	1	Author Response Thank you for your detailed review.			geosciences12010037_makarova	0
+Number of measurements added in each plot. Figure 2a correspond to the SEEBASE structural lineaments (n=681).	4	1	Review summary Scientific aims clearly stated.			geosciences12010037_makarova	0
+we are very sorry for the mistake, and the manuscript was modified accordingly.	2	1	"The authors report experience from their surgical center with performing abdominoplasty in women with ""thrombophilia"" which is not clearly defined."			healthcare10030476_makarova	0
+The criticism of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	The results are of interest to both surgeons and haematologists/haemostaseologists.			healthcare10030476_makarova	0
+The observation of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	Severity of thrombophilic states are now yet well categorized, therefore more clarification needs to be provided here.			healthcare10030476_makarova	0
+The observation of the reviewer has been accepted and the table 1 was modified accordingly.	4	1	Author Response Reviewer(s)' Comments to Author:Reviewer #1: Verolino et al.			healthcare10030476_makarova	0
+The new manuscript has been evaluated by an expert of English language.	4	1	2-3% of what for the 20210 polymorphism and 3-8% of what for FV Leiden?			healthcare10030476_makarova	0
+The observation of the reviewer has been accepted and the table 1 was modified accordingly.	4	1	The results are of interest to both surgeons and haematologists/haemostaseologists.			healthcare10030476_perova	0
+The new manuscript has been evaluated by an expert of English language.	4	1	in Sode et al CMAJ 2013 doi: 10.1503/cmaj.121636.			healthcare10030476_perova	0
+we are very sorry for the mistake, and the manuscript was modified accordingly.	2	1	However, I have several concerns which must be adequately addressed before the manuscript is suitable for publication.			healthcare10030476_perova	0
+The criticism of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	Also, “20210 gene” should be “20210 polymorphisms, or “G20210A polymorphism” to be completely correct.			healthcare10030476_perova	0
+The observation of the reviewer has been accepted and the manuscript was modified accordingly.	2	1	"In line 89 authors state that ""thrombophilic states are rather venous than arterial""."			healthcare10030476_perova	0
+The quoted literature has been adapted to the ACS Style Guide. For that, the authors used to software package Zotero.	2	1	I suggest to re-write the aims and study questions more clearly.			healthcare10040723_perova	0
+STD are included in the all tables were indicated: sum+STD has been added in the legend.	2	1	1 Review comments Title: Medical students' perceptions towards digitization and artificial intelligence: A mixed methods study Thank you for the opportunity to review this manuscript.			healthcare10040723_perova	0
+In Table 3 (=) has been changed into (0). It was a typo, sorry.	2	1	I suggest the authors to include a short section including the implications of the study after the conclusions.			healthcare10040723_perova	0
+Line 252: 7/24 has been changed into 24/7.	2	1	Authors have sufficiently described the study methodologies including the survey and drawn The following revisions should be addressed before publication:			healthcare10040723_perova	0
+Line 267 and 268, the description of figure 1 “focused on senses” means that the diagnostic workup should be more focused on the physical examination (the doctor’s senses) rather than based on technical diagnostics. In the revised manuscript has been added in line 270 “and use their senses in physical examinations rather than rely on impersonal technical tools for the diagnostic work-up”.	2	1	Dear Authors, Here attached, please find my comments for the improvement of your paper.			healthcare10040723_perova	0
+In Line 266/267 the term “undermine” was wrong. It has been changed into “ease”. Thank you for the hint.	2	1	Also, the research questions lack describing the study setting.			healthcare10040723_perova	0
+Table 5: The students could rank their answer. 1 = don’t know, 2= false up to 7 = fully agree. The statistical calculation was done only for ranks 2 to 7, excluding the students, who could not or didn’t want to answer the question. The authors knew from the qualitative study part, that some students had no knowledge about AI. Therefor the “don’t know” option was included. In the first draft of the manuscript the legends contained a mistake, because the “fully agree” option had the highest number (7) but not 3. This has been changed in the revised manuscript.	2	1	It needs the careful wording of the study participants.			healthcare10040723_perova	0
+2. The reviewer discussed in much detail which design might better describe the method used in our study. In the sequential exploratory strategy a qualitative component is followed by a quantitative component (Creswell, JW. ; Plano Clark, VL. Designing and Conducting Mixed Methods Research. Thousand Oaks, CA: Sage; 2011). Aspects of the organizational environment (e.g., creative problem solving) were identified in the qualitative component and statistically confirmed or rejected in the preceding quantitative study phase, which is exactly what the authors did in this study. However, the reviewer suggested that the convergent parallel design might fit our methodology even better. In the convergent parallel strategy, quantitative and qualitative data collection occurred concurrently. The components are given equal weight, and the two datasets are analyzed and compared in parallel. Quantitative data identified factors statistically, which were integrated with qualitative data from in depth interviews with a subset of participants to understand how and why the identified factors influenced participants’ attitudes. In the method’s section of our study we lined out that the results of the first qualitative study part were the basis for the development of the online survey. Based on the qualitative study part, the questions for the qualitative study were formulated when students’ attitudes were expressed frequently. In this sense, the first (quantitative) phase informs the next, but the reviewer is right, that both phases are interlinked. Although our study integrates components of both strategies, the authors still think, that the sequential exploratory strategy describes our methods better than the convergent parallel design (see also: Curry et al Circ. Cardiovasc, Qual. Outcomes 2013, 6:119-123). As suggested by the reviewer, the authors specified in the method section that we used an additive rather than a parallel integrative strategy.	2	1	I think, the manuscript benefits from a minor language editing.			healthcare10040723_perova	0
+The manuscript has been changed accordingly.	2	1	Also, the research questions lack describing the study setting.			healthcare10040723_perova	0
+The authors followed the reviewer’s suggestions and re-wrote this para as follows: “The overall objective of this study was to investigate the today’s medical students attitudes towards AI and other digital working tools. We wanted to understand if age, gender, semester level and curriculum type influences their views. This study also assembled information on students’ understanding of AI algorithms and digital applications in health care and assessed their level of confidence in working alongside these tools after graduation in patient care. It is our belief that this information may possess the means to employ digital tools including AI into the curriculum of medical students efficiently, enhancing their confidence in using them and therefor to better equip our future physicians with sufficient knowledge.” On page 2, line 111, the phrase ‘to affirm or dismiss,’ does not make sense.	2	1	I suggest to remove the phrase ‘so called,’ that has been used in the conclusion section or any other section.			healthcare10040723_perova	0
+The phrase in question has been replaced by “to confirm”.	2	1	However, there are some issues to be addressed before recommending this paper for publication.			healthcare10040723_perova	0
+The authors choose 4 pre-clinical and 4 clinical students. The intention was - as outlined in the initial manuscript version - “… to understand how they perceive the subject of interest”. The wording has been specified, to clarify that in total 8 students tested the items but not 4 as the reviewer assumed. This number was considered sufficient a) to detect inconsistencies and b) to verify the correct meaning of the questions. AG and JE are two of the authors (first letter of the first and last name). In order to avoid any misunderstanding, this has been changed to “the authors” in the revised manuscript.	2	1	I suggest the authors to replace this with appropriate phrase.			healthcare10040723_perova	0
+Convenience sampling is defined as a method to collect data from a conveniently available pool of respondents. We used it, because it is the most commonly used sampling technique as it's incredibly prompt, uncomplicated, and economical.	2	1	I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.			healthcare10040723_perova	0
+The captation has been changed to “Characteristics of the study cohort.	2	1	I suggest the authors to include a short section including the implications of the study after the conclusions.			healthcare10040723_perova	0
+The sampling procedure was clearly described in the initial manuscript. For phase 1: “All participants of the first phase were students from from 1st to 6th year (undergraduate 1st to 2th year, graduate 3th to 6th year) from German Universities. The inclusion criteria were to actively study medicine and their agreement for their voluntarily participation. In the same way, the exclusion criteria were to have suspended their studies, as well as other exceptional situations. Prior to start, informed consent was obtained, which was followed by the collection of telephone numbers and email addresses. They were selected purposely and consecutively, in part by snowball until theoretical saturation was reached.” For phase 2: “For the second, quantitative study phase identical inclusion/exclusion criteria applied.” This referred to medical students of all semesters, excluding students from other faculties. “The online survey was sent to all medical faculties in Germany from which most forwarded the survey invitation by email to about 80 000 medical students to fulfill the principle of maximum diversity through convenience sampling method. Each contained an invitation letter and an information sheet.” Surely, sampling procedures in mixed method studies are phase depended different.	2	1	I suggest the authors to replace this with appropriate phrase.			healthcare10040723_perova	0
+The statement has been changed into: “The focus on German educational system and the fact that only a small fraction of the total number of medical students filled out the online survey makes a generalization of the answers difficult”.	2	1	‘The items were then tested through a pilot study consisting of a 117 group of 4 pre- and clinical students, mediated by AG and JE’.			healthcare10040723_perova	0
+The authors agree and the phase “so called” has been removed.	2	1	In the abstract, lines 23-27, I suggest to use the following expression or something like that.			healthcare10040723_perova	0
+The authors agree and this statement has been re-written in the revised manuscript version.	2	1	Page 12, line 389, the statement which says, ‘The non-probabilistic sampling makes a generalization of the answers difficult,’ is confusing.			healthcare10040723_perova	0
+The authors fully agree. Therefore, the conclusion paragraph has been split into the “Conclusion” paragraph, in which the study’s conclusions has been outlined, and into a the new para “Implications” where the usefulness of the integration of eHealth aspects (including AI) into the curriculum is discussed.	2	1	In the methods section, clearly describe the sampling procedure for the quantitative and qualitative study parts separately.			healthcare10040723_perova	0
+We have modified this part in revised manuscript (Line 233-236). This sentence has been changed to “A suspension of A. rhizogenes strain GV3101 containing the pORER4-OfSPL11-GFP recombinant plasmid was aspirated using a 1 mL range syringe (needle not included), and the suspension was gently injected into the abaxial surface of four-week-old tobacco leaves avoiding the leaf veins.” Point 2: Rephrase lines 266-268 is unclear; wording, grammar and punctuation are wrong.	2	1	"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 38).	2	1	"Point 27: In line 395 replace ""it revealed"" with revealed."			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we have replaced this word in the revised version (Line 63).	2	1	"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 65)	2	1	A deeeping on protein an nucleotide structures was also reported.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 69).	2	1	"Point 26: In line 318 ""We"" must be lowercase."			horticulturae8050412_makarova	0
+We had made correction in the manuscript (Line 83-84). The statements of “RNA extractions” were corrected as “total RNA was extracted using the RNA prep Pure Plant Plus Kit (Tiangen, Beijing, China).” Point 10: In line 91 what brand was Taq polymerase from?	2	1	Point 4: Line 267-268: Please, check this sentence.			horticulturae8050412_makarova	0
+We had made correction in the manuscript (Line 92). Taq polymerase was purchased from TaKaRa Biotechnology, Inc, Dalian, China.	2	1	Point 10: Figure 9 caption: measure instead of measuring.			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 102-103). The pORER4-GFP vector and the plant expression vector OfSPL11-GFP were purchased from Tsingke, Biotechnology, Inc, Hangzhou, China.	2	1	Point 4: Line 267-268: Please, check this sentence.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction carefully (Line 115 and 270).	2	1	"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 119 and 276).	2	1	"Point 7: In line 65 replace ""And"" with In addition."			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 129).	2	1	"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 136).	2	1	Therefore, we suggest that salt stress can induce the activity of the OfSPL11 promoter.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction carefully (Line 138).	2	1	Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 145).	2	1	Results clearly describe the protein localization as well as its role in stress response.			horticulturae8050412_makarova	0
+"We had made correction in the manuscript (Line 156-157). The statements of “the MDA assay kit” were corrected as “the MDA assay kit (Jiancheng Bioengineering Institute, Nanjing, China).” Point 20: In line 154 add ""strain"" after receptor and mention kit brand Response 20: Thank you for your comment."	2	1	"Point 27: In line 395 replace ""it revealed"" with revealed."			horticulturae8050412_makarova	0
+"We had made correction in the manuscript (Line 162 and 163). The statements of “the yeast Y187 receptor strain preparation and transformation kit” were corrected as “the yeast Y187 receptor strain preparation and transformation kit (Coolaber, Beijing, China).” Point 21: In line 169 add ""were done"" at the end."	2	1	A deeeping on protein an nucleotide structures was also reported.			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 177).	2	1	In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions.			horticulturae8050412_makarova	0
+Thank you for your comment, we had made correction in the manuscript (Line 270).	2	1	"Point 27: In line 395 replace ""it revealed"" with revealed."			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 271).	2	1	"Point 16: In line 129 replace ""strains and ""strain"" with lines and line."			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction carefully (Line 273).	2	1	We have modified this part in revised manuscript (Line 121-125).			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 301).	2	1	This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction carefully (Line 327).	2	1	"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 403).	2	1	Response 10: Special thanks to you for your good suggestion.			horticulturae8050412_makarova	0
+Thank you for your comment, we had made correction in the manuscript (Line 415).	2	1	"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+We have modified this part in revised manuscript (Line 326-328).	2	1	The LUC signal increased profoundly after subjection to HS (37℃), although under normal conditions (room temperature), the LUC signal was very low, which implied that HS activated the promoter activity of LlWRKY39.			horticulturae8050412_makarova	0
+genes. We feel sorry for the unclear statement. In our study, we used the amino acid sequence of OfSPL11 to compute the protein model. Secondary structure predictions were conducted on the PRABI (http://www.prabi.fr/spip.php?page=sommaire) protein structure prediction server. The ExPASy (http://web.expasy.org) was used to analyze the molecular weight, theoretical isoelectric point, and other physiochemical properties of the translated protein. Similar methods of operation and efforts can be found in other articles (Islam, 2021). Once again,we sincerely thank you for the recommended method, which we will use in future studies. Islam, M.D.; Rahman, M.M.; Rahman, M.M. ; Jin, X.J. ; Sun, L.L. ; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S. ; Zhang, W.J. ; Sun, D.Z. In Silico and Transcription Analysis of Trehalose-6-phosphate Phosphatase Gene Family of Wheat: Trehalose Synthesis Genes Contribute to Salinity, Drought Stress and Leaf Senescence. 2021, 12, 1652. https://doi.org/10.3390/genes12111652 Point 2: Auhtors report that Actin was used as housekeeping gene.	2	1	In this view, I've only some few questions for the authors.			horticulturae8050412_makarova	0
+We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015). Dheda, K.; Huggett, J.F. ; Bustin, S.A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR. Biotechniques. 2004, 37: 112-119. https://doi.org/10.2144/04371RR03 Czechowski, T.; Stitt, M.; Altmann, T. Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol. 2005, 139: 5-17. https://doi.org/10.1104/pp.105.063743 Hou, H.; Jia, H.; Yan, Q.; Wang, X. Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance. Int. J. Mol. Sci. 2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X. ; Wang, Y.G. ; Bao, Z.Y. ; Zhao, H.B. Identification of Suitable Reference Genes for Gene Expression Normalization in the Quantitative Real-Time PCR Analysis of Sweet Osmanthus (Osmanthus fragrans Lour.). Plos one. 2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.	2	1	Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 130).	2	1	Point 4: Line 267-268: Please, check this sentence.			horticulturae8050412_makarova	0
+We have revised the manuscript according to your comments (Line 274-277). This sentence has been changed to “A. tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 3: Rephrase lines 317-318.	2	1	"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+We had made correction carefully (Line 283).	2	1	"Point 25: In line 292 ""The"" must be lowercase."			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 310).	2	1	The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 319).	2	1	"Point 27: In line 395 replace ""it revealed"" with revealed."			horticulturae8050412_makarova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 324).	2	1	Response 4: We feel sorry for the unclear statement.			horticulturae8050412_makarova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 366).	2	1	In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion. We had made correction carefully (Line 381).	2	1	"Point 12: Line 446: ""..participate in salt stress"" Response 12: Thank for your kindly suggestion."			horticulturae8050412_makarova	0
+Special thanks to you for your good suggestion, we had made correction in the manuscript (Line 398).	2	1	Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			horticulturae8050412_makarova	0
+"Thank for your kindly suggestion. We have revised the manuscript according to your comments (Line 456). The statements of “participate in salt stress responses” were corrected as “participate in salt stress.” Point 12: Line 448: check the word ""redycing"" Response 12: Thank you for your comment."	2	1	Response 4: We feel sorry for the unclear statement.			horticulturae8050412_makarova	0
+We have modified this part in revised manuscript (Line 233-236). This sentence has been changed to “A suspension of A. rhizogenes strain GV3101 containing the pORER4-OfSPL11-GFP recombinant plasmid was aspirated using a 1 mL range syringe (needle not included), and the suspension was gently injected into the abaxial surface of four-week-old tobacco leaves avoiding the leaf veins.” Point 2: Rephrase lines 266-268 is unclear; wording, grammar and punctuation are wrong.	2	1	"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_perova	0
+We have modified this part in revised manuscript (Line 326-328). This sentence has been changed to “To better understand why OfSPL11 transgenic lines exhibit enhanced abiotic stress tolerance, we analyzed the expression of known abiotic stress response genes in 3-week-old wild-type and transgenic Arabidopsis.” Point 4: In lines 271-272 and 395-396, how do you explain that salt induced promoter trans-activation?	2	1	; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S.			horticulturae8050412_perova	0
+In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China). The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves. The LUC signal increased profoundly after subjection to HS (37℃), although under normal conditions (room temperature), the LUC signal was very low, which implied that HS activated the promoter activity of LlWRKY39. In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions. Therefore, we suggest that salt stress can induce the activity of the OfSPL11 promoter.	2	1	"Point 12: Line 446: ""..participate in salt stress"" Response 12: Thank for your kindly suggestion."			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 38).	2	1	"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			horticulturae8050412_perova	0
+Thank for your kindly suggestion, we have replaced this word in the revised version (Line 63).	2	1	"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 65).	2	1	"Point 24: In line 264 replace ""indicate"" with indicated."			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 69).	2	1	"Point 24: In line 264 replace ""indicate"" with indicated."			horticulturae8050412_perova	0
+We had made correction in the manuscript (Line 83-84). The statements of “RNA extractions” were corrected as “total RNA was extracted using the RNA prep Pure Plant Plus Kit (Tiangen, Beijing, China).” Point 10: In line 91 what brand was Taq polymerase from?	2	1	Point 6: Line 302: plant leaves instead of plants leaves.			horticulturae8050412_perova	0
+We had made correction in the manuscript (Line 92). Taq polymerase was purchased from TaKaRa Biotechnology, Inc, Dalian, China.	2	1	"Point 23: In line 262 replace ""were treatment by"" with were treated with."			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 102-103). The pORER4-GFP vector and the plant expression vector OfSPL11-GFP were purchased from Tsingke, Biotechnology, Inc, Hangzhou, China.	2	1	Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction carefully (Line 115 and 270).	2	1	"Point 24: In line 264 replace ""indicate"" with indicated."			horticulturae8050412_perova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 119 and 276).	2	1	"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			horticulturae8050412_perova	0
+We have modified this part in revised manuscript (Line 121-125). This sentence has been changed to “Symbiotic medium was prepared by weighing 4.414 g MS, 20 g sucrose, 10 g glucose, 7 g agar into a conical flask, sterile water was fixed to 1 L. The medium was sterilized in an autoclave for 30 min and the temperature was lowered to 55℃, add 100 μl naphthalene acetic acid (0.1 mg L-1), 500 μl zeatin (1mg L-1), 1 mL folic acid (1mg L-1), 1 mL 6-BA (2mg L-1), 500 μl acetosyringone (0.1 mol L-1).” Point 15: In line 123 delete the first phrase which is repetitive (was already written before in Line 100).	2	1	"Point 25: In line 292 ""The"" must be lowercase."			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 129).	2	1	Point 18: In line 138 Seedling should be lowercase.			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion, we had made correction carefully (Line 136).	2	1	A deeeping on protein an nucleotide structures was also reported.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction carefully (Line 138).	2	1	2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 145).	2	1	Point 10: Figure 9 caption: measure instead of measuring.			horticulturae8050412_perova	0
+"We had made correction in the manuscript (Line 156-157). The statements of “the MDA assay kit” were corrected as “the MDA assay kit (Jiancheng Bioengineering Institute, Nanjing, China).” Point 20: In line 154 add ""strain"" after receptor and mention kit brand Response 20: Thank you for your comment."	2	1	Response 9: Thank for your kindly suggestion.			horticulturae8050412_perova	0
+"We had made correction in the manuscript (Line 162 and 163). The statements of “the yeast Y187 receptor strain preparation and transformation kit” were corrected as “the yeast Y187 receptor strain preparation and transformation kit (Coolaber, Beijing, China).” Point 21: In line 169 add ""were done"" at the end."	2	1	In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			horticulturae8050412_perova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 177).	2	1	Point 6: Line 302: plant leaves instead of plants leaves.			horticulturae8050412_perova	0
+Thank you for your comment, we had made correction in the manuscript (Line 270).	2	1	"Point 24: In line 264 replace ""indicate"" with indicated."			horticulturae8050412_perova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 271).	2	1	In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction carefully (Line 273).	2	1	In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 301).	2	1	"Point 8: Line 315: maybe a semicolon after ""salt stress"" is appropriate."			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction carefully (Line 327).	2	1	"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_perova	0
+Thank for your kindly suggestion, we had made correction carefully (Line 403).	2	1	Do the authors have performed an efficiency test before using it?			horticulturae8050412_perova	0
+Thank you for your comment, we had made correction in the manuscript (Line 415).	2	1	"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_perova	0
+We feel sorry for the unclear statement. In our study, we used the amino acid sequence of OfSPL11 to compute the protein model. Secondary structure predictions were conducted on the PRABI (http://www.prabi.fr/spip.php?page=sommaire) protein structure prediction server. The ExPASy (http://web.expasy.org) was used to analyze the molecular weight, theoretical isoelectric point, and other physiochemical properties of the translated protein. Similar methods of operation and efforts can be found in other articles (Islam, 2021). Once again,we sincerely thank you for the recommended method, which we will use in future studies.	2	1	In this view, I've only some few questions for the authors.			horticulturae8050412_perova	0
+We tested the amplification efficiency before using housekeeping genes. Referring to the previous study of Arabidopsis housekeeping genes (Dheda, 2004; Czechowski, 2005; Hou, 2018), we selected Atactin1 (F: AGGCACCTCTTAACCCTAAAGC, R: GGACAACGGAATCTCTCAGC), Atactin2 (F: TTGTGCTGGATTCTGGTGATGG, R: CCGCTCTCTGCTGTTGTGGTG) and Atactin8 (F: GAATTACCCGACGGACA, R: ACGGTCTGCAATACCT) were tested for efficiency, and we found that the best result was Atactin2. The best housekeeping gene for O. fragrans is OfACT, which has been reported in a previous study (Zhang, 2015).	2	1	In Silico and Transcription Analysis of Trehalose-6-phosphate Phosphatase Gene Family of Wheat: Trehalose Synthesis Genes Contribute to Salinity, Drought Stress and Leaf Senescence.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 130).	2	1	2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X.			horticulturae8050412_perova	0
+We have revised the manuscript according to your comments (Line 274-277). This sentence has been changed to “A. tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 3: Rephrase lines 317-318.	2	1	Point 18: In line 138 Seedling should be lowercase.			horticulturae8050412_perova	0
+We are very sorry for our negligence, we had made correction in the manuscript (Line 366).	2	1	"Point 23: In line 262 replace ""were treatment by"" with were treated with."			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion. We had made correction carefully (Line 381).	2	1	2018, 19, 940. https://doi.org/10.3390/ijms19040940 Zhang, C.; Fu, J.X.			horticulturae8050412_perova	0
+Special thanks to you for your good suggestion, we had made correction in the manuscript (Line 398).	2	1	"Point 26: In line 318 ""We"" must be lowercase."			horticulturae8050412_perova	0
+"Thank for your kindly suggestion. We have revised the manuscript according to your comments (Line 456). The statements of “participate in salt stress responses” were corrected as “participate in salt stress.” Point 12: Line 448: check the word ""redycing"" Response 12: Thank you for your comment."	2	1	This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			horticulturae8050412_perova	0
+Thank you for your comment. We are very sorry for our negligence. We had made correction carefully (Line 457). The word of “redycing” were corrected as “reducing.” 2 1	2	1	"It ends with ""then"" Response 4: Thank you for your comment."			horticulturae8050412_perova	0
+Thanks for your reminder. In our study, 66 children were taken part in our analysis. But we failed to get all children’s feces and urine sample every day. Thus, we collected sample number was less the involved 66 children.	2	1	The digestion process is repeated until the sample becomes sticky.			ijerph19127332_makarova	0
+Thank you for your reminder. We have uniformed the units to “(mL/d)” Point 4.	2	1	We have added the description of sampling site in the materials and methods.			ijerph19127332_makarova	0
+At present, the related content in manuscript has been revised to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Heavy metal elements accumulate in the 36 human body and interfere with the human endocrine system [5], damage the body's car- 37 diovascular and nervous systems [6.7], and can even lead to cancer [8].			ijerph19127332_makarova	0
+In the manuscript, the data is with outlier. However, we used the data without outlier to construct the figure. The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 5. At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	The site we studied is an e‒waste dismantling area with severe soil pollution that is typical of e‒waste dismantling areas in South China.			ijerph19127332_makarova	0
+We have reorganized the SM figures and the manuscript content. The modifications are as follows: “The frequency distribution histograms show few outliers, most being high values (Figure S3 (a) and Figure S4). Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers. Other outliers may be due to the behavior of certain children, such as pica behavior or spending unusually large amounts of time playing in grassland. The medians of the SIR values after removing the high values (see Figure S3(b) and Figure 1) were taken as the final SIR values for children living in the studied e‒waste dismantling site in South China.” Point 7.	2	1	In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn.			ijerph19127332_makarova	0
+It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5). However, since negative SIR values are physically meaningless, the RI of the SIR for children living in e‒waste dismantling sites is 0‒730.4 mg/d. In this study, 95th per-centile values (383.3 mg/g) would be as recommended value.” Point 8.	2	1	Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Point 2.			ijerph19127332_makarova	0
+It has been changed to “The 95th HQ values based on SIR mean value decreased in the order of Pb > As > Cr > Cu > Ni > Zn > Cd and all of them were below 1.” Point 9.	2	1	Title- ‘in’ should be used for larger areas, can you add the study locations?			ijerph19127332_makarova	0
+We have revised all the figure captions in SM.	2	1	Lines 209-211: Different SIR values between manuscript and figure 1.			ijerph19127332_makarova	0
+Thanks for your advice, we have made the changes to the title following:” Estimation of Children’s Soil/Dust Ingestion Rates and Health Risk at E‑Waste Dismantling Area”	2	1	Lines 176-178: Different units between manuscript and SM (figure S1).			ijerph19127332_makarova	0
+Thanks for your suggestion. We have introduced the sampling sites and added the following information: “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428 Point 3.	2	1	The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C.			ijerph19127332_makarova	0
+Thanks for your suggestion, we have sorted out the method part again, and now the modification is as follows: “Feces samples and food samples were pretreated in the same way. The digestion process was repeated until the sample becomes sticky.	2	1	Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.			ijerph19127332_makarova	0
+For this question, our answer is following: this data is from the Technical Guidelines for Soil Pollution Risk Assessment of Construction Land in China. The website address of the guidelines is https://www.mee.gov.cn/ywgz/fgbz/bz/bzwb/trhj/201912/W020191224560850148092.pdf	2	1	Point 1、This study estimated the heavy metal exposure to children through soil and dust Ingestion in electronic waste (e-waste) dismantling areas.			ijerph19127332_makarova	0
+For this problem, we conducted a questionnaire survey when collecting samples before the experiment, and all the guardians of the sample providers knew and agreed to the experiment.	2	1	The conditions of microwave digestion are the same as mentioned above.			ijerph19127332_makarova	0
+Collecting samples followed the USEPA recommendation of a period of 28 hours from food to feces and urine. Thus, we cannot provide the data for each day. And we also revised the table 1 legend (removing the “each day”).	2	1	S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.			ijerph19127332_makarova	0
+For the convenience of you and readers, we have made full descriptions of some acronyms, and the modifications are as follows: “Therefore, in this study, the Best Tracer Method (BTM) was employed. This method has been used before”, “the Hazard Quotients (HQ) values between 1 and 10 indicate likely damage to human health”.	2	1	The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3.			ijerph19127332_makarova	0
+Thank you for your suggestions. We have added the description of sampling site in the materials and methods. We have replied to your first suggestion on Materials and Methods for the specific content to be added.	2	1	Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue.			ijerph19127332_makarova	0
+We have added the values for carcinogenic risk and non-carcinogenic risks in the support information.	2	1	In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR.			ijerph19127332_makarova	0
+Thank you for your proposal. For this part, we have added to the methods section, and followed as: “The Hazard Quotients (HQ) is the ratio of daily intake dose of pollutants to reference dose, which is used to characterize the levels of human exposure to non-carcinogenic contaminants through a single pathway which represents the level of non‒carcinogenic risk”.	2	1	Page 3, lines 121-124:Please, add some information: How did you collect dust samples? How much soil (kg) did you collect for each sample?			ijerph19127332_makarova	0
+Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities”	2	1	As an example, they may analyze the influences of food intake and food intake on heavy metal exposure.			ijerph19127332_makarova	0
+Thank you for your advice. At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5). Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas. We have examined the tracer elements in the food. And when we calculated the SIR, we also used the parameter “Cfood” (equation 1) to analyze the food intake. In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR. Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue. At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd. The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas. Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Point 2.	2	1	It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5).			ijerph19127332_makarova	0
+Thanks for your advice, and this explanation has been removed from the manuscript, it is changed to “this method analyzes the concentration of tracer elements in the soil to which children are exposed, the children's intake of food, their excreted feces and urine, and the content of the tracer element in the children’s food, feces, and urine.”	2	1	Thus, we cannot provide the data for each day.			ijerph19127332_makarova	0
+; Shi, X.L. ; Wu, K.S. Human Body Burden of Heavy Metals and Health Consequences of Pb Exposure in Guiyu, an E-Waste Recycling Town in China. Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428” And we changed content as “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Point 3.	2	1	Specify the mixed standards and certified reference materials used for calculating accuracy of the digestion procedure.			ijerph19127332_makarova	0
+Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C). Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.	2	1	Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_makarova	0
+Thank you for your suggestion. We have added the content we did not mention in this part, but the soil was not quartered, so there is no supplement. Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills. And 20 g of soil and as much dust (5-20 g) as possible were collected during the sampling process.” Point 5.	2	1	Temperature of the heating plate for faces sample digestion.			ijerph19127332_makarova	0
+Dried samples (1 g) were digested to evaporate at low temperatures of 55°C on a heating plate with 3 mL concentrated nitric acid, 3 mL hydrogen fluoride, and 1 mL perchloric acid (HNO3‒HF‒HClO4). Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2). Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min. The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C. The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3. The conditions of microwave digestion are the same as mentioned above. Finally, the digest was analyzed by HR‒ICP‒MS. Dried soil and dust samples (0.5 g) were digested as same as feces and food samples on a heating plate by HNO3‒HF‒HClO4. And then samples were microwaved with HNO3‒H2O2. The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y. In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn. The standard curve of concentration was used to determine the sample concentration was established by heavy metal standard (all standards were from The Nonferrous Metals Society of China).” Point 4. Thanks for your advice! In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way. The digestion process is repeated until the sample becomes sticky.	2	1	This study estimated the soil ingestion of 66 17 children from e‒waste disassembly areas by collecting and analyzing selected tracer elements in 18 matched samples of their consumed food, feces, and urine, as well as soil samples from their play 19 areas.			ijerph19127332_makarova	0
+Thank you for your correction. We have added (a, b, c, ect) to Figure 1 and Figure S1,2,3,4. The title of Figure 1 has been changed to “Figure 1. Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h).” The title of Figure S1 has been changed to “Fig. S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”. The title of Figure S2 has been changed to “Fig. S2 Histogram and basic statistical parameters of daily food ingestion (a) (g/d, ww) and feces (b) (g/d, dw) and urine (c) (mL/d) excretion for investigated child population.”. The title of Figure S3 has been changed to “Fig. S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”. The title of Figure S4 has been changed to “Fig. S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.	2	1	The title of Figure 1 has been changed to “Figure 1.			ijerph19127332_makarova	0
+Thanks for your reminder. In our study, 66 children were taken part in our analysis. But we failed to get all children’s feces and urine sample every day. Thus, we collected sample number was less the involved 66 children.	2	1	The standard curve of concentration was used to determine the sample concentration was established by heavy metal standard (all standards were from The Nonferrous Metals Society of China).” Point 6.			ijerph19127332_perova	0
+Thank you for your reminder. We have uniformed the units to “(mL/d)” Point 4.	2	1	Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min.			ijerph19127332_perova	0
+At present, the related content in manuscript has been revised to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			ijerph19127332_perova	0
+In the manuscript, the data is with outlier. However, we used the data without outlier to construct the figure. The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 5. At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.	2	1	Title- ‘in’ should be used for larger areas, can you add the study locations?			ijerph19127332_perova	0
+We have reorganized the SM figures and the manuscript content. The modifications are as follows: “The frequency distribution histograms show few outliers, most being high values (Figure S3 (a) and Figure S4). Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers. Other outliers may be due to the behavior of certain children, such as pica behavior or spending unusually large amounts of time playing in grassland. The medians of the SIR values after removing the high values (see Figure S3(b) and Figure 1) were taken as the final SIR values for children living in the studied e‒waste dismantling site in South China.” Point 7.	2	1	I suggested the major revision before publication.			ijerph19127332_perova	0
+It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5). However, since negative SIR values are physically meaningless, the RI of the SIR for children living in e‒waste dismantling sites is 0‒730.4 mg/d. In this study, 95th per-centile values (383.3 mg/g) would be as recommended value.” Point 8.	2	1	Reviewer #1 Point 1、This study estimated the heavy metal exposure to children through soil and dust Ingestion in electronic waste (e-waste) dismantling areas.			ijerph19127332_perova	0
+It has been changed to “The 95th HQ values based on SIR mean value decreased in the order of Pb > As > Cr > Cu > Ni > Zn > Cd and all of them were below 1.” Point 9.	2	1	However, we used the data without outlier to construct the figure.			ijerph19127332_perova	0
+We have revised all the figure captions in SM.	2	1	Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers.			ijerph19127332_perova	0
+Thanks for your advice, we have made the changes to the title following:” Estimation of Children’s Soil/Dust Ingestion Rates and Health Risk at E‑Waste Dismantling Area”	2	1	The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas.			ijerph19127332_perova	0
+Thanks for your suggestion. We have introduced the sampling sites and added the following information: “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428 Point 3.	2	1	At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5).			ijerph19127332_perova	0
+Thanks for your suggestion, we have sorted out the method part again, and now the modification is as follows: “Feces samples and food samples were pretreated in the same way. The digestion process was repeated until the sample becomes sticky.	2	1	Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.			ijerph19127332_perova	0
+For this question, our answer is following: this data is from the Technical Guidelines for Soil Pollution Risk Assessment of Construction Land in China. The website address of the guidelines is https://www.mee.gov.cn/ywgz/fgbz/bz/bzwb/trhj/201912/W020191224560850148092.pdf Will like to see ethical approval for the project/research? Ethical approval from which institution? Were Consent/assent from participants obtained? For this problem, we conducted a questionnaire survey when collecting samples before the experiment, and all the guardians of the sample providers knew and agreed to the experiment.	2	1	Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_perova	0
+Collecting samples followed the USEPA recommendation of a period of 28 hours from food to feces and urine. Thus, we cannot provide the data for each day. And we also revised the table 1 legend (removing the “each day”).	2	1	Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			ijerph19127332_perova	0
+For the convenience of you and readers, we have made full descriptions of some acronyms, and the modifications are as follows: “Therefore, in this study, the Best Tracer Method (BTM) was employed. This method has been used before”, “the Hazard Quotients (HQ) values between 1 and 10 indicate likely damage to human health”.	2	1	It was only mentioned towards the end – Discussion Answer: Thanks for your suggestion.			ijerph19127332_perova	0
+Thank you for your suggestions. We have added the description of sampling site in the materials and methods. We have replied to your first suggestion on Materials and Methods for the specific content to be added.	2	1	Sample Preparation and Instrumental Analysis - Why were the food samples microwaved?			ijerph19127332_perova	0
+We have added the values for carcinogenic risk and non-carcinogenic risks in the support information.	2	1	And when we calculated the SIR, we also used the parameter “Cfood” (equation 1) to analyze the food intake.			ijerph19127332_perova	0
+Thank you for your proposal. For this part, we have added to the methods section, and followed as: “The Hazard Quotients (HQ) is the ratio of daily intake dose of pollutants to reference dose, which is used to characterize the levels of human exposure to non-carcinogenic contaminants through a single pathway which represents the level of non‒carcinogenic risk”.	2	1	Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_perova	0
+Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities”	2	1	In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn.			ijerph19127332_perova	0
+Thank you for your advice. At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5). Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas. We have examined the tracer elements in the food. And when we calculated the SIR, we also used the parameter “Cfood” (equation 1) to analyze the food intake. In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR. Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue. At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd. The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas. Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Point 2.	2	1	Please, use element symbols not the entire name in all the manuscript.			ijerph19127332_perova	0
+Thanks for your advice, and this explanation has been removed from the manuscript, it is changed to “this method analyzes the concentration of tracer elements in the soil to which children are exposed, the children's intake of food, their excreted feces and urine, and the content of the tracer element in the children’s food, feces, and urine.”	2	1	It has been changed to “The 95th HQ values based on SIR mean value decreased in the order of Pb > As > Cr > Cu > Ni > Zn > Cd and all of them were below 1.” Point 9. correct the figure caption in SM.			ijerph19127332_perova	0
+; Shi, X.L. ; Wu, K.S. Human Body Burden of Heavy Metals and Health Consequences of Pb Exposure in Guiyu, an E-Waste Recycling Town in China. Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L. International Journal of Environmental Research and Public Health, 2021, 18(23):12428. https://doi.org/10.3390/ijerph182312428” And we changed content as “The e-waste dismantling area we studied is an E-Waste Recycling Town located in South China, where the possible human body burden and health consequences of heavy metals exposure have been reported [30].” Point 3.	2	1	Introduction 30 The potentially harmful environmental and human health effects of primitive elec- 31 tronic‒waste (e‒waste) recycling processes, including manual disassembly, roasting, acid 32 leaching, and open burning, have caused concern around the world, particularly in rap- 33 idly industrializing and urbanizing developing countries such as China, India, and Vi- 34 etnam [1,2].			ijerph19127332_perova	0
+Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C). Then, the feces were freeze‒dried (under vacuum conditions, the vacuum freeze drier temperature is ‒40°C to ‒50°C for 48 h) after measuring the weight with a vacuum freeze dryer.” Point 4.	2	1	Dear Authors and Editor, the work presents interesting results on estimation of the soil ingestion of selected children from e‒waste disassembly areas by collecting and analyzing selected tracer metals in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas.			ijerph19127332_perova	0
+Thank you for your suggestion. We have added the content we did not mention in this part, but the soil was not quartered, so there is no supplement. Now the modification is as follows: “Collect dust indoors or outdoors by cleaning dust from areas such as tables and windowsills. And 20 g of soil and as much dust (5-20 g) as possible were collected during the sampling process.” Point 5.	2	1	The modifications are as follows: “The frequency distribution histograms show few outliers, most being high values (Figure S3 (a) and Figure S4).			ijerph19127332_perova	0
+Dried samples (1 g) were digested to evaporate at low temperatures of 55°C on a heating plate with 3 mL concentrated nitric acid, 3 mL hydrogen fluoride, and 1 mL perchloric acid (HNO3‒HF‒HClO4). Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2). Digestion was performed at 120°C for 5 min, then 160°C for 5 min, and finally 180°C for 15 min. The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C. The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3. The conditions of microwave digestion are the same as mentioned above. Finally, the digest was analyzed by HR‒ICP‒MS. Dried soil and dust samples (0.5 g) were digested as same as feces and food samples on a heating plate by HNO3‒HF‒HClO4. And then samples were microwaved with HNO3‒H2O2. The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y. In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn. The standard curve of concentration was used to determine the sample concentration was established by heavy metal standard (all standards were from The Nonferrous Metals Society of China).” Point 4. Thanks for your advice! In view of the imperfection of the method and the defects of expression, we have added and improved this part, and the modification is as follows: “Feces samples and food samples were pre-treated in the same way. The digestion process is repeated until the sample becomes sticky.	2	1	Introduction 30 The potentially harmful environmental and human health effects of primitive elec- 31 tronic‒waste (e‒waste) recycling processes, including manual disassembly, roasting, acid 32 leaching, and open burning, have caused concern around the world, particularly in rap- 33 idly industrializing and urbanizing developing countries such as China, India, and Vi- 34 etnam [1,2].			ijerph19127332_perova	0
+Thank you for your correction. We have added (a, b, c, ect) to Figure 1 and Figure S1,2,3,4. The title of Figure 1 has been changed to “Figure 1. Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h).” The title of Figure S1 has been changed to “Fig. S1 Histogram and basic statistical parameters of investigated child population age (a), height (b), and weight (c)”. The title of Figure S2 has been changed to “Fig. S2 Histogram and basic statistical parameters of daily food ingestion (a) (g/d, ww) and feces (b) (g/d, dw) and urine (c) (mL/d) excretion for investigated child population.”. The title of Figure S3 has been changed to “Fig. S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”. The title of Figure S4 has been changed to “Fig. S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.	2	1	I suggested the major revision before publication.			ijerph19127332_perova	0
+Thank you for this suggestion. While it would have been interesting to explore this further, our study focuses on retrospectively assessing and comparing disease progression within subgroups of patients receiving TOC. Patients not receiving TOC would have likely had a different presentation, progression and their follow-up would be slightly out of the scope of our observational retrospective study. When we compared patients who received TOC with the 86 severe patients who did not receive TOC (excluded from this analysis), the TOC group was far more severe, so there is no control group that could be matched. In addition, these patients did not have control CT scans.	2	1	1 Response to Reviewer 2 Comments Point 1: 2.			jcm11051247_makarova	0
+Following this suggestion, we have included in the analysis a calculation according to the FiO2. This has a cut-off which correlates to the oxygen flow cut-off.	2	1	The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).			jcm11051247_makarova	0
+This is an interesting point. While knowledge of variants is of great importance from an epidemiologic point of view as they have different transmission rates and can cause diseases of different severity, the mechanism of the acute inflammatory response in severe infection seems to be similar regardless the strain. While we did not perform specific sequencing of the strain for every patient as this wouldn’t have been feasible, patients were included before the Delta variant emerged and we expect most of them to have had the initial D614G and UK B.1.1.7. Nonetheless, we have included a comment addressing this aspect in our limitations section to highlight this area of uncertainty.	2	1	Point 2: The proposed timing of Tocilizumab administration based on oxygen flow rate is interesting but undoubtedly requires a more detailed statistical analysis to convince the readers.			jcm11051247_makarova	0
+We agree with the referee that the reasons for the exclusions were unclear and have rephrased accordingly. Addresed in text: „All those excluded were not administered TOC either because of contraindications or TOC supply gaps.” Point 3: 3.	2	1	1 Response to Reviewer 2 Comments Point 1: 2.			jcm11051247_makarova	0
+We added the data using FiO2 ROC curve. The area under the curve for FiO2 ROC curve was smaller than the area under the curve for oxygen flow. In addition, the cut-off value for FiO2 corresponded to the cut-off for the oxygen flow rate, so the two groups in Table 3 would be similar.	2	1	1 Response to Reviewer 1 Comments Point 1: This study has a design flaw.			jcm11051247_makarova	0
+We added this information also in Table 3. We analysed FiO2 (depending on the method of oxygen delivery - nasal canulae, HNF, MV), as you suggested and the cut-off value of FiO2 corresponded to the cut-off for the oxygen flow rate, so the two groups in Table 3 would be similar.	2	1	The impact of Tocilizumab on radiological changes can not be adequately analyzed without a control group not receiving the drug.			jcm11051247_makarova	0
+Addressed in text and in Table 3.	2	1	Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.			jcm11051247_makarova	0
+Thank you for pointing this out, we have changed Table 1 accordingly and we removed the imaging data, which as you mentioned, was the same as “before TOC” in Table 2.	2	1	The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).			jcm11051247_makarova	0
+Addressed, see previous comments and answers.	2	1	Results 2 The oxygen flow rate of 13 L/min measured by ROC as a cutoff value for TOC administration should be presented by FiO2.			jcm11051247_makarova	0
+separation of sections and subheadings, revising all the text, adding paragraphs & references, adding/changing Tables & Figures): 1) Abstract was revised.	2	1	Some specific comments: The abstract could be more concise.			land11030363_makarova	0
+Sorry for the technical error. The sample set was divided into two subsets (75% and 25%) for calibration (86 samples) and validation (28 samples).	2	1	Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+The sections were separated as recommended.	2	1	The abstract and conclusion parts lack conciseness and prominence.			land11030363_makarova	0
+The comment was fully considered: the PCA and partial correlation analysis was adopted form the recommended study (e.g., Cheng et al., 2019). Correspondingly, separate subheadings (with new Table 4 and Figure 4) were added. Generally it was considered in Introduction and Materials & Methods, Results and Discussion sections. Such approach helped to analyze the underlying mechanisms.	2	1	Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			land11030363_makarova	0
+It was corrected to “modeling”.	2	1	Is the number of samples sufficient for the purpose of the research?			land11030363_makarova	0
+It was corrected. pH was measured in water solution. (Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?	2	1	Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.			land11030363_makarova	0
+It was revised as recommended. (Line: 244-249) Line 174: “…resulting in 12 spectra”.	2	1	Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			land11030363_makarova	0
+The text was revised. Each single measurement was based on internally averaged of 50 spectra (3 times). Each sample was measured from four positions (90°rotating the Petri dish each time). Consequently, 4 rotation x 3 time = 12 spectra were obtained. It is one of the commonly used method and several references are available. Line;252-259 Line 176: “…resulted 50 spectra…”.	2	1	Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			land11030363_makarova	0
+See previous comment.	2	1	is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			land11030363_makarova	0
+The whole paragraph was revised. Line: 262-269. We would like to note that in practice a large variety of pre-processing techniques are used with different gap and moving window size to improve prediction quality (Luce et al. 2017, Mammadov at al. 2020). Yet, the quality of prediction still depends on several factors (measurement method, used device etc.).	2	1	7) Conclusions was modified to show its practical outcomes for the local and international readers.			land11030363_makarova	0
+The text was revised, see previous comment.	2	1	Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources.			land11030363_makarova	0
+It was revised as “change in pH, temperature, precipitation by elevation”. Line:334-337.	2	1	It is not clear how the 12 spectra were obtained here.			land11030363_makarova	0
+Corrected to “soil type”.	2	1	Too many parentheses are used in the essay, which is not recommended in writing.			land11030363_makarova	0
+Both methods are used to characterize relations between soil properties and spectra. Based on our experience, for current study (or similar studies) relations among soil properties, and between PC1 and soil properties are well characterized by Spearman, and Pearson is more relevant to the relations between spectra and soil properties.	2	1	Response to Reviewer ≠1 comment Many thanks for your response.			land11030363_makarova	0
+Line:515-518.	2	1	"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			land11030363_makarova	0
+Line:43, 440 Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here.	2	1	Readers may be more interested in your analysis and interpretation.			land11030363_makarova	0
+It was modified. Line: 438-441 Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	2	1	The abstract and conclusion parts lack conciseness and prominence.			land11030363_makarova	0
+Line: 696-699 Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	2	1	Line 264: “…CaCO3 and Fe),” Where is the left parenthesis?			land11030363_makarova	0
+Line:702-705 Line 409: Note the singular and plural in the sentence.	2	1	In addition, it needs support with more and more up-to-date recent articles.			land11030363_makarova	0
+Corrected to “CaCO3”. Line: 750-752 Line 455: “soil mineralogy”?	2	1	Please pay attention to the singular and plural in the sentence.			land11030363_makarova	0
+It was replaced with “clay mineralogy”.	2	1	Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_makarova	0
+Line 736-740 Line 471, 480, 520: “(r = 0.4*)” “(r =-0.52*)” “(r = 0.40*)”, etc.	2	1	Note the singular and plural in the sentence.			land11030363_makarova	0
+Line:759-762 Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what?	2	1	prediction mechanism, prediction accuracy, etc.)			land11030363_makarova	0
+It was revised. It was related to the contribution of the basic soil properties (e.g. controlling factor CaCO3, Fe, clay) to the prediction of M3 extractable elements Line:780-786.	2	1	Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit.			land11030363_makarova	0
+Corrected or revised. Line:786-789 Line 523: “correspond absorption” should be “correspond to absorption”.	2	1	Author Response Please see the attachment Author Response File: Author Response.docx			land11030363_makarova	0
+It was edited. Line:790-791 Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence?	2	1	Specific comments: When the samples are grouped, it is generally carried out according to 7:3 or 3:1 or a certain ratio.			land11030363_makarova	0
+Partial correlation analysis showed that the contribution of Fe was more important than that of SOC though the moderate correlation existed between them. Line: 786-791 Lines 541-543: Is this sentence appropriate here?	2	1	Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			land11030363_makarova	0
+We have followed the instruction. The change was marked with the red letter.	4	1	The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.			land11030363_makarova	0
+The abstract was revised to show all key results.	4	1	Too many parentheses are used in the essay, which is not recommended in writing.			land11030363_makarova	0
+Introduction was modified.	4	1	Line 135: “impairment water resources” Is this expression appropriate?			land11030363_makarova	0
+The manuscript was checked for missing articles and edited.	4	1	Line 264: “…, and hence parent material.” It’s not clear here.			land11030363_makarova	0
+We reduced the number of parentheses significantly.	4	1	c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned.			land11030363_makarova	0
+It was removed.	4	1	The introduction part can highlight the key points and innovations of this paper.			land11030363_makarova	0
+Soory for the technical error. In the previous version, pH values were related to the determination in KCL solution. It was replaced with pH values in H2O water.	4	1	Line 233: It’s usually written as “Results and discussion”.			land11030363_makarova	0
+It was edited both in the text and tables.	4	1	"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			land11030363_makarova	0
+It was corrected.	4	1	c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned.			land11030363_makarova	0
+Thanks. The conclusions were rewritten to reflect the key results.	4	1	What does the controlled laboratory environment you describe look like?			land11030363_makarova	0
+We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. New subheading (3.2. Land use effect on soil properties) was added. New Table 4 and Figure 4 were added. Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear). We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of section and subheadings, revising all the text, adding paragraphs and references, adding or changing Tables and Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. a) Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. b) New subheading (3.2. Land use effect on soil properties) was added. It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1. c). New Table 4 and Figure 4 were added. Table 4 is related to partial correlation analysis with consideration of soil properties, and Figure 4 is related to the principal component analysis (biplot). d) Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 6) Discussion with new 3 subheading was significantly expanded and linked to the references and the novelty was highlighted. 7) Conclusions of the study was modified to show its practical outcomes for the local and international readers. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Due to the separating the Results and Discussion section, overlapping of some comments made by the reviewers and associated changes modified all the text of the manuscript was modified. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear).	2	1	Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			land11030363_makarova	0
+The section was revised and information regarding solution standards and detection limit was added. (M&M. subheading 2.2). Chemical analyses and descriptive statistics of the results b) It is necessary to explain the method of sampling using the appropriate literature.	2	1	6) Discussion with new 3 subheading was significantly expanded and linked to the references, and the novelty was highlighted.			land11030363_makarova	0
+The sampling scheme was explained as recommended (M&M, subheading 2.1). In general, sampling scale changes between 1:12 000 and 1:250 000 (Soil Survey Staff, USDA NRCS 2016). In this pioneering study area, soil properties are highly variable attributed to topography, land use history and land tenure ownerships (mainly 0.2-0.3 hectares and fragmented shrubbery areas) and geological setting that makes difficult to apply regular sampling scheme. Therefore, the sampling locations were randomly (irregularly) designated, yet cover variations in land use, topography, geological substrate and the erosive state of soil continuum thereby resulted in 114 samples from 525 ha used under four land use types (1 sample per ~ 5 hectares).	2	1	What does the controlled laboratory environment you describe look like?			land11030363_makarova	0
+Two references were added. (Subheading 2.1) The “Conclusions section” is not very successful concerning the presentation of results.	2	1	The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+As mentioned above, we restructured the introduction section and included new paragraphs to illuminate the importance of the study. Special focus was given to clarifying prediction mechanisms for the studied soil properties and land use effect on soil properties.	2	1	What does the controlled laboratory environment you describe look like?			land11030363_makarova	0
+The suggested study was reviewed and included to the reference list. It helped us to explain the results of our study (e.g. prediction of Fe and Cd using basic soil properties, clay mineralogy and (soil type) and micro-nutrients).	2	1	Line 233: It’s usually written as “Results and discussion”.			land11030363_makarova	0
+Response: We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of section and subheadings, revising all the text, adding paragraphs and references, adding or changing Tables and Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. a) Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. b) New subheading (3.2. Land use effect on soil properties) was added. It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1. c). New Table 4 and Figure 4 were added. Table 4 is related to partial correlation analysis with consideration of soil properties, and Figure 4 is related to the principal component analysis (biplot). d) Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 6) Discussion with new 3 subheading was significantly expanded and linked to the references and the novelty was highlighted. 7) Conclusions of the study was modified to show its practical outcomes for the local and international readers. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Due to the separating the Results and Discussion section, overlapping of some comments made by the reviewers and associated changes modified all the text of the manuscript was modified. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript. Two version of the manuscript was uploaded: edited main manuscript (with track change), and edited main manuscript (clear).	2	1	(Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?			land11030363_perova	0
+Sorry for the technical error. The sample set was divided into two subsets (75% and 25%) for calibration (86 samples) and validation (28 samples).	2	1	Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			land11030363_perova	0
+The sections were separated as recommended.	2	1	(Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?			land11030363_perova	0
+The comment was fully considered: the PCA and partial correlation analysis was adopted form the recommended study (e.g., Cheng et al., 2019). Correspondingly, separate subheadings (with new Table 4 and Figure 4) were added. Generally it was considered in Introduction and Materials & Methods, Results and Discussion sections. Such approach helped to analyze the underlying mechanisms.	2	1	Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			land11030363_perova	0
+It was corrected	2	1	Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_perova	0
+pH was measured in water solution.	2	1	The definite article “the” is missing in many places in the whole article.			land11030363_perova	0
+It was revised as recommended.	2	1	Modified content should be marked in the revised manuscript for easy review by reviewers.			land11030363_perova	0
+The text was revised. Each single measurement was based on internally averaged of 50 spectra (3 times). Each sample was measured from four positions (90°rotating the Petri dish each time). Consequently, 4 rotation x 3 time = 12 spectra were obtained. It is one of the commonly used method and several references are available.	2	1	Line 520: The correlation between K content and P content is not very high (r = 0.40*), but their corresponding VIP patterns are “identical”, how do you explain it?			land11030363_perova	0
+See previous comment.	2	1	Line 233: It’s usually written as “Results and discussion”.			land11030363_perova	0
+The whole paragraph was revised. Line: 262-269. We would like to note that in practice a large variety of pre-processing techniques are used with different gap and moving window size to improve prediction quality (Luce et al. 2017, Mammadov at al. 2020). Yet, the quality of prediction still depends on several factors (measurement method, used device etc.).	2	1	Line 104, 195, 223, 544: Is it “modelling” or “modeling”?			land11030363_perova	0
+The text was revised, see previous comment.	2	1	Response to Reviewer ≠1 comment Many thanks for your response.			land11030363_perova	0
+It was revised as “change in pH, temperature, precipitation by elevation”. Line:334-337.	2	1	The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.			land11030363_perova	0
+Corrected to “soil type”.	2	1	Attention should be paid to the number of decimal places in all numbers throughout the text.			land11030363_perova	0
+Both methods are used to characterize relations between soil properties and spectra. Based on our experience, for current study (or similar studies) relations among soil properties, and between PC1 and soil properties are well characterized by Spearman, and Pearson is more relevant to the relations between spectra and soil properties.	2	1	Line 135: “impairment water resources” Is this expression appropriate?			land11030363_perova	0
+Line:515-518.	2	1	Table 1: The pH values are different from those in the previous version.			land11030363_perova	0
+Line:43, 440 Line 354-355: “…, in our case 11 bands…” “…in our case 10 wavebands…” It's confusing here.	2	1	Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			land11030363_perova	0
+It was modified. Line: 438-441 Lines 356-357: Can the first derivative preprocessing method be used to remove illumination differences?	2	1	Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources.			land11030363_perova	0
+Line: 696-699 Lines 360-362: “… due to overtones and combination of fundamental vibrations of soil organic matter occurring in the VIS and NIR region.” The meaning of this sentence is not expressed clearly.	2	1	What does the controlled laboratory environment you describe look like?			land11030363_perova	0
+Line:702-705 Line 409: Note the singular and plural in the sentence.	2	1	The introduction part can highlight the key points and innovations of this paper.			land11030363_perova	0
+Corrected to “CaCO3”. Line: 750-752 Line 455: “soil mineralogy”?	2	1	Please pay attention to the singular and plural in the sentence.			land11030363_perova	0
+It was replaced with “clay mineralogy”.	2	1	Reference to formal analytical methods must be made in a scientific manner.			land11030363_perova	0
+Line 736-740 Line 471, 480, 520: “(r = 0.4*)” “(r =-0.52*)” “(r = 0.40*)”, etc.	2	1	Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			land11030363_perova	0
+Line:759-762 Lines 516-518: You call attention to the spectral signature at 2326 nm, and then what?	2	1	Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit.			land11030363_perova	0
+It was revised. It was related to the contribution of the basic soil properties (e.g. controlling factor CaCO3, Fe, clay) to the prediction of M3 extractable elements Line:780-786.	2	1	Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources.			land11030363_perova	0
+Corrected or revised. Line:786-789 Line 523: “correspond absorption” should be “correspond to absorption”.	2	1	In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are.			land11030363_perova	0
+It was edited. Line:790-791 Lines 522-525: Is there a necessary connection between the first half of the sentence and the second half of the sentence?	2	1	However, there are some important issues that need to be addressed:			land11030363_perova	0
+Partial correlation analysis showed that the contribution of Fe was more important than that of SOC though the moderate correlation existed between them. Line: 786-791 Lines 541-543: Is this sentence appropriate here?	2	1	Author Response Please see the attachment Author Response File: Author Response.docx			land11030363_perova	0
+edited	2	1	Lines 456-457: What does this sentence do here?			land11030363_perova	0
+The abstract was revised to show all key results.	4	1	It is not clear how the 12 spectra were obtained here.			land11030363_perova	0
+Introduction was modified.	4	1	However, there are some issues that need to be resolved before this manuscript can be considered for publication.			land11030363_perova	0
+The manuscript was checked for missing articles and edited.	4	1	As far as I know, most articles use only one moving window size.			land11030363_perova	0
+We reduced the number of parentheses significantly.	4	1	The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			land11030363_perova	0
+It was removed.	4	1	Is the number of samples sufficient for the purpose of the research?			land11030363_perova	0
+Soory for the technical error. In the previous version, pH values were related to the determination in KCL solution. It was replaced with pH values in H2O water.	4	1	It would be wise to give more international flavor in the sections of Introduction and Discussion.			land11030363_perova	0
+It was edited both in the text and tables.	4	1	It undoubtedly increases your experimental workload, and you do not explain it later.			land11030363_perova	0
+It was corrected.	4	1	It undoubtedly increases your experimental workload, and you do not explain it later.			land11030363_perova	0
+Thanks. The conclusions were rewritten to reflect the key results.	4	1	"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			land11030363_perova	0
+We would like to thank you for your valuable comments and directions. We have carefully considered all issues and have revised the manuscript completely (e.g. separation of sections and subheadings, revising all the text, adding paragraphs & references, adding/changing Tables & Figures): 1) Abstract was revised. 2) Introduction was revised, and 2 new paragraphs were added. Such modification leaded to the clear description of the objectives. 3) Material and Method section was revised. Revised version contains 5 subheading. 4) The results and discussion sections were separated, and their subheadings were reorganized. In the revised manuscript the Results contain 5 subheading and Discussion contain 3subheading. 5) Presentation of the results are improved with consideration of reviewers’ comments. Original Manuscript contained 3 Tables and 3 Figures. Revised manuscript contains 4 Tables and 5 Figures and 1 supplementary table and 1 supplementary figure. New subheading (3.2. Land use effect on soil properties) was added. New Table 4 and Figure 4 were added. Clarification: Table 1 and Table 3 were not changed. Old Table 2 is new Figure 2. Old Figure 2 is new Figure 3. Old Figure 3 is new Figure 5. 8) New 12 references were added to update the manuscript to address reviewers’ comments. Thus, to avoid possible confusion we provided the location (subheading or line) for any change introduced. Our point-to-point response to the comments of the reviewers is detailed below. Subheading or line numbers refer to the line numbers in the revised manuscript.	2	1	However, there are some important issues that need to be addressed:			land11030363_perova	0
+The section was revised and information regarding solution standards and detection limit was added. (M&M. subheading 2.2). Chemical analyses and descriptive statistics of the results b) It is necessary to explain the method of sampling using the appropriate literature.	2	1	Line;252-259 Line 176: “…resulted 50 spectra…”.			land11030363_perova	0
+The sampling scheme was explained as recommended (M&M, subheading 2.1). In general, sampling scale changes between 1:12 000 and 1:250 000 (Soil Survey Staff, USDA NRCS 2016). In this pioneering study area, soil properties are highly variable attributed to topography, land use history and land tenure ownerships (mainly 0.2-0.3 hectares and fragmented shrubbery areas) and geological setting that makes difficult to apply regular sampling scheme. Therefore, the sampling locations were randomly (irregularly) designated, yet cover variations in land use, topography, geological substrate and the erosive state of soil continuum thereby resulted in 114 samples from 525 ha used under four land use types (1 sample per ~ 5 hectares).	2	1	It is also not clear how 50 spectra were obtained.			land11030363_perova	0
+Two references were added. (Subheading 2.1) The “Conclusions section” is not very successful concerning the presentation of results.	2	1	Is the number of samples sufficient for the purpose of the research?			land11030363_perova	0
+As mentioned above, we restructured the introduction section and included new paragraphs to illuminate the importance of the study. Special focus was given to clarifying prediction mechanisms for the studied soil properties and land use effect on soil properties.	2	1	Lines 325-328: “Regardless of… spectra.” Is there a necessary connection between these two sentences?			land11030363_perova	0
+The suggested study was reviewed and included to the reference list. It helped us to explain the results of our study (e.g. prediction of Fe and Cd using basic soil properties, clay mineralogy and (soil type) and micro-nutrients).	2	1	It is not clear how the 12 spectra were obtained here.			land11030363_perova	0
+We agree, and we have noted this point in the revised study limitations section.	2	1	I know, there were 7 missing values within the 310.			life12060901_makarova	0
+Interesting point, which we have added to the revised Conclusion and Implications section.	2	1	The authors did a cross-sectional online survey with a total sample of 1649 adults across four countries (Norway, UK, USA, and Australia).			life12060901_makarova	0
+Thank you for the kind response.	2	1	Hence, I have some comments/suggestions that I hope will help the authors to further develop this line of work: Authors’ response: Thank you for pointing out these mistakes, and we trust the editorial office will assist us further with any remaining details in the final stage of the process.			life12060901_makarova	0
+Please see our comments to each of the listed issues below.	2	1	Please use a consistent writing style throughout the whole manuscript.			life12060901_makarova	0
+The first mentioned statement has been removed altogether in the revised manuscript. The second statement has been modified in accordance with the reviewer’s view (see revised section 4.3).	2	1	Authors’ response: Table 1 has been revised accordingly.			life12060901_makarova	0
+Thank you for pointing this out. We have removed the relevant section in accordance with this guidance.	2	1	People would get really irritated and discouraged by such a situation.			life12060901_makarova	0
+Thank you for pointing this out, we have revised the sentence (see section 4.1).	2	1	Authors’ response: Thank you for the kind – and quick – response!			life12060901_makarova	0
+We have revised the sentence; see section 4.4.	2	1	In my opinion, this paper deals with a very important and timely topic and is well structured.			life12060901_makarova	0
+We have revised the sentence; see section 5.	2	1	Otherwise, this is misleading the readership.			life12060901_makarova	0
+Thank you for pointing out these mistakes, and we trust the editorial office will assist us further with any remaining details in the final stage of the process.	2	1	In my opinion, this paper deals with a very important and timely topic and is well structured.			life12060901_makarova	0
+Alignment has been fixed.	2	1	The introduction guides the reader smoothly into this topic, all necessary background information (i.e., citing appropriate literature) is given.			life12060901_makarova	0
+Table 1 has been revised accordingly.	2	1	Authors’ response: Thank you for the kind words.			life12060901_makarova	0
+Italics have been removed, see revised section 2.3.	2	1	People who have infected with COVID-19 sometimes report long lasting COVID symptoms like the loss of taste and smell.			life12060901_makarova	0
+We are unsure how this problem occurred. We have fixed the problems in the revised tables.	2	1	The authors of this paper examined whether sociodemographic as well as health-related differences between COVID-19 patients with and without long COVID could be observed.			life12060901_makarova	0
+Thank you for pointing out, we have addressed this issue throughout the revised manuscript.	2	1	People who have infected with COVID-19 sometimes report long lasting COVID symptoms like the loss of taste and smell.			life12060901_makarova	0
+We have corrected the two tables according to this guidance.	4	1	However, what I disliked was the interpretation and discussion of the empirical results.			life12060901_makarova	0
+Thank you for noticing. We have corrected the manuscript according to this guidance.	4	1	Besides these two major issues, I detected some smaller errors and miss-spellings (note, this list in not complete).			life12060901_makarova	0
+We agree, and we have noted this point in the revised study limitations section.	2	1	Authors’ response: Thank you for pointing this out.			life12060901_perova	0
+Interesting point, which we have added to the revised Conclusion and Implications section.	2	1	Authors’ response: Thank you for the kind – and quick – response!			life12060901_perova	0
+Thank you for the kind response.	2	1	People who have infected with COVID-19 sometimes report long lasting COVID symptoms like the loss of taste and smell.			life12060901_perova	0
+The first mentioned statement has been removed altogether in the revised manuscript. The second statement has been modified in accordance with the reviewer’s view (see revised section 4.3).	2	1	However, this table is still wrong, it must be 310 and not 303.			life12060901_perova	0
+Thank you for pointing this out. We have removed the relevant section in accordance with this guidance.	2	1	Author Response Reviewer 3 (R3): I thank the authors for submitting a revised version of their manuscript entitled “Self-reported long COVID in the general population: sociodemographic and health correlates in a cross-national sample”.			life12060901_perova	0
+Thank you for pointing this out, we have revised the sentence (see section 4.1).	2	1	I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential.			life12060901_perova	0
+We have revised the sentence; see section 4.4.	2	1	Otherwise, this is misleading the readership.			life12060901_perova	0
+We have revised the sentence; see section 5.	2	1	"R3: Wrong use of abbreviations: The authors introduced the abbreviation “GHQ-12” for the 12-item General Health Questionnaire (see page 4, line 161), but wrote in the text only ""GHQ"" (e.g., page 5, line 207; page 6, line 256; page 7, line 274)."			life12060901_perova	0
+Alignment has been fixed.	2	1	Please use a consistent writing style throughout the whole manuscript.			life12060901_perova	0
+Table 1 has been revised accordingly.	2	1	R3: However, I see several issues with the ending of this paper.			life12060901_perova	0
+Italics have been removed, see revised section 2.3.	2	1	I know, there were 7 missing values within the 310.			life12060901_perova	0
+We are unsure how this problem occurred. We have fixed the problems in the revised tables.	2	1	The reviewed paper deals with the COVID pandemic, especially with the “long COVID” phenomenon.			life12060901_perova	0
+Thank you for pointing out, we have addressed this issue throughout the revised manuscript.	2	1	All these statements are not true, only for two out of three outcomes, namely psychological distress and fatigue, but not for perceived stress (see also Figure 3).			life12060901_perova	0
+We have corrected the two tables according to this guidance.	4	1	Authors’ response: Table 1 has been revised accordingly.			life12060901_perova	0
+Thank you for noticing. We have corrected the manuscript according to this guidance.	4	1	***************************************************************************** Reviewer 1 (R1):			life12060901_perova	0
+In the paper we justify (at some length) the bending modulus we used, which we extrapolated from various sources, and explain why we believe the value in Boal and Ng’s study is likely too low. The reviewer did not question the logic or the sources used to arrive at this value, but nevertheless seemed surprised at the result. The reviewer claims that the parameter falls outside of the physical regime, but doesn’t offer a justification other than that the implied persistence length “seems too large”. However, the large persistence length simply implies that the cyanobacteria are stiff enough that they are largely unaffected by thermal fluctuations. Despite this, the trichomes in our simulations are still quite flexible in practice, as can be seen in Figure 4. Even if the virtual trichomes are somewhat too stiff, we do not feel that this would have a major impact on the results.	2	1	While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			life4030433_makarova	0
+We added the following statement to the end of Section 3.3: “The simulation results of the previous section for β > 0.5 are unlikely to be affected by the domain size, however, since the features in those simulations are on a much smaller scale than the domain size and are also more chaotic, as seen in the correlation length and the global alignment (Figure 5a,d).”	2	1	Qualitative comparisons are made to the fossil record and modern experiments.			life4030433_makarova	0
+We added the following clarification to the end of the non-dimensionalization section: “In the subsequent sections, simulations for β = 0 use a modified interaction force such that there is no cohesion, but the hard core repulsion is maintained, i.e., 0 0 c F = for 0 h ≥ , otherwise 0 c c F F = with β = 0.125.” (4)	2	1	Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_makarova	0
+We added two references to justify using the Lennard–Jones potential.	2	1	The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_makarova	0
+We added the following clarification: “This is simulated by generating a pseudo-random number x following a uniform distribution X ~ U(0,1), and reversing the gliding direction if x < ω⋅∆t.	2	1	The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_makarova	0
+We amended the following sentences of the discussion: “These differences may be due to the fact that we use a shallow domain (7.5 microns) to reduce the simulation run time, which may cause the pattern to be “squashed” as the domain ceiling prevents ridges from growing vertically” “Restricting the virtual trichomes such that they may only glide when in contact with the substratum or another trichome may promote further aggregation of the trichomes from streams into ridges, as a trichome would be less likely to successfully break away from a stream because it would lose much of its propulsive force as it lost contact with neighboring trichomes and/or the substratum.”	2	1	The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display.			life4030433_makarova	0
+We added the following sentence to Section 2.4: “Gliding requires contact with some (semi-)solid substrate in order to provide a reaction force to the gliding mechanism. We assume that the trichomes are immersed in highly viscous medium that allows them to glide freely in all directions. This medium could consist of the EPS the trichomes produce copiusly when gliding [Hoiczyk2000].” (8)	2	1	Qualitative comparisons are made to the fossil record and modern experiments.			life4030433_makarova	0
+The exact force is now given.	2	1	They also make quantitative measurements to describe the behaviour of the model trichomes.			life4030433_makarova	0
+The bibliography now has volume and page numbers.	2	1	Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_makarova	0
+The reviewer did not provide any examples where he/she felt a reference was warranted. We cited 55 sources, which we feel offers reasonable background and justification for our assumptions.	2	1	In response to the reviewers’ comments: (1) We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low.			life4030433_makarova	0
+References to the movies were added to the results section.	2	1	Qualitative comparisons are made to the fossil record and modern experiments.			life4030433_makarova	0
+We added the following sentence to the end of Section 2.8: “This value corresponds to a persistence length of 0.49 m, which implies that the trichomes would be practically unaffected by thermal fluctuations.”	4	1	The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_makarova	0
+We added the following paragraph to the discussion: “Many of the model parameters, such as trichome length, diameter, gliding speed, reversal frequency, etc. are easily measured and are well known. However, the bending modulus is more difficult to measure directly. We attempted to use the bending modulus implied from the relation α= kBT ε and Boal and Ng’s (2010, p. 4625) measurements of the trichome persistence length, but the resulting value seemed too low compared to measurements of other bacteria and in practice the virtual trichomes appeared flaccid during simulations. It is possible that the flexure seen in Boal and Ng’s trichomes was due more to the motility of the trichomes than random thermal fluctuations, in which case the above relation would no longer be valid, and a more complex model would be required to associate the observed geometry of the trichomes to their bending modulus. For example, Wolgemuth (2005) used an elastic model to estimate the bending modulus of M. xanthus by fitting the model to the flailing motions of a Myxobacterium stuck at one end.”	4	1	The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_makarova	0
+The symbol used to denote trichome length was changed to an uppercase lambda. The , ⋅⋅ is often used to denote a tuple in computer science, in this case a pair. The superscripts should have been subscripts. The notation used in Equations (5)–(8) was adopted from Bergou et al’s paper. h-hat is in fact a unit vector parallel to h, and we’ve clarified the text to this point.	2	1	Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_makarova	0
+We added a few sentences to the introduction that expand a bit on similar structures in the Petroff and Walter references.	2	1	In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			life4030433_makarova	0
+Actually, as Reviewer 1 noted, we use a viscosity that is 1000 times greater than that of water (following Wolgemuth et al. 2005), for the very reasons the reviewer mentioned.	2	1	They also make quantitative measurements to describe the behaviour of the model trichomes.			life4030433_makarova	0
+The statement has been clarified and is now: “The sum of the interaction’s opposing forces and torques is zero, thereby ensuring Newton's third law is respected.”	2	1	Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			life4030433_makarova	0
+Our estimate is simplistic, however we were unable to find a reliable measurement of this parameter in the literature, and so we resorted to some simple extrapolations.	2	1	In response to the reviewers’ comments: (1) We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low.			life4030433_makarova	0
+There are indeed additional parameters. We have added a complete list to Section 2.8. Many are explored to some extent in the results (beta, reversal frequency, domain dimensions, trichome R7 density), whereas most of the physical characteristics of the trichomes are known from published sources (diameter, length, gliding speed, bending modulus), and finally N was simply chosen so that trichomes had a “smooth” appearance.	2	1	In response to the reviewers’ comments: (1) We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low.			life4030433_makarova	0
+To avoid confusion on this point we have replaced this statement with the following: “We find that these errors are infrequent and acceptable as a trade-off for increased computational performance.” We have also added a comment to Section 2.5 regarding the Lennard–Jones interaction: “The attractive force has a short range, and is practically zero at h = 2θ.”	2	1	Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			life4030433_makarova	0
+It would be interesting to explore the approach the reviewer suggested; we have not yet explored more coarse grained models.	2	1	While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			life4030433_makarova	0
+Shepard et al. claim that the formation of the reticulate pattern occurs on a faster scale than cell growth and division, and so we did not consider this in our model. Including this might make the results more robust, however, and we added a comment to the discussion.	2	1	Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_makarova	0
+We had plans to include a quantitative comparison, however we were not able obtain a good dataset for doing so. It would be interesting to replicate Shepard’s experiments taking care to collect enough good quality images/movies for a quantitative analysis on the patterns.	2	1	The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_makarova	0
+Replicating Shepard and Sumner’s results was our primary objective for this paper, as we state in the discussion. In future work we would like to increase the scale of the simulations by adding more trichomes and a deeper domain to (hopefully) get more robust reticulate formation. We would also like to do a quantitative analysis on experimental data to in order to fit the model to the data. Finally, we would like to see whether this same minimal system plus photomovement is sufficient to produce the cone-shaped structures documented by Walter, Petroff and others. If successful, we could then link macroscopic features of similar stromatolites back to the parameters of the trichomes, as well as understand the contribution of each factor in the model to building these structures. We have added this explanation to the conclusions section.	2	1	Round 2: Author Response to Reviewer 1 We have addressed the reviwer’s requests: (1)			life4030433_makarova	0
+We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low. The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display. In the paper we justify (at some length) the bending modulus we used, which we extrapolated from various sources, and explain why we believe the value in Boal and Ng’s study is likely too low. The reviewer did not question the logic or the sources used to arrive at this value, but nevertheless seemed surprised at the result. The reviewer claims that the parameter falls outside of the physical regime, but doesn’t offer a justification other than that the implied persistence length “seems too large”. However, the large persistence length simply implies that the cyanobacteria are stiff enough that they are largely unaffected by thermal fluctuations. Despite this, the trichomes in our simulations are still quite flexible in practice, as can be seen in Figure 4. Even if the virtual trichomes are somewhat too stiff, we do not feel that this would have a major impact on the results.	2	1	However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			life4030433_perova	0
+We added the following statement to the end of Section 3.3: “The simulation results of the previous section for β > 0.5 are unlikely to be affected by the domain size, however, since the features in those simulations are on a much smaller scale than the domain size and are also more chaotic, as seen in the correlation length and the global alignment (Figure 5a,d).”	2	1	Thus, the length of a filament is not a model parameter.			life4030433_perova	0
+We added the following clarification to the end of the non-dimensionalization section: “In the subsequent sections, simulations for β = 0 use a modified interaction force such that there is no cohesion, but the hard core repulsion is maintained, i.e., 0 0 c F = for 0 h ≥ , otherwise 0 c c F F = with β = 0.125.”	2	1	Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			life4030433_perova	0
+We added two references to justify using the Lennard–Jones potential.	2	1	While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			life4030433_perova	0
+We added the following clarification: “This is simulated by generating a pseudo-random number x following a uniform distribution X ~ U(0,1), and reversing the gliding direction if x < ω⋅∆t.	2	1	They also explore behaviour in much of the available parameter space.			life4030433_perova	0
+We amended the following sentences of the discussion: “These differences may be due to the fact that we use a shallow domain (7.5 microns) to reduce the simulation run time, which may cause the pattern to be “squashed” as the domain ceiling prevents ridges from growing vertically” “Restricting the virtual trichomes such that they may only glide when in contact with the substratum or another trichome may promote further aggregation of the trichomes from streams into ridges, as a trichome would be less likely to successfully break away from a stream because it would lose much of its propulsive force as it lost contact with neighboring trichomes and/or the substratum.”	2	1	Thus, the length of a filament is not a model parameter.			life4030433_perova	0
+We added the following sentence to Section 2.4: “Gliding requires contact with some (semi-)solid substrate in order to provide a reaction force to the gliding mechanism. We assume that the trichomes are immersed in highly viscous medium that allows them to glide freely in all directions. This medium could consist of the EPS the trichomes produce copiusly when gliding [Hoiczyk2000].” (8)	2	1	Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			life4030433_perova	0
+The exact force is now given.	2	1	Rather, length is determined dynamically as ~ N k τ + (10)			life4030433_perova	0
+The bibliography now has volume and page numbers.	2	1	Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_perova	0
+The reviewer did not provide any examples where he/she felt a reference was warranted. We cited 55 sources, which we feel offers reasonable background and justification for our assumptions.	2	1	As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+).			life4030433_perova	0
+References to the movies were added to the results section.	2	1	Notably, the introduction of reproduction does not introduce another parameter.			life4030433_perova	0
+We added the following sentence to the end of Section 2.8: “This value corresponds to a persistence length of 0.49 m, which implies that the trichomes would be practically unaffected by thermal fluctuations.”	4	1	As a result, I do not recommend that this paper should be published in its current form.			life4030433_perova	0
+We added the following paragraph to the discussion: “Many of the model parameters, such as trichome length, diameter, gliding speed, reversal frequency, etc. are easily measured and are well known. However, the bending modulus is more difficult to measure directly. We attempted to use the bending modulus implied from the relation α= kBT ε and Boal and Ng’s (2010, p. 4625) measurements of the trichome persistence length, but the resulting value seemed too low compared to measurements of other bacteria and in practice the virtual trichomes appeared flaccid during simulations. It is possible that the flexure seen in Boal and Ng’s trichomes was due more to the motility of the trichomes than random thermal fluctuations, in which case the above relation would no longer be valid, and a more complex model would be required to associate the observed geometry of the trichomes to their bending modulus. For example, Wolgemuth (2005) used an elastic model to estimate the bending modulus of M. xanthus by fitting the model to the flailing motions of a Myxobacterium stuck at one end.”	4	1	Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			life4030433_perova	0
+The symbol used to denote trichome length was changed to an uppercase lambda. The , ⋅⋅ is often used to denote a tuple in computer science, in this case a pair. The superscripts should have been subscripts. The notation used in Equations (5)–(8) was adopted from Bergou et al’s paper. h-hat is in fact a unit vector parallel to h, and we’ve clarified the text to this point.	2	1	They also explore behaviour in much of the available parameter space.			life4030433_perova	0
+We added a few sentences to the introduction that expand a bit on similar structures in the Petroff and Walter references.	2	1	The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_perova	0
+Actually, as Reviewer 1 noted, we use a viscosity that is 1000 times greater than that of water (following Wolgemuth et al. 2005), for the very reasons the reviewer mentioned.	2	1	However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			life4030433_perova	0
+The statement has been clarified and is now: “The sum of the interaction’s opposing forces and torques is zero, thereby ensuring Newton's third law is respected.”	2	1	In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			life4030433_perova	0
+Our estimate is simplistic, however we were unable to find a reliable measurement of this parameter in the literature, and so we resorted to some simple extrapolations.	2	1	I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_perova	0
+There are indeed additional parameters. We have added a complete list to Section 2.8. Many are explored to some extent in the results (beta, reversal frequency, domain dimensions, trichome R7 density), whereas most of the physical characteristics of the trichomes are known from published sources (diameter, length, gliding speed, bending modulus), and finally N was simply chosen so that trichomes had a “smooth” appearance.	2	1	However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			life4030433_perova	0
+To avoid confusion on this point we have replaced this statement with the following: “We find that these errors are infrequent and acceptable as a trade-off for increased computational performance.” We have also added a comment to Section 2.5 regarding the Lennard–Jones interaction: “The attractive force has a short range, and is practically zero at h = 2θ.”	2	1	I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_perova	0
+It would be interesting to explore the approach the reviewer suggested; we have not yet explored more coarse grained models.	2	1	The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_perova	0
+Shepard et al. claim that the formation of the reticulate pattern occurs on a faster scale than cell growth and division, and so we did not consider this in our model. Including this might make the results more robust, however, and we added a comment to the discussion.	2	1	Qualitative comparisons are made to the fossil record and modern experiments.			life4030433_perova	0
+We had plans to include a quantitative comparison, however we were not able obtain a good dataset for doing so. It would be interesting to replicate Shepard’s experiments taking care to collect enough good quality images/movies for a quantitative analysis on the patterns.	2	1	As a result, I do not recommend that this paper should be published in its current form.			life4030433_perova	0
+Replicating Shepard and Sumner’s results was our primary objective for this paper, as we state in the discussion. In future work we would like to increase the scale of the simulations by adding more trichomes and a deeper domain to (hopefully) get more robust reticulate formation. We would also like to do a quantitative analysis on experimental data to in order to fit the model to the data. Finally, we would like to see whether this same minimal system plus photomovement is sufficient to produce the cone-shaped structures documented by Walter, Petroff and others. If successful, we could then link macroscopic features of similar stromatolites back to the parameters of the trichomes, as well as understand the contribution of each factor in the model to building these structures. We have added this explanation to the conclusions section.	2	1	I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_perova	0
+We are not sure if type IV pili are visible in our EM figures. They are very thin (less than 1 nm) and normally visualized by negative staining. In the published EM figures of Phormidium (conventional and cryo EM), type IV pili have not been noted. The filaments that we see in our EM figures could be components of sheath materials.	2	1	The text was revised according to the comments.			life4040819_makarova	0
+This was corrected.	2	1	A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			life4040819_makarova	0
+We did not find such an indication in the literature. This point was added in the text.	2	1	The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			life4040819_makarova	0
+The expression was modified.	2	1	Response: We thank the reviewers for giving us helpful comments.			life4040819_makarova	0
+Line 45—This was corrected.	2	1	The observations and models are consistent with previously published work on motility of various cyanobacteria and provide new insights into the mechanics of motility of cyanobacteria that are interesting.			life4040819_makarova	0
+Lines 268 and 269—These were corrected. We found many similar cases, too.	2	1	presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			life4040819_makarova	0
+Line 287—Two values were added.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_makarova	0
+Line 290—Some description was added.	2	1	ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_makarova	0
+Line 415—This was corrected.	2	1	Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			life4040819_makarova	0
+The choice of EM technique might have been wrong in visualizing the so-called oscillin fibrils, but was effective in visualizing membrane structures. The absence of “oscillin” was based on the genomic analysis but not on EM images. In fact, “oscillin” is a glycine-rich large protein which is poorly conserved in bacteria. In the reported case of P. uncinatum, oscillin might be important in forming fibrils, but in other organisms, other proteins could function as surface fibrils aligning the flow of slime. We agree that the method of fixation was not good for preserving surface structures such as oscillin fibrils, if present.	2	1	Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			life4040819_makarova	0
+This point was added in Section 3.7, in which oscillin is discussed. In general, pore-inclination and oscillin fibrils are not exclusive.	2	1	The contribution of motility to the formation of supracellular structure is generally an area of research regarding cyanobacteria that has been overlooked.			life4040819_makarova	0
+The quality of Figure 1A was rather bad. The contrast was corrected and a new figure was inserted. In the new Figure 1A, the sheath is quite visible. The quality of EM figure is very low as embedded in an MS word file. We provide a better quality figure as supplemental Figure S1. The use of “sheath” is therefore correct in the paragraph describing the figure. In the model, the word “sliding” was not understood in the sense in which we wanted it to be. This is lateral sliding that provokes curvature of the filament. This point was clarified in the text.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_makarova	0
+What is “the basal body part”? Basal body is an organelle of eukaryotic cell. If this means the connection of junctional pore tube and inner membrane, it is clearly seen in the figure, as well as in old paper by Halfen and Castenholz. A high quality figure is provided as Supplementary Figure S1. The “bulges” in the inner membrane are also visible. In the Line 183, the description was revised.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_makarova	0
+In Figure 3D, the junctional pores are seen as an array of tilted tubes. The mechanism of rotation is still a mystery because oscillin itself is not present in the strain KS. This is discussed in Section 3.7 with reservations on our hypothesis of inclined junctional pores. On the other hand, the involvement of fibrils in the rotation and locomotion was proposed a long time ago, whether the fibrils are made of oscillin or other proteins. In this respect, we still have to work a lot to identify the real mechanism of rotation.	2	1	Response: We thank the reviewers for giving us helpful comments.			life4040819_makarova	0
+This is not a good way of naming headings. A heading should not be a phrase or conclusion, but should be descriptive words.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_makarova	0
+This was described in Section 3.7.	2	1	Round 1: and Author Response The manuscript “Cellular dynamics drives emergence of supracellular structure in the cyanobacterium Phormidium sp.			life4040819_makarova	0
+This point was corrected.	2	1	presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			life4040819_makarova	0
+There might be a misunderstanding in the first part. The rotation of filament does not drive macroscopic rotation or spiral formation. What we describe in the text was the switch to turn to the left was governed by the filament rotation, but the real formation of a spiral is driven by the locomotion of the filament. The cited paper described the clumping of Anabaena cylindria in a dense culture. The clumping or aggregation in Arthrospira (Ohmori group) is mediated by cAMP, but A. cylindrica might be different. I have been using Anabaena for about 40 years, but I have never seen Anabaena filaments form a spiral on agar plates. Spiral formation and clumping are different phenomena. It is difficult to discuss the relationship (if any) between clumping and rotation.	2	1	Round 1: and Author Response The manuscript “Cellular dynamics drives emergence of supracellular structure in the cyanobacterium Phormidium sp.			life4040819_makarova	0
+The current paper is focused on Phormidium. We never used Myxobacteria, and we have no idea about the motility in Myxobacteria. To clarify the situation, the mention to Myxobacteria was added in the text.	2	1	A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			life4040819_makarova	0
+Lines 94–95: We have determined genomic sequences of many organisms, but it is not easy to publish the genomes as genome paper. The sequences should be connected by PCR, and annotated. In the current study, we are interested in the genes involved in motility. We annotated the related genes but we would not connect the contigs and annotate all the genes. Nowadays, every researcher can sequence his/her own materials quite easily. It is not necessary to deposit all the raw sequence data.	2	1	In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			life4040819_makarova	0
+Lines 227–230: This was corrected.	2	1	ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_makarova	0
+Lines 316–317: We did not notice this point. Polysaccharide chain may not be very long. Many of the products of the hps gene cluster encode glycosyltransferases and pseudopilins, which are, respectively, involved in the synthesis and secretion of the slime. The secretion of slime is likely mediated by some molecular machinery that is otherwise involved in type II secretion/motility machinery. Reference 24 was added.	2	1	Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			life4040819_makarova	0
+We thank the reviewers for giving us helpful comments. This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript. The text was revised according to the comments. In addition, Figure 3A was replaced by a figure of better contrast. Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement. Within the text, corrected words are highlighted.	2	1	ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_perova	0
+We are not sure if type IV pili are visible in our EM figures. They are very thin (less than 1 nm) and normally visualized by negative staining. In the published EM figures of Phormidium (conventional and cryo EM), type IV pili have not been noted. The filaments that we see in our EM figures could be components of sheath materials.	2	1	In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			life4040819_perova	0
+This was corrected.	2	1	(3) In the discussion there are further points that need to be addressed: (a)			life4040819_perova	0
+We did not find such an indication in the literature. This point was added in the text.	2	1	Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			life4040819_perova	0
+The expression was modified.	2	1	Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			life4040819_perova	0
+Line 45—This was corrected.	2	1	A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			life4040819_perova	0
+ Lines 268 and 269—These were corrected.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_perova	0
+ Line 287—Two values were added.	2	1	Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			life4040819_perova	0
+ Line 290—Some description was added.	2	1	R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			life4040819_perova	0
+ Line 415—This was corrected.	2	1	Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			life4040819_perova	0
+The choice of EM technique might have been wrong in visualizing the so-called oscillin fibrils, but was effective in visualizing membrane structures. The absence of “oscillin” was based on the genomic analysis but not on EM images. In fact, “oscillin” is a glycine-rich large protein which is poorly conserved in bacteria. In the reported case of P. uncinatum, oscillin might be important in forming fibrils, but in other organisms, other proteins could function as surface fibrils aligning the flow of slime. We agree that the method of fixation was not good for preserving surface structures such as oscillin fibrils, if present.	2	1	However, a number of shortcomings need to be addressed before the manuscript could be accepted for publication.			life4040819_perova	0
+This point was added in Section 3.7, in which oscillin is discussed. In general, pore-inclination and oscillin fibrils are not exclusive.	2	1	The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			life4040819_perova	0
+There is no clear distinction in the text between the “sheath” and the secreted “slime” of the cells, which are two completely different structures both physically and chemically (Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Figure 1A shows a filament that lacks the “sheath”, while in other pictures the structure is visible. The reason for this is the following: cyanobacteria of the genus Phormidium build over time a carbohydrate layer on their surface that is physically attached to their cell surface, usually called the sheath. Filaments that are ensheathed are non-motile! In contrast, the slime that is secreted by gliding filaments of the same species is not visible in TEM preparations, even when cryo-preservation methods are used (see Figure 1A of the ms and Envelope structure of four gliding filamentous cyanobacteria. J. Bacteriol. 1995, 177, 2387–2395 and Structural and biochemical analysis of the sheath of Phormidium uncinatum. J. Bacteriol. 1998, 180, 3923– 3932). Therefore, the authors should go through their ms and make a careful distinction between these two structures. For example, in the discussion it sounds as if the “sheath” is preventing the R5 cells from sliding, however this is the slime tube that is secreted that is not physically attached to the cell surface at all.	2	1	The contribution of motility to the formation of supracellular structure is generally an area of research regarding cyanobacteria that has been overlooked.			life4040819_perova	0
+What is “the basal body part”? Basal body is an organelle of eukaryotic cell. If this means the connection of junctional pore tube and inner membrane, it is clearly seen in the figure, as well as in old paper by Halfen and Castenholz. A high quality figure is provided as Supplementary Figure S1. The “bulges” in the inner membrane are also visible. In the Line 183, the description was revised.	2	1	Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			life4040819_perova	0
+In Figure 3D, the junctional pores are seen as an array of tilted tubes. The mechanism of rotation is still a mystery because oscillin itself is not present in the strain KS. This is discussed in Section 3.7 with reservations on our hypothesis of inclined junctional pores. On the other hand, the involvement of fibrils in the rotation and locomotion was proposed a long time ago, whether the fibrils are made of oscillin or other proteins. In this respect, we still have to work a lot to identify the real mechanism of rotation.	2	1	By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			life4040819_perova	0
+This is not a good way of naming headings. A heading should not be a phrase or conclusion, but should be descriptive words.	2	1	A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			life4040819_perova	0
+This was described in Section 3.7.	2	1	The contribution of motility to the formation of supracellular structure is generally an area of research regarding cyanobacteria that has been overlooked.			life4040819_perova	0
+This point was corrected.	2	1	presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			life4040819_perova	0
+There might be a misunderstanding in the first part. The rotation of filament does not drive macroscopic rotation or spiral formation. What we describe in the text was the switch to turn to the left was governed by the filament rotation, but the real formation of a spiral is driven by the locomotion of the filament. The cited paper described the clumping of Anabaena cylindria in a dense culture. The clumping or aggregation in Arthrospira (Ohmori group) is mediated by cAMP, but A. cylindrica might be different. I have been using Anabaena for about 40 years, but I have never seen Anabaena filaments form a spiral on agar plates. Spiral formation and clumping are different phenomena. It is difficult to discuss the relationship (if any) between clumping and rotation.	2	1	The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			life4040819_perova	0
+The current paper is focused on Phormidium. We never used Myxobacteria, and we have no idea about the motility in Myxobacteria. To clarify the situation, the mention to Myxobacteria was added in the text.	2	1	Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			life4040819_perova	0
+Lines 94–95: We have determined genomic sequences of many organisms, but it is not easy to publish the genomes as genome paper. The sequences should be connected by PCR, and annotated. In the current study, we are interested in the genes involved in motility. We annotated the related genes but we would not connect the contigs and annotate all the genes. Nowadays, every researcher can sequence his/her own materials quite easily. It is not necessary to deposit all the raw sequence data.	2	1	Within the text, corrected words are highlighted.			life4040819_perova	0
+Lines 227–230: This was corrected.	2	1	The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			life4040819_perova	0
+Lines 316–317: We did not notice this point. Polysaccharide chain may not be very long. Many of the products of the hps gene cluster encode glycosyltransferases and pseudopilins, which are, respectively, involved in the synthesis and secretion of the slime. The secretion of slime is likely mediated by some molecular machinery that is otherwise involved in type II secretion/motility machinery. Reference 24 was added.	2	1	A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			life4040819_perova	0
+The three references have been noticed with gratitude and included in the MS.	2	1	In light of recent research developments and argumentation (some of it reviewed), his views should be considered a welcome addition to the many ideas that populate the “origin of life” field of inquiry that counter the dominant paradigm.			life4041050_makarova	0
+The 1998 paper leaves much to be desired. Its deficiencies reflect the excitement of the first hour. The state of the art at that time may be gleaned from an authoritative paper that came to the opposite conclusion: A.R. Mushegian and E.V. Koonin “Gene order is not conserved in bacterial evolution”. TIG 1996, 12, 289–290. The gene cluster table of 1998 was mainly retrieved from the annotations in published genomes and constructed manually with paper and pencil. The state of information technology at that time is reflcted by the fact that the table was folded an individually pasted by hand into each issue by the publisher.	2	1	Round 1: and Author Response The manuscript comprises an exciting account on the origin of life with emphasis on the emergence and function of RNA.			life4041050_makarova	0
+The 1998 paper contemplated speculatively a combination of small-scale gene doubling (as evidenced by the immediate neighborhood of EF-Tu/EF-G) and of a large-scale gene cluster doubling (as evidenced by the spacing between secE/secY and rpoH-A/rpoD) with the hope of a future deeper understanding based on folding structures. Now the referee makes an exciting suggestion that may, if executed successfully, go some way to satisfy that hope.	2	1	I have only a few comments, which might improve the manuscript.			life4041050_makarova	0
+The self-cleavage of RNA by 2'-OH is a chemical textbook fact. The 2'-OH group has the proper position and orientation for a nucleophilic attack on the phosphate bridge. The kinetics of the reaction is greatly favored by the 5-membered ring structure of the resulting cyclic phospho-bisester. The length of the RNA molecule is not relevant since each nucleophilic attack causes destruction of the chain. The effect should not be confused with the length-dependent “error catastrophe” of accumulating mutations of RNA. Incidentally, some anaerobic ribonucleotide reductases are ancient, while others (aerobic ones) are later inventions.	2	1	I have only a few comments, which might improve the manuscript.			life4041050_makarova	0
+Genome rearrangements are certainly important during later evolution of the phyla. At the level of LUCA, i.e., prior to the splitting of the domains, it is not clear, if and to what extent rearrangements of the modern style occurred. In this regard we should bear in mind that the LUCA genome may have exhibited sense-antisense coding on both strands as suggested by Rodin and Carter. This position has been adopted and discussed in the present paper. Therefore, speculations concerning possible genome rearrangements may be a bridge too far.	2	1	He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			life4041050_makarova	0
+The present analysis comes to the conclusion that the sets of canonical amino acids and bases as well as the genetic triplet code were largely complete at the level of LUCA. The Wong coevolution theory of the genetic code has been discussed in detail. The literature comprises numerous proposals concerning the origin of translation and other aspects of the genetic machinery. A review of all these proposals and many others is beyond the scope of the present paper. The present paper is a research paper and not a review paper. It aims at a comprehensive account of early evolution from the origin of life all the way to LUCA. This puts a systematic constraint on literature selection. An effort (unfortunately fallible) has been made to include all those references that integrate with the main lines of the present account into a coherent account. Contributions by others that have the character of theoretical modules that fit well into this account have been termed “theorems” with names of the main authors attached. A reference to a paper on nucleotide biosynthesis phylogeny is now cited in Section 7.	2	1	Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			life4041050_makarova	0
+Based on the valuable criticism Section 2 has been extensively revised. Terminology has been clarified. This Section has a rather restricted purpose. It provides chemical arguments for the proposition that the pioneer organisms could only exist at high temperature and that the subsequent forms of life remained hyperthermophilic for a long time until much later an irreversible evolution generated organisms that required lower and lower temperatures. This conclusion places severe constraints on all aspects of the evolution of the genetic machinery. The fascinating topics of thermodynamics, energy dissipation and information are outside the scope of the paper.	2	1	The alternative views he previously generated are much expanded in this manuscript.			life4041050_makarova	0
+The author expresses his gratitude for the two additional references, which have been included in the text.	2	1	He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			life4041050_makarova	0
+The statement has been clarified.	2	1	Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			life4041050_makarova	0
+The author prefers the present title, because the paper is concerned with the origin of the genetic machinery. Theories on the origin of life or on other aspects of early evolution, such as cellularization, serve merely as starting points.  In the introduction the term “or proteins” has been added after “RNA” ,  and the term “retrodict” has been defined.  The term “interpolate” has been clarified.  The problem of mutational saturation is now included in the discussion of Figure 1.  The term “multiply impaired” has been replaced by a clearer wording.  LUCA is discussed only in Section 1.  The protein cycle is discussed in a separate Section 4, which is concerned with the course of evolution before LUCA.	2	1	I must note that a careful evaluation of all facets requires expertise in a multitude of disciplines (from prebiotic chemistry and structural biology to evolutionary bioinformatics and biochemistry) and considerable time, none of which I possess.			life4041050_makarova	0
+There are many relevant literature references. Among these there are specific proposals concerning a circumscribed problem that are included to fill a logical gap, as a theoretical module so to speak. These have been designated by the term “theorem” with the added name(s) of the main R7 author(s). This makes it clear that the account given is comprehensive in the sense that major independent contributions by other scientists integrate readily with the overall account given.	2	1	I must note that a careful evaluation of all facets requires expertise in a multitude of disciplines (from prebiotic chemistry and structural biology to evolutionary bioinformatics and biochemistry) and considerable time, none of which I possess.			life4041050_makarova	0
+The relationship of His and Trp biosyntheses has been toned down.	2	1	Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			life4041050_makarova	0
+Section 8 has been revised in order to address the issues involved in the last query.	2	1	I must note that a careful evaluation of all facets requires expertise in a multitude of disciplines (from prebiotic chemistry and structural biology to evolutionary bioinformatics and biochemistry) and considerable time, none of which I possess.			life4041050_makarova	0
+The concern the reviewer mentions here was also one of our main conclusions for the article. Apparently we did not formulate it well enough. It has thus been reformulated to: “However an accurate taxonomy can never be achieved by the use of a barcoding method only, since it is based on nucleotide substitutions of a single gene. Accurate classification always requires an integrative taxonomy effort R2 including characteristics from ecology, morphology and physiology, as already previously suggested for prokaryotes [9,28,29,45], and as it is becoming common also for animal taxonomy (e.g., [53,54]).” Moreover, it is necessary to mention the following aspects of this manuscript: (1)	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+We understand that our choice of naming as in the database needed clarification. We have thus added the following paragraph. We thank the reviewer for the useful example and have included it: “The names of genera and species used in this study where copied form the names given in the database. This choice was made for the reason that cyanobacterial phylogeny underlies continuous changes and genera and species are often reclassified (e.g., [34]). Thus a whole different type of work would be required in order to use all state-of-art classification of cyanobacteria. Moreover, for the aims of this study not so much the phylogenetic classification of the bacterial species, but the genetic structure of the cyanobacterial 16S rRNA genes was of importance. The use of the names provided in the database on the other hand enables other researchers to conduct a similar analysis using the same sequences. Thus some old and revised names are used, for example the species name Anabaena bergii is used throughout the study despite its revised taxonomy in the genus Crysosporum [35].” (2)	2	1	We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+We understand that particularly the case of Synechoccocus needs clarification and have thus extended the explanation there: “Sequences termed Synechococcus on the other hand were analysed together with sequences of the monophyletic clade Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included, too”.	2	1	Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_makarova	0
+We agree with the reviewer that we have to avoid misunderstandings of this kind and added a very clear statement to the discussion: “Moreover it has to be emphasised that a close analysis of the actual properties of the cyanobacterial groups tested is limited by the fact that we used the provided R3 names for the sequences and groups. Thus this study intends only to verify the existence of a barcoding gaps within the 16S rRNA sequences of certain cyanobacterial groups, and by no means revise or confirm the complex cyanobacterial taxonomy.”	2	1	Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+We agree with the reviewer that we have not stressed this fact enough and have added more sentences explaining why OTUs are used and the problem: “Enormous progress happened in prokaryote taxonomy in the recent years [8,12,24]. However, the data produced with novel methodologies, such as next generation sequencing, often requires a high throughput taxonomic classification of sequences such as fixed threshold to identify OTUs. This kind of fixed numeric classifications can always only be a vague approximation to the actual structure of relatedness of organisms.”	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gap existed at all.	2	1	We thus only briefly discuss names and taxonomy in our analysis.			life5010050_makarova	0
+we now include new citations in relationship to different problems: Butlin et al. (2009), Cohan (2011), Cohan (2013), Cohan & Aracena (2012), Diekmann et al. (2004), Nosil (2012), Vos (2011), Wiedenbeck & Cohan (2011).	2	1	Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+We agree with the reviewer’s concern. In fact, this is exactly the reason why we chose Cyanobacteria for this analysis. We reformulated this part of the introduction, to make this statement more clear, which now reads: “We choose Cyanobacteria as an example of prokaryotes, not because they are representative for all prokaryotes, but because there is ample phenotypic, ecological, physiological ultrastructural, and biochemical evidence of the existence of independently evolving units in this group [20,21]. Thus, the expectation is that, if a barcoding gap exists in prokaryotes, this should be more easily seen in taxa where groups can be identified also with other methods, as in Cyanobacteria.”	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+We apologize for our carelessness in double-checking the naming of the organisms and hope that we now eliminated all errors.	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+We agree with the reviewer, this part was formulated clumsily. What we meant was actually exactly what the reviewer means. The two groups were polyphyletic and we could only chose a monophyletic group when analysing them together. We changed the sentence to: “… by two genera that were monophyletic only when taken together in the database used, e.g., Leptolyngbia and Chamaesiphon …” In accordance we did not check for relatedness of the taxa, neither in this one nor for any other one.	2	1	We thus only briefly discuss names and taxonomy in our analysis.			life5010050_makarova	0
+We agree that this might be confusing to the reader. We have therefore clarified this choice and added this part to the first M&M section and the literature suggested has been cited: “Sequences termed Synechococcus on the other hand were analysed together with sequences of Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included too.”	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+The sequences we used were clustering monophyletically a-priori in the tree provided in the database used, and we chose them independently of the name. We now clarified this in the first section of M&M: “Thereby a group was considered monophyletic if it was monophyletic in the tree provided with the database, regardless of the taxonomy and the nomenclature of the organisms included in the clade (Supplemetary Figure 1), and only secondarily if there was a correspondence with known taxa.”	2	1	ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			life5010050_makarova	0
+As mentioned above, we did not only choose sequences that had the same name, but groups of sequences that were monophyletic in the database tree. Thus we should not have artificially introduced a barcoding gap by sequence selection.	2	1	To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			life5010050_makarova	0
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names. We thus only briefly discuss names and taxonomy in our analysis. Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point. We hope that now both reviewers, and all other potential readers, will not be misled by our approach. We do not deal with taxonomy at all, only with testing the presence of a barcoding gap. Similarly to the replies for the previous reviewer, our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gaps existed at all. Given this rationale and the kind of data we used, no clear statement towards the two barcoding groups within Cylindrospermopsis can be made. Nearly all sequences within the database are termed Cylindrospermopsis raciborskii by the people who deposited the sequences. However the clade does not contain the type-strain (which was not in the database). So we would rather not speculate too much on the taxonomic naming of these and other barcoded groups. However, we can clearly say that what is deposited in database SILVA 111 as Cylindrospermopsis contains two species with a barcoding gap. For Planktothrix we added a sentence: “Considering the naming of Planktothrix sequences in the database, some OTUs and ABGD units seem to correspond not only to monophyletic groups but also to named species such as P. mougeotii or similarly in the case of Fischerella muscicola (Figure 3).” R6 However we wish to remain careful on these kinds of statements since we are not sure if the underlying naming of the species used is correct or not.	2	1	Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+We understand why the reviewer would think that that would be beneficial. However, one has to keep in mind that we were not constructing a phylogenetic tree for all cyanobacteria but for single genera. In this case it is preferential to use a sister group that is more closely related to gain resolution within that group. e.g., Hedtke, S.M. ; Townsend, T.M. ; Hillis, D.M. Resolution of phylogenetic conflict in large data sets by increased taxon sampling. Syst. Biol. 2006, 55, 522–529. Moreover, for the sake of the barcoding analyses, the choice of the outgroup will not affect the results, given that it will not change the relationships towards the tips of the tree.	2	1	We thus only briefly discuss names and taxonomy in our analysis.			life5010050_makarova	0
+The last paragraph of the discussion already included some comments on this issue. Now the paragraph has been expanded to provide a more detailed discussion on the possibility of using a DNA barcoding gap in cyanobacteria.	2	1	We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+We apologize for our carelessness in double checking the naming of the organisms and hope that we now eliminated all errors.	2	1	We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+The concern the reviewer mentions here was also one of our main conclusions for the article. Apparently we did not formulate it well enough. It has thus been reformulated to: “However an accurate taxonomy can never be achieved by the use of a barcoding method only, since it is based on nucleotide substitutions of a single gene. Accurate classification always requires an integrative taxonomy effort R2 including characteristics from ecology, morphology and physiology, as already previously suggested for prokaryotes [9,28,29,45], and as it is becoming common also for animal taxonomy (e.g., [53,54]).”	2	1	We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_perova	0
+We understand that our choice of naming as in the database needed clarification. We have thus added the following paragraph. We thank the reviewer for the useful example and have included it: “The names of genera and species used in this study where copied form the names given in the database. This choice was made for the reason that cyanobacterial phylogeny underlies continuous changes and genera and species are often reclassified (e.g., [34]). Thus a whole different type of work would be required in order to use all state-of-art classification of cyanobacteria. Moreover, for the aims of this study not so much the phylogenetic classification of the bacterial species, but the genetic structure of the cyanobacterial 16S rRNA genes was of importance. The use of the names provided in the database on the other hand enables other researchers to conduct a similar analysis using the same sequences. Thus some old and revised names are used, for example the species name Anabaena bergii is used throughout the study despite its revised taxonomy in the genus Crysosporum [35].” (2)	2	1	Round 2: I accept the paper in the present form.			life5010050_perova	0
+We understand that particularly the case of Synechoccocus needs clarification and have thus extended the explanation there: “Sequences termed Synechococcus on the other hand were analysed together with sequences of the monophyletic clade Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included, too”.	2	1	Thus this study intends only to verify the existence of a barcoding gaps within the 16S rRNA sequences of certain cyanobacterial groups, and by no means revise or confirm the complex cyanobacterial taxonomy.”			life5010050_perova	0
+We agree with the reviewer that we have to avoid misunderstandings of this kind and added a very clear statement to the discussion: “Moreover it has to be emphasised that a close analysis of the actual properties of the cyanobacterial groups tested is limited by the fact that we used the provided R3 names for the sequences and groups.	2	1	I believe that the article with proposed changes will be useful for future research.			life5010050_perova	0
+We agree with the reviewer that we have not stressed this fact enough and have added more sentences explaining why OTUs are used and the problem: “Enormous progress happened in prokaryote taxonomy in the recent years [8,12,24]. However, the data produced with novel methodologies, such as next generation sequencing, often requires a high throughput taxonomic classification of sequences such as fixed threshold to identify OTUs. This kind of fixed numeric classifications can always only be a vague approximation to the actual structure of relatedness of organisms.”	2	1	Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_perova	0
+our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gap existed at all.	2	1	Of course, the complex of all these methods must be corrected with the progress of science, and especially for cyanobacteria is necessary to apply the so called “polyphasic approach” for their classification.			life5010050_perova	0
+we now include new citations in relationship to different problems: Butlin et al. (2009), Cohan (2011), Cohan (2013), Cohan & Aracena (2012), Diekmann et al. (2004), Nosil (2012), Vos (2011), Wiedenbeck & Cohan (2011).	2	1	We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_perova	0
+We agree with the reviewer’s concern. In fact, this is exactly the reason why we chose Cyanobacteria for this analysis. We reformulated this part of the introduction, to make this statement more clear, which now reads: “We choose Cyanobacteria as an example of prokaryotes, not because they are representative for all prokaryotes, but because there is ample phenotypic, ecological, physiological ultrastructural, and biochemical evidence of the existence of independently evolving units in this group [20,21]. Thus, the expectation is that, if a barcoding gap exists in prokaryotes, this should be more easily seen in taxa where groups can be identified also with other methods, as in Cyanobacteria.”	2	1	ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			life5010050_perova	0
+We apologize for our carelessness in double-checking the naming of the organisms and hope that we now eliminated all errors.	2	1	Round 2: I accept the paper in the present form.			life5010050_perova	0
+We agree with the reviewer, this part was formulated clumsily. What we meant was actually exactly what the reviewer means. The two groups were polyphyletic and we could only chose a monophyletic group when analysing them together. We changed the sentence to: “… by two genera that were monophyletic only when taken together in the database used, e.g., Leptolyngbia and Chamaesiphon …” In accordance we did not check for relatedness of the taxa, neither in this one nor for any other one.	2	1	I believe that the article with proposed changes will be useful for future research.			life5010050_perova	0
+We agree that this might be confusing to the reader. We have therefore clarified this choice and added this part to the first M&M section and the literature suggested has been cited: “Sequences termed Synechococcus on the other hand were analysed together with sequences of Prochlorococcus, since Synechococcus is known to be a polyphyletic group and if all sequences (including a minimum of five lineages [36–38]) are taken together, Prochlorococcus has to be included too.”	2	1	Of course, the complex of all these methods must be corrected with the progress of science, and especially for cyanobacteria is necessary to apply the so called “polyphasic approach” for their classification.			life5010050_perova	0
+The sequences we used were clustering monophyletically a-priori in the tree provided in the database used, and we chose them independently of the name. We now clarified this in the first section of M&M: “Thereby a group was considered monophyletic if it was monophyletic in the tree provided with the database, regardless of the taxonomy and the nomenclature of the organisms included in the clade (Supplemetary Figure 1), and only secondarily if there was a correspondence with known taxa.”	2	1	Second Round of Evaluation Round 2: I have had the more critical remarks to the manuscript; however, I think that the presented data are useful and should be published.			life5010050_perova	0
+As mentioned above, we did not only choose sequences that had the same name, but groups of sequences that were monophyletic in the database tree. Thus we should not have artificially introduced a barcoding gap by sequence selection.	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_perova	0
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names. We thus only briefly discuss names and taxonomy in our analysis. Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point. We hope that now both reviewers, and all other potential readers, will not be misled by our approach. We do not deal with taxonomy at all, only with testing the presence of a barcoding gap. Similarly to the replies for the previous reviewer, our aim was not to start from known and named sequences and test whether species from DNA taxonomy matched them or not; our aim was to test whether a barcoding gaps existed at all. Given this rationale and the kind of data we used, no clear statement towards the two barcoding groups within Cylindrospermopsis can be made. Nearly all sequences within the database are termed Cylindrospermopsis raciborskii by the people who deposited the sequences. However the clade does not contain the type-strain (which was not in the database). So we would rather not speculate too much on the taxonomic naming of these and other barcoded groups. However, we can clearly say that what is deposited in database SILVA 111 as Cylindrospermopsis contains two species with a barcoding gap. For Planktothrix we added a sentence: “Considering the naming of Planktothrix sequences in the database, some OTUs and ABGD units seem to correspond not only to monophyletic groups but also to named species such as P. mougeotii or similarly in the case of Fischerella muscicola (Figure 3).” R6 However we wish to remain careful on these kinds of statements since we are not sure if the underlying naming of the species used is correct or not.	2	1	Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_perova	0
+We understand why the reviewer would think that that would be beneficial. However, one has to keep in mind that we were not constructing a phylogenetic tree for all cyanobacteria but for single genera. In this case it is preferential to use a sister group that is more closely related to gain resolution within that group. e.g., Hedtke, S.M. ; Townsend, T.M. ; Hillis, D.M. Resolution of phylogenetic conflict in large data sets by increased taxon sampling. Syst. Biol. 2006, 55, 522–529. Moreover, for the sake of the barcoding analyses, the choice of the outgroup will not affect the results, given that it will not change the relationships towards the tips of the tree.	2	1	Moreover, it is necessary to mention the following aspects of this manuscript: (1)			life5010050_perova	0
+The last paragraph of the discussion already included some comments on this issue. Now the paragraph has been expanded to provide a more detailed discussion on the possibility of using a DNA barcoding gap in cyanobacteria.	2	1	The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			life5010050_perova	0
+We apologize for our carelessness in double checking the naming of the organisms and hope that we now eliminated all errors.	2	1	Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_perova	0
+This review does not require any action on our part.	2	1	Round 1: and Author Response The article “Emergent Chemical Behaviour in Variable-Volume Protocells” focuses on the consequences that a changing volume may have on a set of reactions encapsulated within a semipermeable vesicle; in particular, this kind of compartment can degenerate existing bistable reactions, or promote emergent bistability from very simple reactions, which are not bistable in bulk conditions.			life5010181_perova	0
+Here, the reviewer did not like our use of the particular words “novelty” and “innovation”. o Use of the word novelty − Instances of “chemical novelty” in the manuscript replaced by “emergent chemical behaviour”. − On Page 8, after Equation 8, we removed sentence: The emergence of bistability will serve as a proxy for the emergence of other chemical novelties in the vesicle reactor model. Replaced whole paragraph, to read: In particular, in these initial stages, we will focus on the emergence of bistability in the vesicle reactor model—a dynamical feature deducible directly from the number and stability of the fixed points present (i.e., two asymptotically stable points separated by an unstable saddle point). We also expect that more complicated dynamical regimes could also be present in the model, like multi-stability or global phase space features such as limit cycles giving rise to sustained oscillations. However, investigation of these regimes will be deferred to later work: for the time being, the “emergent chemical behavior” referred to in the title will be restricted to bistability. We think this is a clearer explanation, and also uses the word “regime” suggested by the reviewer. o Use of the word innovation We kept the 2 occurrences of the word “innovation” in the abstract and the introduction. An innovation is defined as a “new method, idea or product” in the dictionary. We use the word to refer to new emergent chemical behaviour that the whole vesicle system exhibits (e.g., expanded steady states), which did not exist before. We think the use of this word is acceptable.	2	1	Remarkably, even reaction systems that have no chemical species in common could become R2 indirectly coupled to each other through the volume they share, by means of a sort of osmotic coupling.			life5010181_perova	0
+we added a new footnote, to explain our standing on this issue: Reference 36: In this work, concentrations outside the vesicle are set as system parameters. However, we make no commitment to the type of environment the vesicle is embedded in or how these concentrations are maintained. Our purpose is simply to show that bistability can exist in the model for certain sets of outside concentrations. Exploration of the model in explicit environments is deferred to future work.	2	1	Remarkably, even reaction systems that have no chemical species in common could become R2 indirectly coupled to each other through the volume they share, by means of a sort of osmotic coupling.			life5010181_perova	0
+Reference 11 has been added after Equation 10, to point the reader to where to find information about phi limits was first discussed. Repeating this information here would complicate the paper. The lines Phi = 2^(1/3) and 4^(1/3) in Figure 1d have now been explained.	2	1	Produces a dynamic model for chemical evolution rather than traditional deterministic, static, systems that requires centralised information to direct evolution, such as a constant gradient.			life5010181_perova	0
+It is difficult to make a direct and meaningful comparison like this. Without a container, the reaction system has less parameters, and thus the sampled parameter space is smaller. The aim in the paper, was to simply show that encapsulating a Schlogl model which was bistable in bulk condition, seemed to destroy this bistability.	2	1	As the authors acknowledge, the article is the first step toward a more complete understanding of an interesting class of possible chemo-physical phenomena.			life5010181_perova	0
+The minor issues were fixed.	2	1	Remarkably, even reaction systems that have no chemical species in common could become R2 indirectly coupled to each other through the volume they share, by means of a sort of osmotic coupling.			life5010181_perova	0
+This immunoquantitation method used is described in detail in the reference given (Brown et al. 2008). The reviewer is directed in particular to the Supplementary Methods section. To allow readers to get a better understanding of the method in this manuscript we have revised this part of the Experimental Section, adding extra text to increase clarity concerning the method.	2	1	The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+Since submitting the manuscript, we have accumulated a set of 84 parallel measurements of steady state oxygen evolution and the functional content of PSII measured using flash yields (with a solid state optode) and simultaneous FRR chlorophyll fluorescence induction curves, from which we can extract e- PSII-1 s-1, for both Synechococcus and Prochlorococcus cultures. Consistent with the long literature history of such measurements (ex. Suggett et al. 2004, 2009), we observe a good correlation between the two measures of electron transport per PSII. Plotting the FRR estimate of e- PSII-1 s-1 versus the O2 evolution/PSII content estimate of e- PSII-1 s-1 gives a slope of 1.26 and an R2 of 0.58. We have added this information to the Materials and Methods to support our use of FRR estimates of electron transport per PSII. Furthermore, our estimates of ETRmax from FRR induction curves are independently validated by the close correlation between ETRmax and 1/tau, shown in Figure 4A, since 1/tau is derived from the rate constant for the decay of fluorescence after induction, and thus does not depend (computationally) upon our estimator for ETRmax.	2	1	This is done for a number of globally significance marine microbes of Syn and Pro lineage.			life5010403_makarova	0
+This has been addressed with the addition of text in the abstract, introduction and discussion specifying that the cultures were grown under low light conditions.	2	1	The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+We have removed the word “nondiazotrophic” as we agree that it is not relevant to the discussion.	2	1	The data presented is robust and worthy of publication but should be discussed more critically with regard to published literature (outlined below) I suggest some minor suggestions that should be incorporated before publications.			life5010403_makarova	0
+We mean that they numerically dominate and we have clarified this in the introduction.	2	1	Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			life5010403_makarova	0
+We have clarified this statement in the text. We refer to the nitrogen cost in the form of allocation to protein per pigment bound.	2	1	Second Round of Evaluation Round 2: and Author Response			life5010403_makarova	0
+We are referring to the number of RUBISCO active sites measured by immunoquantitation. We are not referring to measured RUBISCO activity. We use this expression to be clear that we are referring to RbcL subunits rather than oligomeric RUBISCO. We have added a parenthetic phrase to make this more clear.	2	1	Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			life5010403_makarova	0
+We thank the reviewer for drawing this study to our attention. We have added a paragraph to the discussion to compare and contrast the Sukenik work with that presented here. The Sukenik data support a strong positive correlation between 1/tau and the RUBISCO to PSU ratio over a series of growth irradiances. While we also see a strong positive correlation between 1/tau and RUBISCO to PSII our results differ as the molar ratios of the components of the PSU differ significantly between the strains analyzed in the current work. This allows us to pinpoint the relationship of PSII to RUBISCO rather than other subunits of the PSU as the determinant of electron transport rate.	2	1	The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+We have added text to the end of the discussion to address this comment and the relevant reference has been added. Thank you for this suggestion.	2	1	provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			life5010403_makarova	0
+We have added a supplemental figure (Supplemental Figure #2) to show this calibrations curve. As all experiments presented here were performed under iron replete conditions, no iron starvation was performed.	4	1	provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			life5010403_makarova	0
+We have added a supplemental figure (Supplemental Figure #1) that shows the method applied with a sample blot, calibration curve and data analysis. This work required dozens of blots, each with its own standard curve, so it would not be practical to show all of the standard curves for each determination.	4	1	provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			life5010403_makarova	0
+p. 7, Figure 3, formate formula has been changed to “HCOO-” from “COOH-”.	2	1	It was particularly interesting to see the differences in protein functionality among the strains of marine Synechococcus.			life5010432_makarova	0
+p. 2, Lines 22–24; the data at the website cannot be posted as a supplementary figure because of copyright issues. I will ask them to revise the figure using English.	2	1	It was particularly interesting to see the differences in protein functionality among the strains of marine Synechococcus.			life5010432_makarova	0
+p. 2, Line 24; I confirmed that the following website link to the data of nitrite concentration is correct. “http://www.data.jma.go.jp/gmd/kaiyou/db/vessel_obs/hq/2006spr/137e/index_line.php?id=no2” 	2	1	entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			life5010432_makarova	0
+p. 4, line 16; Six NitM proteins registered recently were added to Figure 1 and Figure 5.	2	1	CC9605 is an open ocean strain and does not encounter high nitrite concentrations, while CC9311 is a coastal strain that deals with a range of nitrate/nitrite availability.			life5010432_makarova	0
+p5, Figure 1; Figure 1 was reproduced by using the UPGMA clustering method instead of the NJ clustering method according to the suggestion of the reviewer, then, NitM from α-cyanobacteria and β-cyanobacteria form clearly distinct groups. “using the UPGMA (Unweighted Pair Group Method with Arithmetic mean) clustering method of ClustalX.” was added in the legend of Figure 1.	2	1	entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			life5010432_makarova	0
+Formate inhibition experiments of the NitM from CC9311, CC9605 and MIT9313 have not been carried out yet.	2	1	Such differences can be explained by habitat adaption of each strain.			life5010432_makarova	0
+We will consider the nitrite uptake experiments at environmentally relevant concentrations of nitrite (~50 nM) in the future.	2	1	R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			life5010432_makarova	0
+p. 7, Figure 3, formate formula has been changed to “HCOO-” from “COOH-”.	2	1	entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			life5010432_perova	0
+p. 2, Lines 22–24; the data at the website cannot be posted as a supplementary figure because of copyright issues. I will ask them to revise the figure using English.	2	1	(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			life5010432_perova	0
+p. 2, Line 24; I confirmed that the following website link to the data of nitrite concentration is correct. “http://www.data.jma.go.jp/gmd/kaiyou/db/vessel_obs/hq/2006spr/137e/index_line.php?id=no2” 	2	1	Round 1: and Author Response I found this research interesting, relevant and novel.			life5010432_perova	0
+p. 4, line 16; Six NitM proteins registered recently were added to Figure 1 and Figure 5.	2	1	Such differences can be explained by habitat adaption of each strain.			life5010432_perova	0
+p5, Figure 1; Figure 1 was reproduced by using the UPGMA clustering method instead of the NJ clustering method according to the suggestion of the reviewer, then, NitM from α-cyanobacteria and β-cyanobacteria form clearly distinct groups. “using the UPGMA (Unweighted Pair Group Method with Arithmetic mean) clustering method of ClustalX.” was added in the legend of Figure 1.	2	1	Round 1: and Author Response I found this research interesting, relevant and novel.			life5010432_perova	0
+Formate inhibition experiments of the NitM from CC9311, CC9605 and MIT9313 have not been carried out yet.	2	1	Functional annotation of genes through protein expression is highly needed in our time of exponentially growing sequence data.			life5010432_perova	0
+We will consider the nitrite uptake experiments at environmentally relevant concentrations of nitrite (~50 nM) in the future.	2	1	Such differences can be explained by habitat adaption of each strain.			life5010432_perova	0
+Multi-alignment of concatenated protein (or DNA) segments is a genome-scale, but not whole-genome approach. Its applicability depends on the scope of the phylogenetic study. When dealing with not-too-distantly related species it may yield more or less useful result. However, in a study covering many phyla it is very difficult, if not impossible, to collect a common set of conserved proteins. Moreover, the concatenation method can never lead to very convincing conclusion, as give or take a few proteins may change the result. The phylogenomics people have noticed this problem, see, e.g., O. Jeffroy, H. Brinkman, F. Delsuc, H. Philippe (2008) Phylogenomics: the beginning of incongruence? Trends in Genetics, 22(4): 225–231. An example from the Bacteria domain is the relationship of the closely related Shigella and Escherichia coli strains. Concatenation of different number of genes led to different way of mixing-up of the two groups, but CVTree gave unambiguous separation of the strains as different species in the same genus Escherichia, see: G.-H. Zuo, Z. Xu, B.L. Hao (2013) Shigella strains are not clones of Escherichia coli but sister species in the genus Escherichia. Genomics Proteomics Bioinformatics, 11: 61–65. In order to carry out multi-alignment of concatenated sequences, a postdoc or well-trained PhD student equipped with the corresponding software is required. In contrast, with genome sequencing becoming a common practice in many labs it costs no additional work for a bench-microbiologist to get phylogenetic and taxonomic information by using a convenient and publically available tool such as the CVTree web serve. Well, we would be glad to see comparison of CVTree phylogeny with multi-alignment of concatenated proteins if anyone finds a way to do it for so many diverse phyla, but we do not consider it as a doable job.	2	1	Authors present a new interactive tree-visualization tool which enables direct validation of taxonomic groups according to their monophyly.			life5010949_makarova	0
+These points were discussed in the “Material and Method” section added at the suggestion of Reviewer 2. The following was copied from the manuscript: “Traditionally a newly generated phylogenetic tree is subject to statistical re-sampling tests such as bootstrap and jackknife. CVTree does not use sequence alignment. Consequently, there is no way to recognize informative or non-informative sites. Instead we take all the protein products encoded in a genome as a sampling pool for carrying out bootstrap or jackknife tests (citing our 2004 paper). Although it was very time-consuming, CVTrees did have well passed these tests (citing our 2010 paper). However, successfully passing statistical re-sampling tests only tells about the stability and self-consistency of the tree with respect to small variations of the input data. It is by far not a proof of objective correctness of the tree. Direct comparison of all branchings in a tree with an independent taxonomy at all ranks would provide such a proof, The 16S rRNA phylogeny cannot be verified by the Bergey's taxonomy, as the latter follows the former. However, agreement of branchings in CVTree with the Bergey's taxonomy would provide much stronger support to the tree as compared to statistical tests. This is the strategy we adopt for the CVTree approach.” “There are two aspects of a phylogenetic tree: the branching order (topology) and the branch lengths. Branching order is related to classification and branch length to evolution time. Calibration of branch lengths is always associated with the assumption that mutation rate R3 remains more or less a constant across all species represented in a tree, an assumption that cannot hold true in a large-scale phylogenetic study like the present one. Therefore, branching order in trees is of primary concern, whereas calibration of branch lengths makes less sense. Accordingly, all figures in this paper only show the branching scheme without indication of branch lengths and bootstrap values”.	2	1	Even if defined in the future, it must be lineage-dependent.			life5010949_makarova	0
+Yes, this is an apparent discrepancy of CVTree from 16S (and 23S) analysis for the given set of 179 archaeal genomes. However, in an on-going study of ours (not published yet) using a much larger data set this violation no longer shows up; both Korarchaeota and Crenarchaeota restore their phylum status. Taking into account the fact that both Korarchaeota and Thermofilaceae are represented by single species for the time being, their placement certainly requires further study with broader sampling of genomes.	2	1	Results also provide additional support to recently proposed archaeal phyla and halobacterial orders.			life5010949_makarova	0
+Highly degenerated genomes of many symbiont organisms tend to move around, in particular, to the baseline of a tree and thus distorts the overall structure of the tree. Therefore, it is better not to mix them with free-living organisms in a study. We rephrased the corresponding paragraph in the manuscript: “The nanosized archaean symbiont Nanoarchaeum equitans has a highly reduced genome (490,885 bp). It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota. We note that the monophyly of Euryarchaeota was also violated by Nanoarchaeum in some 16S rRNA trees, see, e.g., Figure 4 in a 2009 microbial survey as well as (c) and (d) in our Figure 3. It has been known that tiny genomes of endosymbiont microbes often tend to move towards baseline of a tree and distort the overall picture. In fact, we have suggested skipping such tiny genomes when studying bacterial phylogeny, see, e.g., (citing our 2010 paper) and a note in the home page of the CVTree Web Server. In the present case we may at most say that Nanoarchaeota probably makes a separate phylum, but its cutting into Euryarchaeota might be a side effect due to the tiny size of the highly reduced genome”.	2	1	present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+Yes, there was certain disturbing effect of the tiny and lonely Nanoarchaeum genome, yet the Halobacteria is a very specific clade, forming a tightly connected group and moving around as a whole, mainly due to the biased acidity of their constituent amino acids. We anticipate that the relative placement of Halobacteria with respect to other groups may stabilize when more genomes are used to construct a tree.	2	1	Even if defined in the future, it must be lineage-dependent.			life5010949_makarova	0
+We have A new “Material and Method” section has been added. Such issues as statistical resampling tests (bootstrap and jackknife), calibration of branch length, the meaning and choice of the peptide length K, etc. , were discussed in the new section. Figures 1 and 2 were combined to a new Figure 1; Figures 3 and 4 were combined to become a new Figure 2. Figure captions were made more detailed. The whole text was checked for language flaws and many places were rephrased.	2	1	Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			life5010949_makarova	0
+We thank the Reviewer for the detailed comments/suggestions given in the previous report and the suggestion of doing spelling-check this time. We have gone through the final manuscript carefully once more.	4	1	The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system.			life5010949_makarova	0
+A “Material and Method” section has been added where the CVTree algorithm, the interactive tree-viewer, statistical resampling tests (bootstrap, jackknife), calibration of branch lengths, etc., were discussed in slightly more detail.	2	1	are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			life5010949_makarova	0
+Yes, 16S rRNA phylogeny is quite stable and it almost defines the present taxonomy. We have given due credit for this. In general, CVTree does not challenge 16S rRNA analysis but complement it.	2	1	are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			life5010949_makarova	0
+A robust phylogenetic tree comes with a fixed branching order of leaves. One looks at the leaf names and their taxonomic lineage and tries to map the latter to the branches. To this end we added the following paragraphs in the “Material and Method” section. “There are two aspects of a phylogenetic tree: the branching order (topology) and the branch lengths. Branching order is related to classification and branch length to evolution time. Calibration of branch lengths is always associated with the assumption that mutation rate remains more or less a constant across all species represented in a tree, an assumption that cannot hold true in a large-scale phylogenetic study like the present one. Therefore, branching order in trees is of primary concern, whereas calibration of branch lengths makes less sense. Accordingly, all figures in this paper only show the branching scheme without indication of branch lengths and bootstrap values.” “Branching order in a tree by itself does not bring about taxonomic ranks, e,g, class or order.	2	1	However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			life5010949_makarova	0
+We have reorganized the manuscript mainly by adding a new “Material and Method” section where discussions on branch length, statistical resampling, meaning and choice of K, etc., were given. The original Figure 1 was deleted with some related points explained in the text accompanying the original Figure 2. All figure captions have been rewritten for clarity.	2	1	— Done Second Round of Evaluation Round 2: and Author Response			life5010949_makarova	0
+A few more sentences were added in the “Conclusion” regarding the power and achievement of the 16S rRNA analysis.	2	1	Results also provide additional support to recently proposed archaeal phyla and halobacterial orders.			life5010949_makarova	0
+We tried to rephrase the paragraph by changing, deleting, or adding a few words as follows: “In this paper we study Archaea phylogeny across many phyla. This is in contrast with phylogeny of species in a narrow range of taxa, e.g., that of vertebrates (a subphylum) or human versus close relatives (a few genera). Accordingly, the phylogeny should be compared with taxonomy at large, or, as Cavalier-Smith (citing cavalier-smith 2002) put it, with “megaclassificaton” of prokaryotes. Although in taxonomy the description of a newly discovered organism necessarily starts from the lower ranks, higher rank assignments are often incomplete or lacking. At present the ranks above class are not covered by the Bacteriological Code. The number of plausible microbial phyla may reach hundreds and archaeal ones are among the less studied. According to the 16S rRNA analysis, the major archaeal classes and their subordinate orders have been more or less delineated. Therefore, in order to carry out the aforementioned cross verification we make emphasis on higher ranks such as phyla, classes, and orders. A study using 179 Archaea genomes should provide a framework for further study of lower ranks.”	2	1	This kind of research should be encouraged further because old taxonomic paradigms must be systematically reviewed based on new genomic data.			life5010949_makarova	0
+Branching order in a tree is directly related to taxonomy, while branch lengths have more to do with evolution. For large-scale phylogenetic study across many phyla the former is more important than the calibration of branch lengths. The latter is based on the assumption that mutation rate is more or less constant. This assumption cannot hold when dealing with many phyla.	2	1	— Done Second Round of Evaluation Round 2: and Author Response			life5010949_makarova	0
+This is done in the newly added “Material and Method” section.	2	1	If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.”			life5010949_makarova	0
+The original Figure 1 was deleted and a few words added to the legend of the original Figure 2, now the new Figure 1.	2	1	Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			life5010949_makarova	0
+Judging by the cluster labeled as Euryarchaeote{0+3} in Figure 2 Methanomassiliicoccus was not reclassified into Thermoplasmataceae but to an yet un-specified class.	2	1	Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+The sentence has been moved to the legend of Fig. 1 and slightly rephrased.	2	1	are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			life5010949_makarova	0
+Done.	2	1	Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+the K issue is discussed in the newly added “Material and Method” section; so scattered mentioning of K has been deleted from the rest of text.	2	1	For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			life5010949_makarova	0
+Done in the caption of Figure 2.	2	1	General Comments The topic is relevant for microbial taxonomy.			life5010949_makarova	0
+We have rewritten the paragraph as: “The new phylum Korarchaeota violates the monophyly of the phylum Crenarchaeota by drawing to itself the family Thermofilaceae. However, in an on-going study of ours (not published yet) using a much larger data set, this violation no longer shows up; both R8 Korarchaeota and Crenarchaeota restore their phylum status. Taking into account the fact that both Korarchaeota and Thermofilaceae are represented by single species for the time being, their placement certainly requires further study with broader sampling of genomes.”	2	1	It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			life5010949_makarova	0
+From the original Figures 3 and 4 only one has been kept and the legend rewritten. In fact, the whole paragraph changed to: “The newly proposed phylum Thaumarchaeota appears to be non-monophyletic as an outlying strain Candidatus Caldiarchaeum subterranum was assigned to this phylum according to the NCBI taxonomy. The NCBI assignment might reflect its position in some phylogenetic tree based on concatenated proteins, e.g., Figure 2 in […]. However, in the original paper reporting the discovery of this strain […] and in recent 16S rRNA studies, e.g., […], Candidatus Caldiarchaeum subterranum was proposed to make a new phylum Aigarchaeota. CVTrees support the introduction of this new phylum. A lineage modification of Candidatus Caldiarchaeum subterranum from Thaumarchaeota to Aigarchaeota would lead to a monophyletic Thaumarchaeota{7}.”	2	1	R13 Round 2: and Author Response Authors have substantially improved the article, including language corrections, and have provided extensive clarifications to all initial criticisms.			life5010949_makarova	0
+The problem of taxonomic placement of Aciduliprofundum is a good example to demonstrate how one extract information from CVTrees. In the Reysenbach et al. Nature 2006 paper it was taken as the first cultivated member of the DHEV2 (deep-sea hydrothermal euryarchaeate 2) clade based on a maximum-likelihood 16S rRNA tree. Unfortunately, all other 13 members of this clade were represented by 16S rRNA sequences only and no genome data are available so far. The NCBI taxonomy gave an incomplete lineage: Archaea; Euryrchaeota; unclassified Euryarchaeota; missing taxonomic assignment at the rank class and below. In order to make use of CVTree we must touch on the K-issue a little more. The alignment-free comparison of genomes in CVTree is implemented by counting the number of K-peptides in the protein products encoded in a genome followed by subtraction of a random background caused by neutral mutations. The peptide length K looks like a parameter, but it is actually not a parameter. Using a longer K emphasizes species-specificity, while a shorter K takes into account more common features with neighboring species. However, we never adjust K: a fixed K is used for all genomes to construct a tree, but one may construct a series of trees for K = 3, 4, 5, 6, 7, 8, … We have shown repeatedly that R9 K = 5 and 6 lead to best results in the sense of agreement with taxonomy, so usually only a K = 6 tree is given in publications. Let us look at a subtree, i.e., part of a tree, containing the organisms of interest. If the branching order in all trees built for different Ks turns out to be same, it would be a strong support to the branching order. In most cases the branching order varies with K: K = 3 and 4 make sense, K = 5 and 6 yield the best, K = 7 and 8 become slightly worse, etc. For too big a K, even if the closest strains remain grouped together the whole tree may tend to become a star-tree, i.e., every small clade stands in its own and their mutual placements become less meaningful. Therefore, inspection of trees for a range of K-values provides an additional dimension to evaluate the results. For Aciduprofumdum we have a stable pair (<C>Thermococci{18}, <G>Aciduprofumdum{2}) at K=3, 5, 6, 7. At K = 4 we have (<C>Thermococci{18}, (<G>Staphylothermus{2}, <G>Aciduprofundum{2})) In all these cases Thermoplasmata stands farther away from the above pair. However, at K = 8 and 9, when the overall tree picture has been largely distorted, Aciduliprofundum does stand closer to Thermaplasmata. Putting together all the above results we tend to consider the pair (<C>Thermococci{18}, <G>Aciduliprofundum{2}) as reflecting a more probable relation. Confined to the available data for the time being one may assign <G>Aciduliprofundum to <C>Thermococci, e.g., to denote the pair as <C>Thermococci{20}=(<F>Thermococcaceae{18}, <G>Aciduliprofundum{2}) leaving its family unclassified or assign it to a new family. Without further phenotypic and chemotaxonomic evidence it is better not to introduce new taxon names if the present naming scheme is capable to accommodate the leaves without conflict. This was why we wrote “this modification would hold as long as no new facts challenge it”. Anyway, taxonomy has always been a work in progress. One has to be prepared for modifications when new data appear. To make a long story short, we have rewritten the paragraph as: “The Candidatus genus Aciduliprofundum is considered a member of the DHEV2 (deap-sea hydrothermal vent euryarchaeotic 2) phylogenetic cluster. No taxonomic information was given in the original papers [55,56]. The NCBI Taxonomy did not provide definite lineage information for this taxon at the class, order, and family ranks. According to [55] the whole DHEV2 cluster was located close to Thermoplamatales in a maximum-likelihood analysis of 16S rRNA sequences. A similar placement was seen in [54] where a Bayesian tree of the archaeal domain based on concatenation of 57 ribosomal proteins put a lonely Aciduliprofundum next to Thermoplasmata. However, in CVTrees, constructed for all K-values from 3 to 9, Aciduliprofundum juxstaposes with the class Thermococci{18}. An observation in [56] that this organism shares a rare lipid structure with a few species from Thermococcales may hint on its possible association with the latter. If we temporarily presume a lineage <C>Thermococci<O>Unclassified<F>Unclassified<G>Aciduliprofundum… R10 one might have a monophyletic class <C>Thermococci{20}. Since none of the 13 DHEV2 members listed in [55] has a sequenced genome so far, CVTree cannot tell the placement of the DHEV2 cluster as a whole for the time being. It remains an open problem as whether DHEV2 is close to Thermoplasmata or to Thermococci, or a new class is needed to accommodate DHEV2.”	2	1	General Comments The topic is relevant for microbial taxonomy.			life5010949_makarova	0
+No, we did not mean it.	2	1	— Done Second Round of Evaluation Round 2: and Author Response			life5010949_makarova	0
+We should first explain how these inappropriate names appeared. We have insisted to use the directory name at the NCBI FTP site as genome name. However, in November 2013 NCBI announced that they would not release genomes of different strains of the same species as before. In a period thereafter NCBI sometimes put several genomes in a directory and we had to extract the data and to assign a name from the “Source” line of the GenBank file. This caused some confusion. For example, as of February 27, 2015, a directory name at NCBI remained “archaeon_Mx1201_uid196597” and we had to change it to: Candidatus_Methanomethylophilus_alvus_Mx1201_uid196597 Now all “wrong names” as pointed out by the Reviewer no longer appear in figures. In the text we tried to refer to their names as complete as possible.	2	1	However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			life5010949_makarova	0
+The genome name at NCBI FTP site is “halophilic_archaeon_DL31_uid72619”. The uid number was dropped when mentioned in the text. We put it back and capitalized the first letter to “Halophilic”, still an illegal genus name.	2	1	If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.”			life5010949_makarova	0
+It is Figure 2 in the revised manuscript. We discussed it at some length.	2	1	Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+No, organism cannot be published. Thanks for correcting our mistake.	2	1	Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system. Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being. Even if defined in the future, it must be lineage-dependent. For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla. In addition, monophyly is a guiding principle in comparing branching order with taxonomy. Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy. If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.” R6 One cannot tell the rank of a node/leaf in a tree by simply looking at it. A reference taxonomy is alwys needed. We put the following in the “Material and Method” section to explain it: “Branching order in a tree by itself does not bring about taxonomic ranks, e,g, class or order.	2	1	Authors emend classification of some strains.			life5010949_makarova	0
+The phrase “3063 identical nucleotide positions” was copied from the caption of Figure 4 of the cited Nunoura et al. 2011 paper without much thinking. We simply deleted it.	2	1	About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			life5010949_makarova	0
+The whole paragraph has been rewritten as: “The nanosized archaean symbiont Nanoarchaeum equitans has a highly reduced genome (490,885 bp [44]). We note that the monophyly of Euryarchaeota was also violated by Nanoarchaeum in some 16S rRNA trees, see, e.g., Figure 4 in a 2009 paper [61] as well as (c) and (d) in our Figure 4. In fact, we have suggested skipping such tiny genomes when studying bacterial phylogeny, see, e.g., [29] and a note in the home page of the CVTree Web Server [21]. In the present case we may at most say that Nanoarchaeota probably makes a separate phylum, but its cutting into Euryarchaeota might be a side effect due to the tiny size of the highly reduced genome.”	2	1	However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			life5010949_makarova	0
+As a method CVTree is independent of 16S rRNA analysis. First, it uses protein products in a genome instead of RNA segments in the genome. Second, it does not do sequence alignment. CVTree generates stable trees but cannot tell which branch corresponds to what taxon. Only after comparison with the existing classification and nomenclature one would be able to make connections with taxonomy. In this sense it does depend on 16S rRNA taxonomy. Anyway, CVTree does not challenge 16S rRNA analysis but makes it more convincing in most cases. The revealed discrepancies call for further study.	2	1	are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			life5010949_makarova	0
+In fact, genomic approaches are more effective at species level and below due to their high resolution power. At high ranks CVTree may be more effective in the sense that it does not require additional work. Suffice it to put genomes in CVTree web server and the branches come out, then compare them with a reference taxonomy.	2	1	present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+Even “herenow” does not seem to be an correct English word; we changed it to “so far”.	2	1	Authors present a new interactive tree-visualization tool which enables direct validation of taxonomic groups according to their monophyly.			life5010949_makarova	0
+We changed “is in contrast with” to “is distinct from” and added a phrase “focusing on taxonomy of higher ranks” at the end of a sentence. Now the sentences read: “This is distinct from phylogeny of species in a narrow range of taxa, e.g., that of vertebrates (a subphylum) or human versus close relatives (a few genera). Accordingly, the phylogeny should be compared with taxonomy at large, or, as Cavalier-Smith \cite{cavalier-smith2002} put it,with “megaclassificaton” of prokaryotes, focusing on taxonomy of higher ranks.” -	4	1	— Done Second Round of Evaluation Round 2: and Author Response			life5010949_makarova	0
+done	4	1	For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			life5010949_makarova	0
+Moved to the conclusion section and the first word “Though” replaced by “In addition, since”	4	1	Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+The whole paragraph has been deleted. This paragraph was added to the revised manuscript because one of the Reviewers asked “Does CVTree still require input genome data to be annotated to gene features, i.e., protein or CDS?” Well, this question reminds us that for many so-called “Permanent Draft” genomes it may be worthwhile returning to our early practice of using whole genome nucleotide sequences without distinguishing coding and non-coding segments. Although it did not lead to better results as compared with using translated protein products, but it is doable on un-annotated contigs. We will try this later.	4	1	About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			life5010949_makarova	0
+We have deleted all what appeared within the curly brackets and kept only “monophyletic Thaumarchaeota” as suggested. In fact, we could not tell how these words appeared there; there was none in our draft manuscript.	4	1	The tree was compared with several 16S rRNA trees reported in the literature, and the differences were minor.			life5010949_makarova	0
+Bending modulus is a material constant which does not depend on the size and shape of the object, but on the composition of the vesicle membrane and aqueous solution used (Lines 121–123 in the new version). We have inserted a definition of the membrane bending modulus (Lines 115–116) while the text regarding this issue extends between Lines 112–128.	2	1	Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			life5021101_makarova	0
+As the referee suggested The Theory and Experimental section were organized into the Section “Materials and Methods”. Other sections were renumbered accordingly.	2	1	According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			life5021101_makarova	0
+As it was explained in the text our experimental method is developed for application to almost spherical vesicles. Vesicles with diameters between 20 and 40 micrometers were considered, whose deviations from spherical shape (fluctuation of the radius) were small in comparison with the mean sphere radius. We added the information concerning the shape and the size of the studied objects and added pictures of the equatorial cross section of a fluctuating vesicle (new Figure 3), as seen under the phase contrast microscope.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_makarova	0
+We have rewritten the introductory paragraph (Lines 46–60).	2	1	Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_makarova	0
+Thank you for the comment, there was a mistake. We have tidied and updated the references.	2	1	According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			life5021101_makarova	0
+We thank the reviewer for further comments and hope that we can clarify the issues as given below. We have made changes in the manuscript (marked red) and added one new reference (ref. [35]).	4	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_makarova	0
+In principal in order to obtain elastic constants you need to apply force and measure the deformation that this force causes. If we want to obtain the bending elasticity of lipid vesicle we need a force with a very small power. In the case of thermally induced shape fluctuations as a force we use the Brownian (thermal) motion of water molecules, bombarding the membrane. This force is stochastic (we do not know its instant value), but the mean value of it is proportional to the temperature. As a result of this stochastic bombardment the lipid vesicle deforms (changes its shape or fluctuates). We acquire a big amount of pictures of fluctuating vesicle in order to get the mean value of the deformation that our force causes. To extract mean we need to have stationary conditions over the time, so the mean is taken, that is why it is really important to have constant temperature throughout the experiment.	4	1	Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			life5021101_makarova	0
+Using our experimental system we can measure bending elasticity also at higher temperatures (up to approximately 40 degrees C).	4	1	According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			life5021101_makarova	0
+The bending elasticity modulus depends on the temperature below and near the phase transition temperature, but far above the phase transition temperature (this is the case in our experiment) the bending elasticity modulus is practically constant. See: Temperature and Chain Length Effects on Bending Elasticity of Phosphatidylcholine Bilayers. Fernandez-Puente, I. Bivas, M. D. Mitov and P. Meleard, Europhys. Lett., 28, 181 (1994). We have added this text to the Discussion (Lines 234–237) and also added the reference to the reference list (ref.	4	1	Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			life5021101_makarova	0
+It is not a problem for us to make pictures of a fluctuating vesicle at different temperatures, but these photos would not be related to the manuscript.	4	1	Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_makarova	0
+We thank the referee for pointing to negligent mistakes in numbering. We have corrected wrong numbering of subsections.	4	1	Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			life5021101_makarova	0
+As the referee suggested, we have added data for the bending elasticity modulus of another lipid type—POPC lipid membrane. Also, we have added a new figure (Figure 3).	2	1	Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_makarova	0
+A number of improvements of the experimental procedure for the thermally induced shape fluctuation method were used in this study: the stroboscopic illumination was improved on several steps to achieve a better experimental conditions; a thermostatic stage was used to set and control a constant temperature, the analysing procedure was improved by adding strict objective criteria for qualification of the vesicle as a whole as well as for acceptance or rejection of a given contour of the sequence of recorded images and the white noise contribution to the amplitudes of thermal shape fluctuations was R3 evaluated and taken into account. We have given short description of the improvements made and for each of them cited the corresponding articles for detailed explanation. (Lines 181–184; 187–193).	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_makarova	0
+SOPC was chosen as both lipid types have transition temperature far below the temperature at experiments. In the revised version, we measured the bending modulus for another lipid type (POPC), measured and analysed by the same method for the same experimental conditions.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_makarova	0
+We have corrected the wrong number.	2	1	Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			life5021101_makarova	0
+The aim of the present study was to obtain the bending elasticity modulus of the archaeal membrane at fixed temperature (27 degrees C) far above the phase transition temperature. We have not measured the temperature dependence of the bending constant.	2	1	According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			life5021101_makarova	0
+In this work we present the bending elasticity modulus of archaeal lipid membrane in pure water environment. We added the details concerning the water purification (Lines 173–175). In other works the influence of different admixtures in the aqueous solution around the membrane was studied and the values depending on the type and the concentration of it for given lipid were reported. As the referee suggested we added such information with the corresponding references in the introduction section (Lines 57–60).	2	1	Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			life5021101_makarova	0
+As I understand the referee had no further comments.	4	1	Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			life5021101_makarova	0
+Bending modulus is a material constant which does not depend on the size and shape of the object, but on the composition of the vesicle membrane and aqueous solution used (Lines 121–123 in the new version). We have inserted a definition of the membrane bending modulus (Lines 115–116) while the text regarding this issue extends between Lines 112–128.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+As the referee suggested The Theory and Experimental section were organized into the Section “Materials and Methods”. Other sections were renumbered accordingly.	2	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+As it was explained in the text our experimental method is developed for application to almost spherical vesicles. Vesicles with diameters between 20 and 40 micrometers were considered, whose deviations from spherical shape (fluctuation of the radius) were small in comparison with the mean sphere radius. We added the information concerning the shape and the size of the studied objects and added pictures of the equatorial cross section of a fluctuating vesicle (new Figure 3), as seen under the phase contrast microscope.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+It would also be helpful to clarify/expand the description about the drug delivery system using the archaeal vesicle. The first sentence in the introduction section should be corrected as cell should not be considered to be “the building block of life”. It’s better to re-write the “basic motivation” for the study.	2	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+Thank you for the comment, there was a mistake. We have tidied and updated the references.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+We thank the reviewer for further comments and hope that we can clarify the issues as given below. We have made changes in the manuscript (marked red) and added one new reference (ref. [35]).	4	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+In principal in order to obtain elastic constants you need to apply force and measure the deformation that this force causes. If we want to obtain the bending elasticity of lipid vesicle we need a force with a very small power. In the case of thermally induced shape fluctuations as a force we use the Brownian (thermal) motion of water molecules, bombarding the membrane. This force is stochastic (we do not know its instant value), but the mean value of it is proportional to the temperature. As a result of this stochastic bombardment the lipid vesicle deforms (changes its shape or fluctuates). We acquire a big amount of pictures of fluctuating vesicle in order to get the mean value of the deformation that our force causes. To extract mean we need to have stationary conditions over the time, so the mean is taken, that is why it is really important to have constant temperature throughout the experiment.	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+Using our experimental system we can measure bending elasticity also at higher temperatures (up to approximately 40 degrees C).	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+The bending elasticity modulus depends on the temperature below and near the phase transition temperature, but far above the phase transition temperature (this is the case in our experiment) the bending elasticity modulus is practically constant. See: Temperature and Chain Length Effects on Bending Elasticity of Phosphatidylcholine Bilayers. Fernandez-Puente, I. Bivas, M. D. Mitov and P. Meleard, Europhys. Lett., 28, 181 (1994). We have added this text to the Discussion (Lines 234–237) and also added the reference to the reference list (ref.	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+It is not a problem for us to make pictures of a fluctuating vesicle at different temperatures, but these photos would not be related to the manuscript.	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+We thank the referee for pointing to negligent mistakes in numbering. We have corrected wrong numbering of subsections.	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+As the referee suggested, we have added data for the bending elasticity modulus of another lipid type—POPC lipid membrane. Also, we have added a new figure (Figure 3).	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+A number of improvements of the experimental procedure for the thermally induced shape fluctuation method were used in this study: the stroboscopic illumination was improved on several steps to achieve a better experimental conditions; a thermostatic stage was used to set and control a constant temperature, the analysing procedure was improved by adding strict objective criteria for qualification of the vesicle as a whole as well as for acceptance or rejection of a given contour of the sequence of recorded images and the white noise contribution to the amplitudes of thermal shape fluctuations was R3 evaluated and taken into account. We have given short description of the improvements made and for each of them cited the corresponding articles for detailed explanation. (Lines 181–184; 187–193).	2	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+SOPC was chosen as both lipid types have transition temperature far below the temperature at experiments. In the revised version, we measured the bending modulus for another lipid type (POPC), measured and analysed by the same method for the same experimental conditions.	2	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+We have corrected the wrong number.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+The aim of the present study was to obtain the bending elasticity modulus of the archaeal membrane at fixed temperature (27 degrees C) far above the phase transition temperature. We have not measured the temperature dependence of the bending constant.	2	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+In this work we present the bending elasticity modulus of archaeal lipid membrane in pure water environment. We added the details concerning the water purification (Lines 173–175). In other works the influence of different admixtures in the aqueous solution around the membrane was studied and the values depending on the type and the concentration of it for given lipid were reported. As the referee suggested we added such information with the corresponding references in the introduction section (Lines 57–60).	2	1	The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+As I understand the referee had no further comments.	4	1	However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+We have rephrased the four paragraphs where the referee found that the described examples are not always easy to follow.	2	1	R2 Round 1: and Author Response I received this manuscript and looked at it with joy.			life5021427_makarova	0
+We have added a short “outlook-type” paragraph towards the end of the conclusions. In this paragraph we mention three approaches that will facilitate gene discovery, among them large-scale screening techniques as exemplified by the insertion mutant library pointed out by the referee.	2	1	Importance: Archaea is/are the third domain of life, no more or less!			life5021427_makarova	0
+We have rephrased the four paragraphs where the referee found that the described examples are not always easy to follow.	2	1	The authors describe an alternative, manual curation strategy aimed at avoiding over-annotation.			life5021427_perova	0
+We have added a short “outlook-type” paragraph towards the end of the conclusions. In this paragraph we mention three approaches that will facilitate gene discovery, among them large-scale screening techniques as exemplified by the insertion mutant library pointed out by the referee.	2	1	The curation procedure is done, to my mind, very well; and I am confident that this will help the archaea community enormously.			life5021427_perova	0
+The authors would like to thank for this helpful comment. We changed the subsection as described below. With stating that argon-nitrogen mixtures mainly emit high amounts of radiation, the authors want to emphasize, that the sterilization efficiency is based on radiation rather than interaction of radicals with the sample. In low-pressure argon-nitrogen plasmas, the only significant possible radical is atomic nitrogen. As atomic nitrogen is not known for high etching efficiency towards organic materials, the main reason for inactivation of D. radiodurans is based on radiation effects. Furthermore, Argon-Nitrogen plasmas emit high amounts of radiation compared to Argon-Oxygen plasmas, as shown for example in [5]. The data in [5] is not comparable in absolute values, as the system was changed afterwards. Absolutely calibrated spectra were measured and have been submitted in another publication. The authors think that estimating the inactivation efficiency towards D. radiodurans based on the UV dose cannot be performed on a reliable bases due to the following reasons: First, the inactivation efficiency towards spores is not only depending on the overall dose of the emitted radiation, but especially on the dose per wavelength. This has been demonstrated by Munakata et al. for B. subtilis spores (Munakata et al., Photochemistry and Photobiology, Blackwell Publishing Ltd, 1991, 54, 761-768). To the authors’ knowledge, there is no publication measuring the wavelength depending inactivation efficiency of B. radiodurans in the VUV. Second, the samples are mixed with Mars Analog Soil and most of the samples are covered with it. This leads to a very inhomogeneous exposure of the samples which can only be estimated arbitrarily. To address the reviewer’s comments, subsection 2.3 has been modified as follows: An argon-nitrogen mixture (100:5 sccm) was used with continuous power of 500 W, as it emits significantly higher amounts of radiation in the bactericidal wavelength region from 100 nm to 400 nm compared to an argon-oxygen mixture (100:5 sccm) [5]. The absolute values presented in [5] are not applicable to the results shown here, due to modification of the plasma system concerning the power coupling. Nevertheless, the significantly higher emission of the argon-nitrogen mixture is shown. Absolute values of the radiation dose in the range from 100 nm to 400 nm of the system configuration used in this study were measured and submitted [6]. A possible radical that can interact with the samples is atomic nitrogen. As atomic nitrogen is not known for etching of organic materials, we assume that inactivation in the argon-nitrogen mixture is based on radiation effects. [6] Raguse, M.; Fiebrandt, M.; Denis, B.; Stapelmann, K.; Eichenberger, P.; Driks, A.; Eaton, P.; Awakowicz, P.; Moeller, R. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low pressure plasma sterilization. Submitted to Journal of Physics D: Applied Physics.	2	1	Line 50: R1T (=ATCC 13939T=DSM 20539T) instead of R1 ATCC13939/DSM20539 Line 57: Are you sure that the Martian Analog soils were sterile before using?			life6020022_makarova	0
+The authors thank the referee for this comment that demonstrates that the authors need to clarify. However, to the authors it is not clear which potentially inactivating components of the plasma are not specified, as radicals and radiation are mentioned and the temperature is kept below 80°C to prevent inactivation effects due to heating. As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV. Thus, sputtering of the sample due to ion bombardment can be neglected. Low-pressure plasmas dissociate molecular gases up to several percent. Thus, the authors wanted to emphasize, that the atomic oxygen density and therewith the atomic oxygen flux to the samples is in the order of a few percent of the overall gas flux. Quantitative and even qualitative analysis of atomic oxygen densities by optical emission spectroscopy is not an easy task as several parameters like gas temperature, electron density and electron temperature are necessary. This is even more challenging in pulsed mode, as the plasma does not achieve a steady state due to switching it on and off to prevent intensive heating of the plasma. Determination of the gas densities and plasma parameters is currently addressed in extensive analysis of plasmas with different gas compositions but not finished and published yet. Nevertheless, the authors’ state, that a relevant amount of atomic oxygen species is produced in the low pressure argon-oxygen plasma as the minimum energy for dissociation of molecular oxygen is around 5 eV. The ionization energy for argon is 16 eV and 13 eV for molecular oxygen. Thus, sufficient electrons with enough energy to dissociate molecular oxygen will be present in the plasma for dissociation up to a few percent. In the authors’ view, estimating the mean free path of the atomic oxygen in the plasma is not necessary, as recombination of radicals in low pressure plasmas in the regime of a few Pascal takes place at the surfaces. Thus, any radical formed will be lost at the walls and not on its way to a surface. Estimating the mean free path of the radical in the Mars Analog Soil can only be performed on an arbitrary basis as the sticking and recombination coefficients of atomic oxygen are not known for the Mars Analog Soil. Thus, any calculation of the mean free path of the radical is based on estimated data and could only be verified by experiments. To address the reviewer’s comments, subsection 2.3 has been modified as follows: Only JSC Mars-1A analog soil was used in the second trial. In this case, an argon-oxygen mixture (100:5 sccm) was used as it produces reactive oxygen species due to dissociation of molecular oxygen in the plasma. Since radiation-based sterilization is faster than sterilization due to oxidation, the treatment time was increased. As this leads to heating of the sample above 80 °C in the continuous plasma, we used a pulsed mode, switching the plasma on and off at a frequency of 1 kHz and a duty cycle of 10 %. To account for this fact, the treatment time was increased to 45 min, yielding a plasma exposure time of 4.5 min. Furthermore, the power was increased to 1500 W, yielding a mean power of 150 W, as it is only applied 10 % of the time. The plasma conditions are not matched to each other as the main objective was to observe to what degree the cells of D. radiodurans were affected by radiation or reactive species and not which component is more efficient. Thus, sputtering of the sample due to ion bombardment can be neglected as possible inactivation mechanism.	2	1	Minor comments Lines 31-32: Bacillus subtilis and B.pumilus should be written in italic.			life6020022_makarova	0
+The grain size distributions of the three Mars regolith simulants have been added to the text in form of two additional tables (table 2: P-MRS and S-MRS; and table 3: JSC Mars-1A), as well as a new citation for the grain size distribution of JSC Mars-1A [8].	2	1	Line 78: Please explain in the text what is “a vacuum treatment control”.			life6020022_makarova	0
+To assess the shading effect of the regolith is difficult. As described we used a very thin layer of regolith particles, but did not measure them according to their shading effect but according to the weight, which was the same for all types of regolith. This experiment was meant as a test of the sterilization method adjusted to our purposes, as described in the introduction section. Therefore, we did not plan it according to the maximal theoretically achievable sterilization effect, but to design a life detection mission to Mars. [8] Wan, L., Wendner, R., & Cusatis, G. A Novel Material for In Situ Construction on Mars: Experiments and Numerical Simulations. 2016, arXiv preprint arXiv:1512.05461	2	1	Line 78: Please explain in the text what is “a vacuum treatment control”.			life6020022_makarova	0
+The title has been changed.	2	1	With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.			life6020022_makarova	0
+In the introduction we already described the problem of the destructiveness of heat sterilization like autoclaving on the organic contents of a soil, which was our motivation to test an alternative method such as low temperature plasma sterilization. D. radiodurans was used as test organism because of its resistance towards UV radiation, which is a part of the sterilizing effect of plasma sterilization. The appropriate passages in the introduction section have been edited.	2	1	With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.			life6020022_makarova	0
+Since we did not intend to test the influences of the Martian Analog soils on D. radiodurans, but the sterilizing effect of the low temperature plasma treatment on cells intermixed with soil, we did not make a control to determine the effects of the Mars analog soils. The starting values of cell numbers were estimated on samples prepared the same way as the samples for the plasma sterilization (intermixed with soils and desiccated as described in the method section). They were incubated on TYG agar plates like the plasma samples after the treatment. This explanation was missing in the text and has now been added to the appropriate passages in the method section (Chapter 2.2.) for a better understanding.	2	1	Line 78: Please explain in the text what is “a vacuum treatment control”.			life6020022_makarova	0
+The names are written in italics now, the mistake must have happened during the editing of the text.	2	1	First of all it is necessary to change the title and at least remove “The testing”.			life6020022_makarova	0
+Done, thank you.	2	1	The manuscript contains scientific material that can be published as a short communication.			life6020022_makarova	0
+Yes, we just used one medium (TYG), so we changed it to ‘medium’.	2	1	The mineralogical composition is given in table, but not the grain size distribution.			life6020022_makarova	0
+The writing has been changed accordingly.	2	1	Comments: The plasma conditions are not described sufficiently.			life6020022_makarova	0
+Since the bottles containing the Martian Analog soils were autoclaved before use, they should be sterile. We also made a separate test to check for sterility after autoclaving the Martian Analog soils, which has been negative (meaning no contamination could be detected via plating of the autoclaved JSC Mars-1A regolith on TYG medium).	2	1	In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.			life6020022_makarova	0
+An explanation has been added to the text passage.	2	1	Minor comments Lines 31-32: Bacillus subtilis and B.pumilus should be written in italic.			life6020022_makarova	0
+The differences might be caused due to the finer grain size of the other regolith used - particularly P-MRS, which possibly increased the shielding effect. This has been added to the discussion section.	2	1	Author Response Reviwers comments The manuscript contains scientific material that can be published as a short communication.			life6020022_makarova	0
+The authors would like to thank for this helpful comment. We changed the subsection as described below. With stating that argon-nitrogen mixtures mainly emit high amounts of radiation, the authors want to emphasize, that the sterilization efficiency is based on radiation rather than interaction of radicals with the sample. In low-pressure argon-nitrogen plasmas, the only significant possible radical is atomic nitrogen. As atomic nitrogen is not known for high etching efficiency towards organic materials, the main reason for inactivation of D. radiodurans is based on radiation effects. Furthermore, Argon-Nitrogen plasmas emit high amounts of radiation compared to Argon-Oxygen plasmas, as shown for example in [5]. The data in [5] is not comparable in absolute values, as the system was changed afterwards. Absolutely calibrated spectra were measured and have been submitted in another publication. The authors think that estimating the inactivation efficiency towards D. radiodurans based on the UV dose cannot be performed on a reliable bases due to the following reasons: First, the inactivation efficiency towards spores is not only depending on the overall dose of the emitted radiation, but especially on the dose per wavelength. This has been demonstrated by Munakata et al. for B. subtilis spores (Munakata et al., Photochemistry and Photobiology, Blackwell Publishing Ltd, 1991, 54, 761-768). To the authors’ knowledge, there is no publication measuring the wavelength depending inactivation efficiency of B. radiodurans in the VUV. Second, the samples are mixed with Mars Analog Soil and most of the samples are covered with it. This leads to a very inhomogeneous exposure of the samples which can only be estimated arbitrarily. To address the reviewer’s comments, subsection 2.3 has been modified as follows: An argon-nitrogen mixture (100:5 sccm) was used with continuous power of 500 W, as it emits significantly higher amounts of radiation in the bactericidal wavelength region from 100 nm to 400 nm compared to an argon-oxygen mixture (100:5 sccm) [5]. The absolute values presented in [5] are not applicable to the results shown here, due to modification of the plasma system concerning the power coupling. Nevertheless, the significantly higher emission of the argon-nitrogen mixture is shown. Absolute values of the radiation dose in the range from 100 nm to 400 nm of the system configuration used in this study were measured and submitted [6]. A possible radical that can interact with the samples is atomic nitrogen. As atomic nitrogen is not known for etching of organic materials, we assume that inactivation in the argon-nitrogen mixture is based on radiation effects. [6] Raguse, M.; Fiebrandt, M.; Denis, B.; Stapelmann, K.; Eichenberger, P.; Driks, A.; Eaton, P.; Awakowicz, P.; Moeller, R. Understanding of the importance of the spore coat structure and pigmentation in the Bacillus subtilis spore resistance to low pressure plasma sterilization. Submitted to Journal of Physics D: Applied Physics.	2	1	The manuscript contains scientific material that can be published as a short communication.			life6020022_perova	0
+The authors thank the referee for this comment that demonstrates that the authors need to clarify. However, to the authors it is not clear which potentially inactivating components of the plasma are not specified, as radicals and radiation are mentioned and the temperature is kept below 80°C to prevent inactivation effects due to heating. As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV. Thus, sputtering of the sample due to ion bombardment can be neglected. Low-pressure plasmas dissociate molecular gases up to several percent. Thus, the authors wanted to emphasize, that the atomic oxygen density and therewith the atomic oxygen flux to the samples is in the order of a few percent of the overall gas flux. Quantitative and even qualitative analysis of atomic oxygen densities by optical emission spectroscopy is not an easy task as several parameters like gas temperature, electron density and electron temperature are necessary. This is even more challenging in pulsed mode, as the plasma does not achieve a steady state due to switching it on and off to prevent intensive heating of the plasma. Determination of the gas densities and plasma parameters is currently addressed in extensive analysis of plasmas with different gas compositions but not finished and published yet. Nevertheless, the authors’ state, that a relevant amount of atomic oxygen species is produced in the low pressure argon-oxygen plasma as the minimum energy for dissociation of molecular oxygen is around 5 eV. The ionization energy for argon is 16 eV and 13 eV for molecular oxygen. Thus, sufficient electrons with enough energy to dissociate molecular oxygen will be present in the plasma for dissociation up to a few percent. In the authors’ view, estimating the mean free path of the atomic oxygen in the plasma is not necessary, as recombination of radicals in low pressure plasmas in the regime of a few Pascal takes place at the surfaces. Thus, any radical formed will be lost at the walls and not on its way to a surface. Estimating the mean free path of the radical in the Mars Analog Soil can only be performed on an arbitrary basis as the sticking and recombination coefficients of atomic oxygen are not known for the Mars Analog Soil. Thus, any calculation of the mean free path of the radical is based on estimated data and could only be verified by experiments. To address the reviewer’s comments, subsection 2.3 has been modified as follows: Only JSC Mars-1A analog soil was used in the second trial. In this case, an argon-oxygen mixture (100:5 sccm) was used as it produces reactive oxygen species due to dissociation of molecular oxygen in the plasma. Since radiation-based sterilization is faster than sterilization due to oxidation, the treatment time was increased. As this leads to heating of the sample above 80 °C in the continuous plasma, we used a pulsed mode, switching the plasma on and off at a frequency of 1 kHz and a duty cycle of 10 %. To account for this fact, the treatment time was increased to 45 min, yielding a plasma exposure time of 4.5 min. Furthermore, the power was increased to 1500 W, yielding a mean power of 150 W, as it is only applied 10 % of the time. The plasma conditions are not matched to each other as the main objective was to observe to what degree the cells of D. radiodurans were affected by radiation or reactive species and not which component is more efficient. Thus, sputtering of the sample due to ion bombardment can be neglected as possible inactivation mechanism.	2	1	Minor comments (please see changes in text at the relevant lines)			life6020022_perova	0
+The grain size distributions of the three Mars regolith simulants have been added to the text in form of two additional tables (table 2: P-MRS and S-MRS; and table 3: JSC Mars-1A), as well as a new citation for the grain size distribution of JSC Mars-1A [8]. To assess the shading effect of the regolith is difficult. As described we used a very thin layer of regolith particles, but did not measure them according to their shading effect but according to the weight, which was the same for all types of regolith. This experiment was meant as a test of the sterilization method adjusted to our purposes, as described in the introduction section. Therefore, we did not plan it according to the maximal theoretically achievable sterilization effect, but to design a life detection mission to Mars. [8] Wan, L., Wendner, R., & Cusatis, G. A Novel Material for In Situ Construction on Mars: Experiments and Numerical Simulations. 2016, arXiv preprint arXiv:1512.05461	2	1	The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			life6020022_perova	0
+The title has been changed.	2	1	The other potentially inactivating components of the plasma are not specified in the manuscript.			life6020022_perova	0
+In the introduction we already described the problem of the destructiveness of heat sterilization like autoclaving on the organic contents of a soil, which was our motivation to test an alternative method such as low temperature plasma sterilization. D. radiodurans was used as test organism because of its resistance towards UV radiation, which is a part of the sterilizing effect of plasma sterilization. The appropriate passages in the introduction section have been edited.	2	1	Line 38: Deinococcus radiodurans should be written in italic.			life6020022_perova	0
+Since we did not intend to test the influences of the Martian Analog soils on D. radiodurans, but the sterilizing effect of the low temperature plasma treatment on cells intermixed with soil, we did not make a control to determine the effects of the Mars analog soils. The starting values of cell numbers were estimated on samples prepared the same way as the samples for the plasma sterilization (intermixed with soils and desiccated as described in the method section). They were incubated on TYG agar plates like the plasma samples after the treatment. This explanation was missing in the text and has now been added to the appropriate passages in the method section (Chapter 2.2.) for a better understanding.	2	1	The manuscript contains scientific material that can be published as a short communication.			life6020022_perova	0
+Done, thank you.	2	1	Reply: The names are written in italics now, the mistake must have happened during the editing of the text.			life6020022_perova	0
+Yes, we just used one medium (TYG), so we changed it to ‘medium’.	2	1	First of all it is necessary to change the title and at least remove “The testing”.			life6020022_perova	0
+The writing has been changed accordingly.	2	1	Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?			life6020022_perova	0
+Since the bottles containing the Martian Analog soils were autoclaved before use, they should be sterile. We also made a separate test to check for sterility after autoclaving the Martian Analog soils, which has been negative (meaning no contamination could be detected via plating of the autoclaved JSC Mars-1A regolith on TYG medium).	2	1	Reply: The names are written in italics now, the mistake must have happened during the editing of the text.			life6020022_perova	0
+An explanation has been added to the text passage.	2	1	Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?			life6020022_perova	0
+The differences might be caused due to the finer grain size of the other regolith used - particularly P-MRS, which possibly increased the shielding effect. This has been added to the discussion section.	2	1	It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are.			life6020022_perova	0
+I added a description at line 38 – 39.	2	1	From the above, the manuscript then proposes that the first RNA world system may have consisted of two RNA genes of 100 base pairs each (this would probably be better described as genes of 100 nucleotides, unless the author is really proposing the presence of double–stranded RNA?).			life6030029_makarova	0
+I added a description at line 73 – 74.	2	1	The finding that two RNA molecules would have been sufficient to start life, seems a little like the idea that has been proposed of an RNA template and its complementary copy originally functioning as an informational/functional(= ribozymic) pair (see for example: Shay JA, Huynh C, Higgs PG (2015) J Theor Biol.			life6030029_makarova	0
+This comment is in relation to the first comment by reviewer 2. Thus, I added description in lines 99 – 101 and 111 – 112. And I shorten this part to appreciate the comment by reviewer 2.	2	1	There are many quantitative traits affected by many genes.			life6030029_makarova	0
+Actually, the comparison was made for cell-based life-like systems. So, I removed 2 references regarding civilization from this part, and add words at line 243 and 276 – 277. And, I simplify descriptions as shown in line 272.	2	1	Are you saying a cell interacts with the environment but a single gene does not?			life6030029_makarova	0
+This part is important. While it was difficult to simplify, but I tried to simplify section 2.3 as possible and added some descriptions.	2	1	It is only when we get to paragraph 3.1 that we get to the point of the article.			life6030029_makarova	0
+It is not essential that the whole circular network to be incorporated in a life-like system. A part of the network is accelerate by a newly evolved ribozyme is important. I tried to describe this, but it seems not sufficient. So I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 48	2	1	From the origin of life viewpoint, even the simplest of these (the prokaryote) is still very complex.			life6030029_makarova	0
+I added a description at lines 538 – 540.	2	1	I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question.			life6030029_makarova	0
+As I mentioned above, I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 488.	2	1	This links to the introductory section about hydrothermal conditions etc.			life6030029_makarova	0
+Formation of membrane is very important as reviewer 1 mentioned. I added a description at lines 542 – 544, 483 – 488, 544 – 548. line 554 - The case that two genes were essential initially is not made.	2	1	Maybe there is some slightly different nucleic acid-like polymer that is stable in hydrothermal conditions.			life6030029_makarova	0
+Polymerase can only amplify the population of RNA molecules. So, the population of RNA molecules would become of a population consisting of most rapid molecules for replication and then further evolution will struggle. Thus, connection of CCSI with CMIO is essential process to escape from such a non-living state.	2	1	chemistry background (and has published a number of physical chemistry studies investigating the compatibility of the RNA world hypothesis with a hydrothermal vent origin of life), but has also published extensively in the social sciences area, for example applying evolutionary concepts to other 'life-like' systems, such as social insect colonies and human civilizations.			life6030029_makarova	0
+As I agree the reviewer 1, this is important point. And it should be evaluate to identify what process was accelerated at the beginning.	2	1	The conclusion comes back to the point about primitive Earth environment.			life6030029_makarova	0
+I remove some descriptions about hydrothermal environments from Abstract and Conclusions.	2	1	If so, can RNA control the growth and division of the membrane?			life6030029_makarova	0
+This comment is in relation to the thrid comment by reviewer 1. But, reviewer 1 agrees this part. So, I shorten this part.	2	1	There may be a link from one gene to one protein, but one protein is not really a phenotype.			life6030029_makarova	0
+I remove the reference 11 from this part, and moved to line 108. I changed “soft molecules” to “organic molecules”, and changed to “formation and degradation”. The statement was changed to a sentence “… to focus …” shown in line 227.	2	1	 line 73 - the point is made that current RNA World theories do not seem compatible with a hydrothermal origin of life.			life6030029_makarova	0
+8. This is related to the comments about Fig. I added ref 74, 75. In addition, for the size of ribozyme, I added description at line 565 – 566 and ref 67 – 69.	2	1	There are many quantitative traits affected by many genes.			life6030029_makarova	0
+I added a description at line 38 – 39.	2	1	This links to the introductory section about hydrothermal conditions etc.			life6030029_perova	0
+I added a description at line 73 – 74.	2	1	However, I find the manuscript contains a number of weaknesses: One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused.			life6030029_perova	0
+This comment is in relation to the first comment by reviewer 2. Thus, I added description in lines 99 – 101 and 111 – 112. And I shorten this part to appreciate the comment by reviewer 2.	2	1	The idea in caption to Fig 4 that building blocks do not interact directly with the environment seems interesting and plausible, but not very well defined.			life6030029_perova	0
+Actually, the comparison was made for cell-based life-like systems. So, I removed 2 references regarding civilization from this part, and add words at line 243 and 276 – 277. And, I simplify descriptions as shown in line 272.	2	1	However, I find the manuscript contains a number of weaknesses: One of the main issues is that paper suffers from trying to cover too many ideas, and because of this is quite long and unfocused.			life6030029_perova	0
+This part is important. While it was difficult to simplify, but I tried to simplify section 2.3 as possible and added some descriptions.	2	1	I think there is some room for improvement in this diagram.			life6030029_perova	0
+It is not essential that the whole circular network to be incorporated in a life-like system. A part of the network is accelerate by a newly evolved ribozyme is important. I tried to describe this, but it seems not sufficient. So I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 48	2	1	"Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."			life6030029_perova	0
+I added a description at lines 538 – 540.	2	1	Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point.			life6030029_perova	0
+As I mentioned above, I added carefully descriptions at lines 455 – 457, 466 – 471, 483 – 488.	2	1	The input and output would not necessarily need to be controlled by another gene.			life6030029_perova	0
+Formation of membrane is very important as reviewer 1 mentioned. I added a description at lines 542 – 544, 483 – 488, 544 – 548. line 554 - The case that two genes were essential initially is not made.	2	1	Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point.			life6030029_perova	0
+Polymerase can only amplify the population of RNA molecules. So, the population of RNA molecules would become of a population consisting of most rapid molecules for replication and then further evolution will struggle. Thus, connection of CCSI with CMIO is essential process to escape from such a non-living state.	2	1	There are a number of interesting points raised along the way, such as the generally one-to-one correspondence between DNA gene -> mRNA transcript -> protein enzyme -> single reaction (although many enzymes are often involved in a single pathway which transforms/produces a single substrate/product).			life6030029_perova	0
+As I agree the reviewer 1, this is important point. And it should be evaluate to identify what process was accelerated at the beginning.	2	1	In summary, I think there are a lot of interesting points here, but the article could use some streamlining to emphasize the key new ideas.			life6030029_perova	0
+I remove some descriptions about hydrothermal environments from Abstract and Conclusions.	2	1	Fig 8 is very reminiscent of Fig 1 of Wu and Higgs (2009) J Mol Evol 69:541-554 and Fig 1 of Wu and Higgs (2011) Astrobiology 11:895-906.			life6030029_perova	0
+This comment is in relation to the thrid comment by reviewer 1. But, reviewer 1 agrees this part. So, I shorten this part.	2	1	There are a number of interesting points raised along the way, such as the generally one-to-one correspondence between DNA gene -> mRNA transcript -> protein enzyme -> single reaction (although many enzymes are often involved in a single pathway which transforms/produces a single substrate/product).			life6030029_perova	0
+I remove the reference 11 from this part, and moved to line 108. I changed “soft molecules” to “organic molecules”, and changed to “formation and degradation”. The statement was changed to a sentence “… to focus …” shown in line 227.	2	1	This section seems to mix up two important questions.			life6030029_perova	0
+8. This is related to the comments about Fig. I added ref 74, 75. In addition, for the size of ribozyme, I added description at line 565 – 566 and ref 67 – 69.	2	1	The input and output would not necessarily need to be controlled by another gene.			life6030029_perova	0
+To the best of our knowledge, rare earth doped yttrium iron garnet is still the main material used for magneto-optical devices. They include bismuth doped YIG, bismuth, terbium doped YIG and cerium doped YIG. Other materials such as Wely semimatels are still under theoretical study[1]. We have added comments to these materials in the manuscript. Revisions: Page 1, line 21, added “At present, rare earth doped yttrium iron garnet (RIG) is the most widely used magneto-optical material in integrated MO devices.” Point 2:	2	1	By replacing Y3+ ions with Dy3+ ions in Ce:YIG, authors demonstrate less than ±5% variation of the thin film Faraday rotation from the temperature of 25 ℃ to 70 ℃, compared to ~20% in Ce:YIG.			ma15051691_makarova	0
+Absoultly there is also temperature dependence of the Faraday rotation when shift to other wavelengths. However, the transparency window of this material is in the 1550 nm wavelength range. When moving to shorter wavelengths, the absorption of this material increases sharply, making them less practical for photonic device applications.	2	1	demonstrate a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications.			ma15051691_makarova	0
+previously reported that the compensation temperature of DyIG is Tcomp = 225 K [2,3], which could decrease the Faraday rotation angle of Dy:CeIG in this work at the temperature below 300 K. Therefore, a higher Faraday rotation may be related to the increase of the saturation magnetization of this material in this temperature range. Revisions:Page 5, line 170-173, added “The increase of the Faraday rotation below 40 ℃ is possibly due to the increase of the magnetization of this material at this temperature range, considering a compensation temperature of 225 K in Dy3Fe5O12 [19,20].”. References: Okamura, Y.; Minami, S.; Kato, Y.; Fujishiro, Y.; Kaneko, Y.; Ikeda, J.; Muramoto, J.; Kaneko, R.; Ueda, K.; Kocsis, V.; et al. Giant magneto-optical responses in magnetic Weyl semimetal Co3Sn2S2. Nat Commun 2020, 11, 4619, doi:10.1038/s41467-020-18470-0. Sayetat, F. Huge magnetostriction in Tb3Fe5O12, Dy3Fe5O12, Ho3Fe5O12, Er3Fe5O12 garnets. Journal of magnetism and magnetic materials 1986, 58, 334-346.	2	1	This manuscript relates to the effect of dysprosium substitution in Ce:YIG thin films and in particular to the temperature dependance of their Faraday rotation.			ma15051691_makarova	0
+We have updated the references and revised this in the context. Revisions: Page 1, line 21, revised to “for silicon integrated photonic circuits (PICs) [1-3].”. Page 1, line 24, revised to “including optical isolators [4-6]”. Page 1, line 41, revised to “which results in reduced bandwidth and isolation ratio [13].”. Page 1, line 45, revised to “in a temperature range of 20-60 ℃ [10].”. Page 5, line 148, revised to “measure the temperature stability of Dy:CeIG and Ce:YIG films[11].”.	2	1	"Author Response Reviewer #1 (Comments to the Author): The article ""Dysprosium substituted Ce:YIG thin films for temperature in-2 sensitive integrated optical isolator applications contains the results"" presents a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications."			ma15051691_makarova	0
+We have revised this in the context. Revisions: Page 3, line 98, revised to “where is the Faraday rotation angle of MO films at room temperature. is the NRPS at room temperature.” Point 3:	2	1	Ostorero, J.; Escorne, M.; Pecheron‐Guegan, A.; Soulette, F.; Le Gall, H. Dy3Fe5O12 garnet thin films grown from sputtering of metallic targets.			ma15051691_makarova	0
+Thanks for correcting this. “X-ray diffraction spectra” has been revised into “X-ray diffraction patterns”. Revisions: Page 3, line 109, revised to “X-ray diffraction patterns”.	2	1	"Author Response Reviewer #1 (Comments to the Author): The article ""Dysprosium substituted Ce:YIG thin films for temperature in-2 sensitive integrated optical isolator applications contains the results"" presents a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications."			ma15051691_makarova	0
+Thanks for the commerts. The saturation magnetization of rare-earth doped YIG affects the Faraday rotation angle. Sayetat et al. and Ostorero et al. previously reported that the compensation temperature of DyIG is Tcomp = 225 K [1,2], which could decrease the Faraday rotation angle of Dy:CeIG in this work at the temperature below 300 K. Therefore, a higher Faraday rotation may be related to the increase of the saturation magnetization of this material in this temperature range. Revisions: Page 5, line 170-173, added “The increase of the Faraday rotation below 40 ℃ is possibly due to the increase of the magnetization of this material at this temperature range, considering a compensation temperature of 225 K in Dy3Fe5O12 [19,20].”.	2	1	It is in general well written and the work deserves publication.			ma15051691_makarova	0
+Ostorero, J.; Escorne, M.; Pecheron‐Guegan, A.; Soulette, F.; Le Gall, H. Dy3Fe5O12 garnet thin films grown from sputtering of metallic targets. Journal of Applied Physics 1994, 75, 6103-6105. Thanks for pointing this out. We have revised the Figure 3. Revisions: Page 6, revised the horizontal scale of Figure 3 (c) and updated Figure 3 (c). References: Sayetat, F. Huge magnetostriction in Tb3Fe5O12, Dy3Fe5O12, Ho3Fe5O12, Er3Fe5O12 Journal of magnetism and magnetic materials 1986, 58, 334-346.	2	1	We answer the reviewer’s questions as follows.			ma15051691_makarova	0
+Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS) obey the same Bragg's Law (2dsinθ=nλ), but with different distance from sample to detector. With a distance about 384 mm in this work, the WAXS can measure the structure from 0.12 nm to 0.45 nm, which correspond to the crystal lattice, while SAXS can measure the structure from 10 nm to 80 nm, with a distance of 2484 mm. In a SAXS curve, the strength reflect the number of defects, while the q range reflect the size of the defects.	2	1	), we described in detail how to calculate the specific surface area and volume fraction.			ma15124258_makarova	0
+Thanks for your question, we looked up several literature and couldn't find a fixed usage of cracks and voids, and in this article, we think that using pores would completely cover our research objectives, so we deleted the cracks in this paper. Computed microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	Do you have another proof for your conclusions.			ma15124258_makarova	0
+Yes, the pores are assumed to be spherical in the fitting of SAXS data, because the spherical model is the simplest and widely used model in the SAXS fitting. The voids are cylindrical that is close to a spherical, so we can get a relatively accurate size distribution by assuming to spherical.	2	1	The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			ma15124258_makarova	0
+Yes, we had an experiment of differential scanning calorimetry (DSC) to characterize the process, as shown in Figure 1. According to the result from DSC, there were an endothermic peak of phase transformation between the temperature range of 151~172 ℃, which are not exactly consistent with the results from in this paper. We think the difference in temperature is result from the different sample amount and heating rate. As with X-ray diffraction (XRD), the WAXS can be used to characterize the content of different phase in the phase transformation. So, we think the results in our paper is reasonable.	2	1	Could you use X-ray microtomography on your samples?			ma15124258_makarova	0
+SAXS can be used to measure the defect size and number of defects with rational theory and adequate validation in many	2	1	Some changes in propagation of pores caused by both the phase transitions and temperature were observed by an appropriate experimental technique.			ma15124258_makarova	0
+The colors in figure 1 and 3 represent the strength of scattering, while the colors in other figures just represent different date.	2	1	As you can tell by these questions, my main issue is related to the specific form of the thermal damage.			ma15124258_makarova	0
+The result of SAXS is a reflection of two-phase with different electron density. In our paper, the CL-20 powders are immersed in GPL107 with a approximately equal electron density with CL-20 crystals, so the SAXS reflects the nano-scale pores inside CL-20 powders. Thereby, the increase of strength in SAXS curve reflects the increase in number of internal nano-scale pores. According to the principle of SAXS, the scale of defects is correlated with the q range, so the growth into larger defects will appear in other q range. X-ray microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	It indeed the case, but I could not find any discussion on this issue.			ma15124258_makarova	0
+The GPL107 is resistant to high temperatures about 400 ℃, and we used the SAXS patterns of GPL107 at different temperature as the background to correct any bias at different temperature.	2	1	199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.			ma15124258_makarova	0
+According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity in lager q range can be explained into the increase in more small voids. And also, our fitting result can provide more obvious display.	2	1	Thus, I can recommend it for publication after a minor revision.			ma15124258_makarova	0
+In our paper, spherical model was just a choice in the SAXS date fitting, so we did not provide more illustrate, but a reference (L A Feigin and D I Svergun, Structure Analysis by Small-Angle X-Ray and Neutron Scattering, Plenum Press, New York, 1987) was provide in the manuscript. On line 173, we gave a conclusion “the number of pores increased, but the size distribution did not change.” We did not consider the pore volume, but we can deduce that the pores volume increase as the increase of the number of pores.	2	1	Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?			ma15124258_makarova	0
+Thanks again for your attention, According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity without the change of q range can be explained into the number of pores increasing, while the existing pores increasing in volume would result into an change of q to low range.	2	1	Review of the manuscript Investigation on the Evolution of Nano-scale Defects of CL-20 Crystals under Thermal Treatment by Wide/Small-Angle X-ray Scattering, Haobin Zhang, Hongfan Wang … The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			ma15124258_makarova	0
+Thanks for your comments, we added three reference about recent work published in 2021 and 2022.	2	1	Could computed microtomography be used to confirm some of the experimental results?			ma15124258_makarova	0
+Thanks for you comment, I have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety after thermal treatment in the conclusion.	2	1	199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.”From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.			ma15124258_makarova	0
+Thanks for you comments, and I have added the particle size in the manuscript.	2	1	Is the temperature high enough to cause decomposition effects?			ma15124258_makarova	0
+The electron density of CL-20 and GPL107 are calculated according to the classical electron radius ( with a size of 2.818 E-13 cm) and the electron number per volume, the detailed information can refer to our previous work (Molecules 2020, 25 , 443.)	2	1	Every available suggestion has been answered carefully and corrected in the article.			ma15124258_makarova	0
+In our previous work (Molecules 2020, 25, 443. ), we described in detail how to calculate the specific surface area and volume fraction. In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.	2	1	Some changes in propagation of pores caused by both the phase transitions and temperature were observed by an appropriate experimental technique.			ma15124258_makarova	0
+“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20. To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.” We have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after thermal treatment in the conclusion.	2	1	Could you use X-ray microtomography on your samples?			ma15124258_makarova	0
+Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS) obey the same Bragg's Law (2dsinθ=nλ), but with different distance from sample to detector. With a distance about 384 mm in this work, the WAXS can measure the structure from 0.12 nm to 0.45 nm, which correspond to the crystal lattice, while SAXS can measure the structure from 10 nm to 80 nm, with a distance of 2484 mm. In a SAXS curve, the strength reflect the number of defects, while the q range reflect the size of the defects.	2	1	Response to Reviewer' Comments Dear Reviewer, We sincerely appreciate your comments and suggestions on the manuscript!			ma15124258_perova	0
+Thanks for your question, we looked up several literature and couldn't find a fixed usage of cracks and voids, and in this article, we think that using pores would completely cover our research objectives, so we deleted the cracks in this paper. Computed microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	Response: In our previous work (Molecules 2020, 25, 443.			ma15124258_perova	0
+Yes, the pores are assumed to be spherical in the fitting of SAXS data, because the spherical model is the simplest and widely used model in the SAXS fitting. The voids are cylindrical that is close to a spherical, so we can get a relatively accurate size distribution by assuming to spherical.	2	1	“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20.			ma15124258_perova	0
+Yes, we had an experiment of differential scanning calorimetry (DSC) to characterize the process, as shown in Figure 1. According to the result from DSC, there were an endothermic peak of phase transformation between the temperature range of 151~172 ℃, which are not exactly consistent with the results from in this paper. We think the difference in temperature is result from the different sample amount and heating rate. As with X-ray diffraction (XRD), the WAXS can be used to characterize the content of different phase in the phase transformation. So, we think the results in our paper is reasonable.	2	1	Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			ma15124258_perova	0
+SAXS can be used to measure the defect size and number of defects with rational theory and adequate validation in many field.	2	1	Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			ma15124258_perova	0
+The colors in figure 1 and 3 represent the strength of scattering, while the colors in other figures just represent different date.	2	1	It indeed the case, but I could not find any discussion on this issue.			ma15124258_perova	0
+The result of SAXS is a reflection of two-phase with different electron density. In our paper, the CL-20 powders are immersed in GPL107 with a approximately equal electron density with CL-20 crystals, so the SAXS reflects the nano-scale pores inside CL-20 powders. Thereby, the increase of strength in SAXS curve reflects the increase in number of internal nano-scale pores. According to the principle of SAXS, the scale of defects is correlated with the q range, so the growth into larger defects will appear in other q range. X-ray microtomography is usually used to confirm the voids in micron-scale but this paper focuses on nano-scale voids.	2	1	The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			ma15124258_perova	0
+The GPL107 is resistant to high temperatures about 400 ℃, and we used the SAXS patterns of GPL107 at different temperature as the background to correct any bias at different temperature.	2	1	I recommend publishing the manuscript after minor the revision.			ma15124258_perova	0
+According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity in lager q range can be explained into the increase in more small voids. And also, our fitting result can provide more obvious display.	2	1	How do you results compare to phase transformation measured using DSC?			ma15124258_perova	0
+In our paper, spherical model was just a choice in the SAXS date fitting, so we did not provide more illustrate, but a reference (L A Feigin and D I Svergun, Structure Analysis by Small-Angle X-Ray and Neutron Scattering, Plenum Press, New York, 1987) was provide in the manuscript. On line 173, we gave a conclusion “the number of pores increased, but the size distribution did not change.” We did not consider the pore volume, but we can deduce that the pores volume increase as the increase of the number of pores.	2	1	), we described in detail how to calculate the specific surface area and volume fraction.			ma15124258_perova	0
+Thanks again for your attention, According to the principle of SAXS, the scale of defects is correlated with the q range, so the increase in scattering intensity without the change of q range can be explained into the number of pores increasing, while the existing pores increasing in volume would result into an change of q to low range.	2	1	How do you results compare to phase transformation measured using DSC?			ma15124258_perova	0
+Thanks for your comments, we added three reference about recent work published in 2021 and 2022.	2	1	Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			ma15124258_perova	0
+Thanks for you comment, I have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety after thermal treatment in the conclusion.	2	1	To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”			ma15124258_perova	0
+Thanks for you comments, and I have added the particle size in the manuscript.	2	1	Review of the manuscript Investigation on the Evolution of Nano-scale Defects of CL-20 Crystals under Thermal Treatment by Wide/Small-Angle X-ray Scattering, Haobin Zhang, Hongfan Wang … The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			ma15124258_perova	0
+The electron density of CL-20 and GPL107 are calculated according to the classical electron radius ( with a size of 2.818 E-13 cm) and the electron number per volume, the detailed information can refer to our previous work (Molecules 2020, 25 , 443.)	2	1	Every available suggestion has been answered carefully and corrected in the article.			ma15124258_perova	0
+In our previous work (Molecules 2020, 25, 443. ), we described in detail how to calculate the specific surface area and volume fraction. In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.	2	1	Every available suggestion has been answered carefully and corrected in the article.			ma15124258_perova	0
+We have supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after thermal treatment in the conclusion. “During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20. To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”	2	1	Do you have another proof for your conclusions.			ma15124258_perova	0
+There was a question related to electronic devices (eCigarettes) but we missed to add this data. Only 0.5% of participants used eCigaretes or similar devices. We added this sentence in the section Results.	2	1	Please provide a conclusion based on obtained findings and please avoid overwhelming conclusions Response 8.			medicina58040502_makarova	0
+It has been showed that higher exposure to secondhand smoking increased the risk of smoking. The multivariable logistic regression analysis showed that variables such as secondhand smoke at home, secondhand smoke at faculty, and secondhand smoke at public spaces can determine whether students smoke or not. It is also obvious that non-smokers avoid being in closed smoking places, but this was not subject of this study.	2	1	This is contrary to accepted public health policy internationally.			medicina58040502_makarova	0
+Your comment is very logical. Cigarette smoking, sedentary life-style and obesity are the major public health concerns, particularly in the Balkan region. This was a reason that we have provided some additional comments and recommendations for further actions at the very end of the Discussion section.	2	1	Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals) Response 6.			medicina58040502_makarova	0
+The “Aim of the study” has been changed completely.	2	1	This is a well-designed and executed survey of university cigarette smoking in a country with above-average poverty.			medicina58040502_makarova	0
+To reply on this comment and to provide readers with more precise data we made some changes in two sections. In the subsection “Sample size” of the section “Materials and methods” we emphasized that “students from 16 faculties of the University of Banja Luka were participants in our study”. In section “Results” we have added a new sentence that “Majority of study participants were medical students (41.4%) and the rest were students from other faculties (58.6%).” Comment 3.	2	1	Medicina 2021, 57, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Lastname Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.			medicina58040502_makarova	0
+This has been already presented in Table 2. and explained by asterisks (**) below the table: * Students who smoked at least one day in 30 days prior to the survey. ** Students who smoke every day and desired to smoke always or sometimes upon waking up, and where smoking time after waking up was within one day. If necessary the English version of the Questionnaire could be provided as Supplementary material.	2	1	In Table 5 please provide thr number of subjects (n=.....) Response 5.			medicina58040502_makarova	0
+We have made some changes in the section “Results” to avoid data duplication.	2	1	In Table 5 please provide thr number of subjects (n=.....) Response 5.			medicina58040502_makarova	0
+The number of subjects (N) has been added.	2	1	Response to Reviewer 2 I would like to thank Reviewer 2 for very detailed and useful comments which will definitely improve our manuscript.			medicina58040502_makarova	0
+The contents of Table 6 and Table 7 have been rearranged and comprised into one table which is now Table 6.	2	1	Is the omission of any items referring to experience/use of eCigarette/vaping.			medicina58040502_makarova	0
+In the last paragraph of the section “Discussion” we have proposed some practical implications and recommendations for further actions.	2	1	In Table 5 please provide thr number of subjects (n=.....) Response 5.			medicina58040502_makarova	0
+The section “Conclusion” has been completely revised and rearranged according to the reviewer’s comment.	2	1	Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous.			medicina58040502_makarova	0
+There was a question related to electronic devices (eCigarettes) but we missed to add this data. Only 0.5% of participants used eCigaretes or similar devices. We added this sentence in the section Results.	2	1	Is the omission of any items referring to experience/use of eCigarette/vaping.			medicina58040502_perova	0
+It has been showed that higher exposure to secondhand smoking increased the risk of smoking. The multivariable logistic regression analysis showed that variables such as secondhand smoke at home, secondhand smoke at faculty, and secondhand smoke at public spaces can determine whether students smoke or not. It is also obvious that non-smokers avoid being in closed smoking places, but this was not subject of this study.	2	1	Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.			medicina58040502_perova	0
+Your comment is very logical. Cigarette smoking, sedentary life-style and obesity are the major public health concerns, particularly in the Balkan region. This was a reason that we have provided some additional comments and recommendations for further actions at the very end of the Discussion section.	2	1	Please provide the most important results in the text, and the rest of them are present in Tables.			medicina58040502_perova	0
+The “Aim of the study” has been changed completely.	2	1	Results were consistent with previous studies of similar target populations.			medicina58040502_perova	0
+To reply on this comment and to provide readers with more precise data we made some changes in two sections. In the subsection “Sample size” of the section “Materials and methods” we emphasized that “students from 16 faculties of the University of Banja Luka were participants in our study”. In section “Results” we have added a new sentence that “Majority of study participants were medical students (41.4%) and the rest were students from other faculties (58.6%).” Comment 3.	2	1	Response to Reviewer 1 I would like to thank Reviewer 1 for very detailed and useful comments which will definitely improve our manuscript.			medicina58040502_perova	0
+Response 3. This has been already presented in Table 2. and explained by asterisks (**) below the table: * Students who smoked at least one day in 30 days prior to the survey. ** Students who smoke every day and desired to smoke always or sometimes upon waking up, and where smoking time after waking up was within one day. If necessary the English version of the Questionnaire could be provided as Supplementary material.	2	1	Response to Reviewer 1 I would like to thank Reviewer 1 for very detailed and useful comments which will definitely improve our manuscript.			medicina58040502_perova	0
+We have made some changes in the section “Results” to avoid data duplication.	2	1	Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous.			medicina58040502_perova	0
+The number of subjects (N) has been added.	2	1	Is the omission of any items referring to experience/use of eCigarette/vaping.			medicina58040502_perova	0
+The contents of Table 6 and Table 7 have been rearranged and comprised into one table which is now Table 6.	2	1	Results were consistent with previous studies of similar target populations.			medicina58040502_perova	0
+In the last paragraph of the section “Discussion” we have proposed some practical implications and recommendations for further actions.	2	1	This is the opposite of health promotion one expects from all healthcare providers - especially physicians.			medicina58040502_perova	0
+The section “Conclusion” has been completely revised and rearranged according to the reviewer’s comment.	2	1	Recommendations did not go beyond the data and human subjects protections were adequate.			medicina58040502_perova	0
+thank you for reviewer’s comments, we made changes accordingly. We added statistical analysis for CNA association between case and reference groups (please see page 5).	2	1	The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			medsci4030012_makarova	0
+For the questions of “is the deletion burden observed really exceptional in a population sample of similar ancestry”, we agree with the reviewer’s comment. We did the analysis a couple of years ago, at that time, there is no CNV data from the 1000 genome for Mexicans, Colombians, Puerto Ricans and Peruvians, however, there is data for the admixed populations, thus we used this for this study. There might be some difficulty to do more analysis since our statistician and bioinformatics person left the department and we are in the process of hiring somebody for this role. At the same, we continue in recruiting more subjects with cervical cancers to further confirm our findings. Moreover, in the discussion, we have list this as one of the limitations in page 8 “2) there might be bias of the CNA identified in cervical cancer for the Mexican Americans since we used the 1000 Genome admixed populations, not Mexican Americans, thus, we current recruit more subjects with cervical cancer from the same population and plan to validate the findings in more samples;” 2)	2	1	It is not clear which statistical method is used for the significant results in Table 2.			medsci4030012_makarova	0
+For the question of “is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies”, our response is that we did search of CNAs observed in other cancer based on TCGA studies, although there are limited studies, we addressed this issue in the discussion section (page 8) “…Using the TCGA data, a recent study identified nine regions of deletion that were unique to ER+ post menopause tumors in patients with breast cancer, including deletion in 7p22.3 where our newly identified deletion in cases only located and it contains known tumor suppressor gene”.	2	1	I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			medsci4030012_makarova	0
+great suggestions, thank the reviewer. Now we added a sentence of “Moreover, the pathway analysis revealed endometrial cancer and estrogen signaling pathways associated with this cancer (P < 0.05) using the KEGG” in the abstract as the reviewer’s suggestion. In addition, in the introduction section, we added “We are aware of the limited number of cases and lack of control group. Thus future a large study with a control sample and more cases as methodological alternative is needed”. (Please see page 3) Minor comments: 1-	2	1	Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			medsci4030012_makarova	0
+as the reviewer’s suggestion, we changed “Latino” to “Mexican American” in the text of the manuscript and table	2	1	7) Lines 238-242 are replicated in discussion lines 376-380.			medsci4030012_makarova	0
+now we change “Latino population” to “Mexican American” as the reviewer’s suggestion	2	1	The authors need to define “pre-cancer” in materials.			medsci4030012_makarova	0
+thank you for the comments. We also realized this issue which is due to that the color green overlaps with the other colors, thus it cannot be distinguished, and we have tried different color combinations without help. Now we made changes of the figure and legend.	2	1	My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			medsci4030012_makarova	0
+thanks, we removed the % sign in table cells in Table 2b	2	1	You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.			medsci4030012_makarova	0
+thank you, reviewer, this is a typo, we corrected it.	2	1	It is not clear if there is one or two groups but just one percentage.			medsci4030012_makarova	0
+yes, we added up to six key words now	2	1	However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.			medsci4030012_makarova	0
+we made changes based on the reviewer’s suggestion, please see statistical method in page 5 and result in page 6	2	1	The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry.			medsci4030012_makarova	0
+thanks, we provided definition for pre-cancer in materials, which include patients with CIN I, II, and III.	2	1	I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			medsci4030012_makarova	0
+thanks, we added “…using statistical analyses described in the method section”	2	1	It is not clear which statistical method is used for the significant results.			medsci4030012_makarova	0
+thanks, the reviewer. Now we removed redundancy statement.	2	1	The PCA they show is probably capturing those differences instead of real differences given by disease status and, by the way, I question the usefulness of this PCA.			medsci4030012_makarova	0
+thank you for reviewer’s comments, we made changes accordingly. We added statistical analysis for CNA association between case and reference groups (please see page 5).	2	1	My suggestion is to offer a better background for these results by trying to answer two main questions: 1) is the deletion burden observed really exceptional in a population sample of similar ancestry?			medsci4030012_perova	0
+For the questions of “is the deletion burden observed really exceptional in a population sample of similar ancestry”, we agree with the reviewer’s comment. We did the analysis a couple of years ago, at that time, there is no CNV data from the 1000 genome for Mexicans, Colombians, Puerto Ricans and Peruvians, however, there is data for the admixed populations, thus we used this for this study. There might be some difficulty to do more analysis since our statistician and bioinformatics person left the department and we are in the process of hiring somebody for this role. At the same, we continue in recruiting more subjects with cervical cancers to further confirm our findings. Moreover, in the discussion, we have list this as one of the limitations in page 8 “2) there might be bias of the CNA identified in cervical cancer for the Mexican Americans since we used the 1000 Genome admixed populations, not Mexican Americans, thus, we current recruit more subjects with cervical cancer from the same population and plan to validate the findings in more samples;” 2)	2	1	Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			medsci4030012_perova	0
+For the question of “is the deletion burden observed similar to those for other cancers? Then again there must be estimates of deletion burden for different types of cancers from TCGA studies”, our response is that we did search of CNAs observed in other cancer based on TCGA studies, although there are limited studies, we addressed this issue in the discussion section (page 8) “…Using the TCGA data, a recent study identified nine regions of deletion that were unique to ER+ post menopause tumors in patients with breast cancer, including deletion in 7p22.3 where our newly identified deletion in cases only located and it contains known tumor suppressor gene”.	2	1	The experimental design is adequate and the statistical methods are updated.			medsci4030012_perova	0
+Now we added a sentence of “Moreover, the pathway analysis revealed endometrial cancer and estrogen signaling pathways associated with this cancer (P < 0.05) using the KEGG” in the abstract as the reviewer’s suggestion. In addition, in the introduction section, we added “We are aware of the limited number of cases and lack of control group. Thus future a large study with a control sample and more cases as methodological alternative is needed”. (Please see page 3) Minor comments: 1-	2	1	The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			medsci4030012_perova	0
+as the reviewer’s suggestion, we changed “Latino” to “Mexican American” in the text of the manuscript and table	2	1	However, I think that the manuscript needs some substantial revisions before it can be published.			medsci4030012_perova	0
+now we change “Latino population” to “Mexican American” as the reviewer’s suggestion	2	1	They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			medsci4030012_perova	0
+thank you for the comments. We also realized this issue which is due to that the color green overlaps with the other colors, thus it cannot be distinguished, and we have tried different color combinations without help. Now we made changes of the figure and legend.	2	1	However, I think that the manuscript needs some substantial revisions before it can be published.			medsci4030012_perova	0
+thanks, we removed the % sign in table cells in Table 2b	2	1	The authors need to define “pre-cancer” in materials.			medsci4030012_perova	0
+thank you, reviewer, this is a typo, we corrected it.	2	1	They found that the burden of deletions in these patients is much higher than expected, as compared with CNV burden in HapMap populations.			medsci4030012_perova	0
+yes, we added up to six key words now	2	1	Then again there must be estimates of deletion burden for different types of cancers from TCGA studies.			medsci4030012_perova	0
+we made changes based on the reviewer’s suggestion, please see statistical method in page 5 and result in page 6	2	1	The authors need to give more details about how to use PCA in statistical methods.			medsci4030012_perova	0
+thanks, we provided definition for pre-cancer in materials, which include patients with CIN I, II, and III.	2	1	The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			medsci4030012_perova	0
+thanks, we added “…using statistical analyses described in the method section”	2	1	2- The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract.			medsci4030012_perova	0
+thanks, the reviewer. Now we removed redundancy statement.	2	1	Tobari and colleagues studied the CNV burden in patients with cervical cancer from Mexican American ancestry.			medsci4030012_perova	0
+We have attempted to improve the language and hope we have found all significant errors.	2	1	Correct subscripts and superscripts in the experimental also check the spacings to be consistent.			molecules26247651_makarova	0
+The characterization of the compounds is provided together with the NMR spectra as Suppl. Mat. S21.	2	1	Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.			molecules26247651_makarova	0
+Spectra have been included in Suppl. Mat. S21.	2	1	Full characterization of all synthesized compounds is required in the experimental section.			molecules26247651_makarova	0
+A new Scheme 1 has been produced with the recommended format. The molecular structures in Figures 1 and 3 have been modified accordingly.	2	1	Check the font of the reference numbers in the text and correct			molecules26247651_makarova	0
+The misalignment is a result of the editorial procedure. We have tried to improve the alignment.	2	1	The rationale for undertaking the research is a bit puzzling.			molecules26247651_makarova	0
+Done.	2	1	All the 1HNMR and 13CNMR must be attached as supplementary information.			molecules26247651_makarova	0
+We appreciate the suggestion. However, we do not think we have time to perform a proper analysis of XRD data, considering solid state effects, crystal packing, etc.	2	1	Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.			molecules26247651_makarova	0
+In the submitted MSWord document the font was Times New Roman for text and reference numbers. Something has happened during the editorial procedure when changing to Palatino Linotype. Not all reference numbers were changed correctly. We have tried to find and correct the errors.	2	1	"Minor: 183: should be SD = 34.3 cm-1186: """"are reasonable"""": Taking into account that (i) this fit is rather unphysical(ii) moving the points by say 50cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."""			molecules26247651_makarova	0
+This concerns the experimental section, S21, in the Supplementary Material document. We have scrutinized the text and corrected the errors. We have also corrected two typos in the main manuscript.	4	1	The cell values do not align please correct them.			molecules26247651_makarova	0
+The background for the investigation is described in the Introduction section. As indicated by the Reviewer, the present study is an extension of our previous OH···O binding studies. We have for some time wished to investigate the corresponding NH···O systems. The NH···O type of linkage has the advantage that a wide range of compounds can be investigated; in general, systems of the type NH···O=C with an intervening double bond are not tautomeric, in contrast to the corresponding OH···O=C systems. We have added a remark on this aspect to the Introduction section. - The ‘delay’ of 10 years is due to technical and personal factors of no relevance to the reported results.	2	1	Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			molecules26247651_makarova	0
+"We are aware of the controversial nature of this result. We also expected a linear relationship similar to the one observed by Dudek. However, we have in this work extended the range of compounds to include species of types J, K, and L, apparently resulting in the observed non-linear relationship. The correlation in Fig. 2 involves only experimental values, most of them from the literature, and we have no reason to doubt their reliability. The exponential regression yields R2 = 0.975, SD = 34.3 cm–1, while the corresponding linear regression gives R2 = 0.837, SD = 93.3 cm–1. Hence, the indication of a non-linear correlation is evident. We have added the sentence ""Probably the extension of the range of compounds to include species of the types J, K, and L has led to the non-linear correlation” to the first paragraph on page 13."	2	1	The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			molecules26247651_makarova	0
+"Our attention was drawn to Ref. [54] where Grimme et al. state that ""The [DFT-D3] method has been assessed on standard benchmark sets for inter- and intramolecular noncovalent interactions"". We thus found it relevant to try out B3LYP-D3 for the present study of ""intramolecular noncovalent interactions"". But the Reviewer finds that these calculations ""make no sense here"", so we have removed these results from the manuscript. The Reviewer suggests that we check the functional B3PW91 (in the Introduction we actually mention B3PW91 as one of the two functionals recommended by Scott and Radom). We have performed additional B3PW91/6-311++G(d,p) calculations on a series of seven compounds covering a wide range of NH stretching wavenumbers. The computed harmonic wavenumbers are essentially linearly related to those obtained with B3LYP (R2 = 0.9999, SD = 2.6 cm–1). We have added a remark on this result to the last paragraph of Section 3.2."	2	1	In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			molecules26247651_makarova	0
+Corrected!	2	1	The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			molecules26247651_makarova	0
+"The sentence with ""are reasonable"" has been removed."	2	1	Use 4 significant figures for the RNH values in table 2 7.			molecules26247651_makarova	0
+The orientation of the axes was chosen according to the trend reported by Novak [56]: Decreasing NH stretching wavenumber corresponds to decreasing isotope ratio.	2	1	Here, calculations are performed for isolated molecules.			molecules26247651_makarova	0
+We have attempted to improve the language and hope we have found all significant errors.	2	1	Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major:			molecules26247651_perova	0
+The characterization of the compounds is provided together with the NMR spectra as Suppl. Mat. S21.	2	1	There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.2.			molecules26247651_perova	0
+Spectra have been included in Suppl.	2	1	I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			molecules26247651_perova	0
+A new Scheme 1 has been produced with the recommended format. The molecular structures in Figures 1 and 3 have been modified accordingly.	2	1	Full characterization of all synthesized compounds is required in the experimental section.			molecules26247651_perova	0
+The misalignment is a result of the editorial procedure. We have tried to improve the alignment.	2	1	Correct subscripts and superscripts in the experimental also check the spacings to be consistent.			molecules26247651_perova	0
+Done.	2	1	As indicated by the Reviewer, the present study is an extension of our previous OH···O binding studies.			molecules26247651_perova	0
+We appreciate the suggestion. However, we do not think we have time to perform a proper analysis of XRD data, considering solid state effects, crystal packing, etc.	2	1	There are some questions about the manuscript as follows:1.			molecules26247651_perova	0
+In the submitted MSWord document the font was Times New Roman for text and reference numbers. Something has happened during the editorial procedure when changing to Palatino Linotype. Not all reference numbers were changed correctly. We have tried to find and correct the errors.	2	1	It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.			molecules26247651_perova	0
+This concerns the experimental section, S21, in the Supplementary Material document. We have scrutinized the text and corrected the errors. We have also corrected two typos in the main manuscript.	4	1	In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			molecules26247651_perova	0
+"We are aware of the controversial nature of this result. We also expected a linear relationship similar to the one observed by Dudek. However, we have in this work extended the range of compounds to include species of types J, K, and L, apparently resulting in the observed non-linear relationship. The correlation in Fig. 2 involves only experimental values, most of them from the literature, and we have no reason to doubt their reliability. The exponential regression yields R2 = 0.975, SD = 34.3 cm–1, while the corresponding linear regression gives R2 = 0.837, SD = 93.3 cm–1. Hence, the indication of a non-linear correlation is evident. We have added the sentence ""Probably the extension of the range of compounds to include species of the types J, K, and L has led to the non-linear correlation” to the first paragraph on page 13."	2	1	In this article, the authors describe the spectral characteristics of NH in the hydrogen bridge NH...O.			molecules26247651_perova	0
+"Our attention was drawn to Ref. [54] where Grimme et al. state that ""The [DFT-D3] method has been assessed on standard benchmark sets for inter- and intramolecular noncovalent interactions"". We thus found it relevant to try out B3LYP-D3 for the present study of ""intramolecular noncovalent interactions"". But the Reviewer finds that these calculations ""make no sense here"", so we have removed these results from the manuscript. The Reviewer suggests that we check the functional B3PW91 (in the Introduction we actually mention B3PW91 as one of the two functionals recommended by Scott and Radom). We have performed additional B3PW91/6-311++G(d,p) calculations on a series of seven compounds covering a wide range of NH stretching wavenumbers. The computed harmonic wavenumbers are essentially linearly related to those obtained with B3LYP (R2 = 0.9999, SD = 2.6 cm–1). We have added a remark on this result to the last paragraph of Section 3.2."	2	1	3. All the 1HNMR and 13CNMR must be attached as supplementary information.			molecules26247651_perova	0
+Corrected!	2	1	Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.			molecules26247651_perova	0
+"The sentence with ""are reasonable"" has been removed."	2	1	The rationale for undertaking the research is a bit puzzling.			molecules26247651_perova	0
+The orientation of the axes was chosen according to the trend reported by Novak [56]: Decreasing NH stretching wavenumber corresponds to decreasing isotope ratio.	2	1	We have for some time wished to investigate the corresponding NH···O systems.			molecules26247651_perova	0
+We appreciate the recommendation and comments. We corrected the above-mentioned correction as suggested by the reviewer and presented as follows, 1H NMR (400 MHz, DMSO-d6, ppm): δ 11.43 (s, 2H), 8.34 (m, 2H), 8.28 (m, 1H), 7.65 (d, 2H, J = 4.8 Hz), 7.61 (d, 2H, J = 3.2 Hz), 7.14 (t, 2H, J = 8.8 Hz), 2.51 (s, 6H). 13C NMR (100 MHz, DMSO-d6, ppm): 159.44 (C=O), 154.55 (Ar-C), 148.91 (Ar-C), 143.22 (-C=N-), 140.48 (Ar-C), 130.06 (Ar-C), 129.44 (Ar-C), 128.20 (Ar-C), 125.75 (Ar- C), 15.32 (-CH3).	2	1	Because, their polyfunctional nature offer multifarious synthetic ways to derivatize such organic molecules towards suitable and effective drug-receptor interaction.			molecules27051656_makarova	0
+We appreciate the recommendation and agree with the reviewer that the computational and experimental results of antimicrobial activity differ to some extent in this part of the study and required further investigation to understand why such activity differs to the ligands by other means in the future. In this study, we successfully synthesized the novel ligands and studied its spectroscopy with other probable studies. Fortunately, we have found outstanding results during the chemo-sensor study and hopefully, we could be able to show the excellent usage of the ligands as a chemo-sensor in near future. Also, we are planning to do a fluorescence imaging study of protein-ligands.	2	1	Ligand and Protein preparation: The structures of L1 and L2 have been fully optimized by using Gaussian 09 software at B3LYP/6-311G+ (d, p) level.			molecules27051656_makarova	0
+We appreciate the comments of the reviewer and the following are the answers in response to the question for the kind consideration of the reviewer- Tyrosinase is a key enzyme in melanogenesis, which is essential for pigmentation. Dysfunction of tyrosinase may cause skin cancer. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-103]. That’s why, to investigate and compare the antimicrobial activity of the synthesized compounds with in vitro data, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium was performed. Also, molecular docking with other proteins of B. Megaterium was conducted, but 46ju provided better binding affinity with the synthesized compounds. For this reason, we aim to study the structure of tyrosinase from the bacteria. Preliminary antimicrobial studies were carried out to validate the docking protocol. At present, we are doing the fluorescence activities of these ligands and L2 showed excellent fluorescent emission at λmax 520 nm. So, we hope that we will carry out ALA scan (Alanine scanning) based MD experiments and 100ns Molecular dynamics (MD) simulation guided protein-ligand stability in the future for our next study since within this shortest time, we are unable to do these experiments. The active binding site of tyrosinase was predicted by CASTp. We have included this in the manuscript as follows: Binding affinity of L1 and L2 The highest anti-bacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in the Fig. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala44A, and ALA44B of tyrosinase enzyme (PDB ID: 4j6u), which also supports the literature [101-103]. Thus, computational results are in good agreement with in vitro antibacterial behaviour of our compounds for novel antibacterial drug design.	2	1	Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96].			molecules27051656_makarova	0
+We are delighted and thankful for the recommendation of the reviewer.	4	1	[31] Ermiş, E. Synthesis, spectroscopic characterization and DFT calculations of novel Schiff base containing thiophene ring.			molecules27051656_makarova	0
+We are delighted and thankful for the recommendation.	2	1	It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].			molecules27051656_makarova	0
+We appreciate the recommendation and rewrite the abstract as follows, Polytopic organic ligands with hydrazone moiety are in the forefront of new drug research among many others due to their unique and versatile functionality and ease of strategic ligand design. Quantum chemical calculations of these polyfunctional ligands can be carried out in silico to determine the thermodynamic parameters. In this report two new tritopic dihydrazide ligands, N'2, N'6-bis[(1E)-1-(thiophen-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L1) and N'2, N'6-bis[(1E)-1-(1H-pyrrol-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L2) were successfully prepared by the condensation reaction of pyridine-2, 6-dicarboxylic hydrazide with 2-acetylthiophene and 2-acetylpyrrole. The FT-IR, 1H and 13C NMR as well as mass spectra of both L1 and L2 were recorded and analyzed. Quantum chemical calculations were performed at DFT/B3LYP/cc-pvdz/6-311+ G (d, p) level of theory to study the molecular geometry, vibrational frequencies, and thermodynamic properties including changes of ∆H, ∆S, and ∆G for both the ligands. The optimized vibrational frequency and (1H and 13C) NMR obtained by B3LYP/cc-pvdz/6-311 + G (d, p) showed good agreement with experimental FT-IR and NMR data. Frontier molecular orbital (FMO) calculations were also conducted to find the HOMO, LUMO, and HOMO–LUMO gaps of the two synthesized compounds. To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain. In addition, Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium. The outcomes revealed that both L1 and L2 can suppress microbial growth of bacteria and fungi with variable potency. The antibacterial activity results demonstrated the compound L2 to be potentially effective against Bacillus Megaterium with inhibition zones of 12 mm while molecular docking study showed the binding energies for L1 and L2 to be −7.7 and −8.8 kcal mol−1 respectively with tyrosinase from Bacillus megaterium.‪ -	2	1	Response: We appreciate the recommendation and changed the conclusion as follows, 9 Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized to present a wide range of biological activities in recent years.			molecules27051656_makarova	0
+We appreciate the recommendation and corrected the font of the references.	2	1	After incubation, the test organism was inoculated using sterile cotton bar on media.			molecules27051656_makarova	0
+We appreciate the recommendation and corrected the section as below: Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development [1-6]. Because, their polyfunctional nature offer multifarious synthetic ways to derivatize such organic molecules towards suitable and effective drug-receptor interaction. The derivatives of hydrazide-hydrazone moiety specially with heterocyclic system possess a range of biological activities namely, anti-microbial, anti-mycobacterial, antitubercular, anticonvulsant, anticholinesterase [1], antiplatelet, and more importantly antitumor [5,7]. Transition metal complexes derived from such type of ligands have been widely studied since they also demonstrate significant biological and pharmacological properties [8-11].	2	1	[25] Jagtap, V. A.; Agasimundin, Y. S. Synthesis and preliminary evaluation of some 2-amino-N'-[substituted]-4,5,6,7-tetrahydro-1-benzothiophene-3-carbohydrazide as antimicrobial agents.			molecules27051656_makarova	0
+We appreciate the recommendation and corrected the section as below: This Section is corrected as “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.’’	2	1	Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40].			molecules27051656_makarova	0
+We appreciate the recommendation and added the required references as below. We have these specimens of strains available in our lab to study. Therefore, we used. We also updated the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B and corrected the reference the number of Salmonella Typhi (K-323130) bacteria. We are showing our sincere apology for such mistakes. The changes are added as follows,	2	1	Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl).			molecules27051656_makarova	0
+4.9. We appreciate the recommendation, and corrected this information as shown below: 3.2.	2	1	[63] Silverstein, R. M.; Webster, F. X. Spectroscopic Identification of Organic Compound, sixth ed., John Willey & Sons, New York, 1998.			molecules27051656_makarova	0
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation. Protein-ligand Docking 3.2.1. Ligand and Protein preparation: The structures of L1 and L2 have been fully optimized by using Gaussian 09 software at B3LYP/6-311G+ (d, p) level. The 3D crystal structure of tyrosinase from Bacillus Megaterium (PDB ID: 4j6u; resolution: 2.5Å, Chain A, B) was obtained in pdb format from online RCSB protein data bank (PDB) database. The structure was verified, and an energy minimization was performed with the Swiss-Pdb Viewer software packages (version 4.1.0) [44], since the crystal structure contains a variety of issues related to improper bond order, side chains geometry, and missing hydrogen atoms. Prior to docking, all the heteroatoms and water molecules were removed from the crystal structure using PyMol (version 1.3) software packages [45]. The active binding pocket of tyrosinase was predicted by CASTp—having the highest pocket area and volume are 95.432 Å2 and 137.877 Å3, respectively [46]. Both the structures of the proteins and ligands were saved in .pdbqt format by AutoDock Vina (version 1.1.2, May 11, 2011) for docking analysis [47]. 3.2.2. Molecular docking Analysis: The docking calculations were performed using default parameters and 8 docked conformations were generated for both compounds. The energy calculations were done by genetic algorithms. Nonpolar hydrogen atoms, Gasteiger partial charges, rotatable bonds, and grid box with dimensions 66.57 × 58.25 × 84.98 Å3 created on the tyrosinase with the aid of Auto Dock Tools 1.1.2 and spacing of 0.3750 Å. The docked conformation of the respective protein conformer with lowest binding free energy and root mean-square deviation value (RMSD) 0.0 Å was analyzed using PyMOL Molecular Graphics System (version 1.7.4) and Accelrys Discovery Studio 4.1 [49]. We appreciate the recommendation, and splitted the data into 2 more subheads as, 3.2.1. Ligand and Protein preparation and 3.2.2. Molecular docking analysis. Also, more information was added about the software used as shown below: 3.2.	2	1	The conclusion must remind the reader why the article was written in the first place and why it is important in the field.			molecules27051656_makarova	0
+2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study. We appreciate the recommendation, and this section is updated by changing the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B. Also, the sentences of second part (4.8) are deleted and added to 2.3.1 as suggested by the reviewer. Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method. The formation of diameter of inhibition zones in mm by the synthesized analogues are shown in Table 5. Compound L2 showed moderate activity against Bacillus Megaterium bacteria while L1 showed promising antifungal activity against Aspergillus niger fungal strains compared to standard Amphotericin-B. Diameter of inhibition zones (mm) of the synthesized compounds, Ceftriaxone and Amphotericin-B against tested bacterial and fungal strains.	2	1	As they did not report any enzyme specific inhibition related experiments.			molecules27051656_makarova	0
+Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100]. We appreciate the recommendation and updated the molecular docking study by adding more details and literature references as follows, 4.9.	2	1	The active binding site of tyrosinase was predicted by CASTp.			molecules27051656_makarova	0
+We appreciate the recommendation and changed the conclusion as follows, Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized to present a wide range of biological activities in recent years. In this present study we have synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and mass spectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also done. The computed FT-IR analysis as well as the 1H and 13C NMR using B3LYP/CC-PVDZ/6-311+G(d, p) method agreed satisfactorily with the experimental results. We further evaluated the thermodynamic parameters ∆H, ∆S, and ∆G of the ligands. The geometry optimization revealed the planarity of L1 and L2 molecules. Further, it was seen from the HOMO-LUMO energy values that the chemical potentials were negative and the frontier orbital gap of the molecule under investigation was small, and hence, both compounds are reactive and polarizable. To further showcase the biological activity of the ligands against organic pathogens, the antimicrobial assay was performed and revealed significant inhibition of L2 against Bacillus Megaterium gram positive bacteria, and L1 against Escherichia coli, Aspergillus niger although in lesser extent. The moderate activity of L2 molecule against Bacillus Megaterium is substantiated by molecular docking study against tyrosinase from Bacillus megaterium and was found significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which might suggest the antimicrobial activity of the molecule. Overall, L1 and L2 compounds have spurred significant interest for us from the synthetic, computational and biological point of view. We anticipate continued research regarding these classes of exciting organic ligands.	2	1	Response: We appreciate the recommendation and updated the molecular docking study by adding more details and literature references as follows, 4.9.			molecules27051656_makarova	0
+4.9.1. 6. In L1-4j6u, one conventional hydrogen bond (3.04 Å) of O-H----O-C observed between O-H of Tyr267A and O-C group of compounds L1. Pi-cation, pi-sulfur and amide-pi bonds were also noticed with LYS47A, PHE48A, ILE39A, ALA40A, GLY43B and ALA44B, respectively. Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl). L2-4j6u complex was stabilized by four NH….O hydrogen bonds and they were LYS47A (2.25 Å), GLY143B (3.04 Å), Tyr267A (3.07 Å) and PRO219B (2.91 Å) (Fig. 6). Like L1, L2 formed pi-cation and amide-pi bonds with LYS47A, ILE39A, GLY43B, where the distances were 3.73, 4.34, 3.54 Å. L2 also formed seven pi-alkyl bonds with ALA44A (5.19 Å), LYS47A (4.35 Å), ALA44B (3.95 Å), LYS47B (4.98 Å), PRO52A (5.04 Å), ALA40B (4.67 Å) and ILE139B (4.24 Å), respectively. We appreciate the recommendation and rewrote the Molecular Docking study as follows as suggested by the reviewer, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding of ligand-receptor interactions in order to facilitate the design of potential drugs [92-95]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analyses of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [96-99]. The docking results were also compared with well-testified inhibitor arbutin [97]. Binding affinity of L1 and L2 The highest antibacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, whereas for arbutin-4j6u the value was -9.1 kcal mol-1 which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in Fig. It was observed that arbutin formed six conventional hydrogen bonds with 4j6u (Supplementary Fig. S11) by the following residues: ALA40A (one O-H----O-C hydrogen bond), Glu141A (four O-H----O-C hydrogen bond), and LYS47B (one O-H----O-C hydro-gen bond). Also several hydrophobic interactions were found with ILE139A, ILE39A, and ALA40A. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala40A, Ala44A, ALA44B and Lys47B of tyrosinase enzyme (PDB ID: 4j6u), which are in close vicinity to the control arbutin and supports the literature [100-102].	4	1	- Replace “In addition to that, the compounds were tested in vitro bioassays against some bacteria and the fungus strain showing specially promising positive - negative and Gram - Gram odology was used to study molecular behavior of L1 and L2 results for L2.			molecules27051656_makarova	0
+We appreciate the recommendation and corrected the style of the references as suggested by the reviewer.	4	1	[77] Gökce, H.; Öztürk, N.; Kazıcı, M.; Yörür Göreci, Ç.; Güneş, S. Structural, spectroscopic, electronic, nonlinear optical and thermodynamic properties of a synthesized Schiff base compound: A combined experimental and theoretical approach.			molecules27051656_makarova	0
+We appreciate the recommendation and comments. We corrected the above-mentioned correction as suggested by the reviewer and presented as follows, 1H NMR (400 MHz, DMSO-d6, ppm): δ 11.43 (s, 2H), 8.34 (m, 2H), 8.28 (m, 1H), 7.65 (d, 2H, J = 4.8 Hz), 7.61 (d, 2H, J = 3.2 Hz), 7.14 (t, 2H, J = 8.8 Hz), 2.51 (s, 6H). 13C NMR (100 MHz, DMSO-d6, ppm): 159.44 (C=O), 154.55 (Ar-C), 148.91 (Ar-C), 143.22 (-C=N-), 140.48 (Ar-C), 130.06 (Ar-C), 129.44 (Ar-C), 128.20 (Ar-C), 125.75 (Ar- C), 15.32 (-CH3).	2	1	After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked.			molecules27051656_perova	0
+We appreciate the recommendation and agree with the reviewer that the computational and experimental results of antimicrobial activity differ to some extent in this part of the study and required further investigation to understand why such activity differs to the ligands by other means in the future. In this study, we successfully synthesized the novel ligands and studied its spectroscopy with other probable studies. Fortunately, we have found outstanding results during the chemo-sensor study and hopefully, we could be able to show the excellent usage of the ligands as a chemo-sensor in near future. Also, we are planning to do a fluorescence imaging study of protein-ligands.	2	1	10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively.			molecules27051656_perova	0
+We appreciate the comments of the reviewer and the following are the answers in response to the question for the kind consideration of the reviewer- Tyrosinase is a key enzyme in melanogenesis, which is essential for pigmentation. Dysfunction of tyrosinase may cause skin cancer. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-103]. That’s why, to investigate and compare the antimicrobial activity of the synthesized compounds with in vitro data, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium was performed. Also, molecular docking with other proteins of B. Megaterium was conducted, but 46ju provided better binding affinity with the synthesized compounds. For this reason, we aim to study the structure of tyrosinase from the bacteria. Preliminary antimicrobial studies were carried out to validate the docking protocol. At present, we are doing the fluorescence activities of these ligands and L2 showed excellent fluorescent emission at λmax 520 nm. So, we hope that we will carry out ALA scan (Alanine scanning) based MD experiments and 100ns Molecular dynamics (MD) simulation guided protein-ligand stability in the future for our next study since within this shortest time, we are unable to do these experiments. The active binding site of tyrosinase was predicted by CASTp. We have included this in the manuscript as follows: To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100]. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in the Fig. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala44A, and ALA44B of tyrosinase enzyme (PDB ID: 4j6u), which also supports the literature [101-103]. Thus, computational results are in good agreement with in vitro antibacterial behaviour of our compounds for novel antibacterial drug design.	2	1	Response: We appreciate the recommendation and comments.			molecules27051656_perova	0
+We are delighted and thankful for the recommendation.	2	1	A.; Shchekotikhin, A. E. Amides of pyrrole- and thiophene-fused anthraquinone derivatives: A role of the heterocyclic core in antitumor properties.			molecules27051656_perova	0
+We appreciate the recommendation and rewrite the abstract as follows, Polytopic organic ligands with hydrazone moiety are in the forefront of new drug research among many others due to their unique and versatile functionality and ease of strategic ligand design. Quantum chemical calculations of these polyfunctional ligands can be carried out in silico to determine the thermodynamic parameters. In this report two new tritopic dihydrazide ligands, N'2, N'6-bis[(1E)-1-(thiophen-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L1) and N'2, N'6-bis[(1E)-1-(1H-pyrrol-2-yl) ethylidene] pyridine-2, 6-dicarbohydrazide (L2) were successfully prepared by the condensation reaction of pyridine-2, 6-dicarboxylic hydrazide with 2-acetylthiophene and 2-acetylpyrrole. The FT-IR, 1H and 13C NMR as well as mass spectra of both L1 and L2 were recorded and analyzed. Quantum chemical calculations were performed at DFT/B3LYP/cc-pvdz/6-311+ G (d, p) level of theory to study the molecular geometry, vibrational frequencies, and thermodynamic properties including changes of ∆H, ∆S, and ∆G for both the ligands. The optimized vibrational frequency and (1H and 13C) NMR obtained by B3LYP/cc-pvdz/6-311 + G (d, p) showed good agreement with experimental FT-IR and NMR data. Frontier molecular orbital (FMO) calculations were also conducted to find the HOMO, LUMO, and HOMO–LUMO gaps of the two synthesized compounds. To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain. In addition, Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium. The outcomes revealed that both L1 and L2 can suppress microbial growth of bacteria and fungi with variable potency. The antibacterial activity results demonstrated the compound L2 to be potentially effective against Bacillus Megaterium with inhibition zones of 12 mm while molecular docking study showed the binding energies for L1 and L2 to be −7.7 and −8.8 kcal mol−1 respectively with tyrosinase from Bacillus megaterium.‪ -	2	1	The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis.			molecules27051656_perova	0
+We appreciate the recommendation and corrected the font of the references.	2	1	1 Manuscript ID #: Molecules 1583723 Journal: Molecules A detailed point-by-point response to the reviewers' comments We would like to thank the reviewers for the comments and endorsements of our work.			molecules27051656_perova	0
+We appreciate the recommendation and corrected the section as below: Polytopic ligands containing hydrazide-hydrazone moiety (—CO—NHN=CH—) are important for new drug development [1-6]. Because, their polyfunctional nature offer multifarious synthetic ways to derivatize such organic molecules towards suitable and effective drug-receptor interaction. The derivatives of hydrazide-hydrazone moiety specially with heterocyclic system possess a range of biological activities namely, anti-microbial, anti-mycobacterial, antitubercular, anticonvulsant, anticholinesterase [1], antiplatelet, and more importantly antitumor [5,7]. Transition metal complexes derived from such type of ligands have been widely studied since they also demonstrate significant biological and pharmacological properties [8-11].	2	1	[8] Zhao, L.; Xu, Z.; Thompson, L. K.; Heath, S. L.; Miller, D. O.; Ohba,.			molecules27051656_perova	0
+We appreciate the recommendation and corrected the section as below: This Section is corrected as “‪In addition to that, the compounds were tested in vitro against Gram-negative and positive ‪bacteria and two fungi. Molecular ‪docking was used to study the molecular behavior of L1 and L2 against tyrosinase from Bacillus megaterium.’’ Section 2.3.1.	2	1	The energy calculations were done by genetic algorithms.			molecules27051656_perova	0
+We appreciate the recommendation and added the required references as below. We have these specimens of strains available in our lab to study. Therefore, we used. We also updated the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B and corrected the reference the number of Salmonella Typhi (K-323130) bacteria. We are showing our sincere apology for such mistakes. The changes are added as follows, 2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study.	2	1	We also updated the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B and corrected the reference the number of Salmonella Typhi (K-323130) bacteria.			molecules27051656_perova	0
+4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. We appreciate the recommendation, and corrected this information as shown below: 3.2.	2	1	[40] Balouiri, M.; Sadiki, M.; Ibnsouda, S. K. Methods for in vitro evaluating antimicrobial activity: A review.			molecules27051656_perova	0
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation. Protein-ligand Docking 3.2.1. Ligand and Protein preparation: The structures of L1 and L2 have been fully optimized by using Gaussian 09 software at B3LYP/6-311G+ (d, p) level. The 3D crystal structure of tyrosinase from Bacillus Megaterium (PDB ID: 4j6u; resolution: 2.5Å, Chain A, B) was obtained in pdb format from online RCSB protein data bank (PDB) database. The structure was verified, and an energy minimization was performed with the Swiss-Pdb Viewer software packages (version 4.1.0) [44], since the crystal structure contains a variety of issues related to improper bond order, side chains geometry, and missing hydrogen atoms. Prior to docking, all the heteroatoms and water molecules were removed from the crystal structure using PyMol (version 1.3) software packages [45]. The active binding pocket of tyrosinase was predicted by CASTp—having the highest pocket area and volume are 95.432 Å2 and 137.877 Å3, respectively [46]. Both the structures of the proteins and ligands were saved in .pdbqt format by AutoDock Vina (version 1.1.2, May 11, 2011) for docking analysis [47]. 3.2.2. Molecular docking Analysis: The docking calculations were performed using default parameters and 8 docked conformations were generated for both compounds. The energy calculations were done by genetic algorithms. Nonpolar hydrogen atoms, Gasteiger partial charges, rotatable bonds, and grid box with dimensions 66.57 × 58.25 × 84.98 Å3 created on the tyrosinase with the aid of Auto Dock Tools 1.1.2 and spacing of 0.3750 Å. The docked conformation of the respective protein conformer with lowest binding free energy and root mean-square deviation value (RMSD) 0.0 Å was analyzed using PyMOL Molecular Graphics System (version 1.7.4) and Accelrys Discovery Studio 4.1 [49]. We appreciate the recommendation, and splitted the data into 2 more subheads as, 3.2.1. Ligand and Protein preparation and 3.2.2. Molecular docking analysis. Also, more information was added about the software used as shown below: 3.2.	2	1	[72] Azad, I.; Akhter, Y.; Khan, T.; Azad, M. I.; Chandra, S.; Singh, P.; Kumar, D.; Nasibullah, M. Synthesis, quantum chemical study, AIM simulation, in silico ADMET profile analysis, molecular docking and antioxidant activity assessment of aminofuran derivatives.			molecules27051656_perova	0
+We appreciate the recommendation, and this section is updated by changing the standard ciprofloxacin and miconazole with Ceftriaxone and Amphotericin-B. Also, the sentences of second part (4.8) are deleted and added to 2.3.1 as suggested by the reviewer. 2.1.3. Antimicrobial activity assay In vitro antimicrobial activity of synthesized ligands was evaluated by agar disc diffusion method [40]. Mueller Hinton Agar (MHA) media (HIMEDIA, India) was used as a control medium for testing against bacteria and Potato Dextrose Agar (PDA) media (HIMEDIA, India) was used for fungal strain. After preparation, the MHA and PDA medias were incubated for 24 h and contaminations were checked. After incubation, the test organism was inoculated using sterile cotton bar on media. The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay. Dimethyl sulfoxide (DMSO) was used as control. Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds. 10 µL of ceftriaxone and amphotericin-B solutions containing 50 µg each in DMSO were loaded per disc for antibacterial and antifungal assays as positive control, respectively. The diameter of the inhibition zones in mm circling the disc were measured. Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study. 4.8. Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method. The formation of diameter of inhibition zones in mm by the synthesized analogues are shown in Table 5. Compound L2 showed moderate activity against Bacillus Megaterium bacteria while L1 showed promising antifungal activity against Aspergillus niger fungal strains compared to standard Amphotericin-B. Table 5. Diameter of inhibition zones (mm) of the synthesized compounds, Ceftriaxone and Amphotericin-B against tested bacterial and fungal strains.	2	1	[33] Aljahdali, M. S.; Abdou El-Sherif, A.; Hilal, R.H.; Abdel-Karim, A.T. Mixed bivalent transition metal complexes of 1,10-phenanthroline and 2-aminomethylthiophenyl-4-bromo salicyl aldehyde Schiff base: Spectroscopic, molecular modeling and biological activities.			molecules27051656_perova	0
+We appreciate the recommendation and updated the molecular docking study by adding more details and literature references as follows, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].	2	1	The binding site residues predicted by CASTp for tyrosinase were used for grid generation.			molecules27051656_perova	0
+In this present study we have synthesized two tritopic dihydrazide based ligands bearing Pyrrole and Thiophene as end groupings and characterized successfully by FT-IR, 1H and 13C NMR and mass spectrometry. Based on the DFT the calculations, a complete structural detail, vibrational, electrostatic potential, Mulliken population, HOMO-LUMO and thermodynamic analysis were also done. The computed FT-IR analysis as well as the 1H and 13C NMR using B3LYP/CC-PVDZ/6-311+G(d, p) method agreed satisfactorily with the experimental results. We further evaluated the thermodynamic parameters ∆H, ∆S, and ∆G of the ligands. The geometry optimization revealed the planarity of L1 and L2 molecules. Further, it was seen from the HOMO-LUMO energy values that the chemical potentials were negative and the frontier orbital gap of the molecule under investigation was small, and hence, both compounds are reactive and polarizable. To further showcase the biological activity of the ligands against organic pathogens, the antimicrobial assay was performed and revealed significant inhibition of L2 against Bacillus Megaterium gram positive bacteria, and L1 against Escherichia coli, Aspergillus niger although in lesser extent. The moderate activity of L2 molecule against Bacillus Megaterium is substantiated by molecular docking study against tyrosinase from Bacillus megaterium and was found significant with a binding energy of -8.8 kcal/mol and three hydrogen bond interactions, which might suggest the antimicrobial activity of the molecule. Overall, L1 and L2 compounds have spurred significant interest for us from the synthetic, computational and biological point of view. We anticipate continued research regarding these classes of exciting organic ligands. We appreciate the recommendation and changed the conclusion as follows, 9 Pyrrole and thiophene as organic molecules and their metal cluster derivatives have been recognized to present a wide range of biological activities in recent years.	2	1	"explored ""Synthesis of Novel Tritopic Hydrazone Ligands: Spectroscopy, Biological activity, DFT, and Molecular docking Studies""."			molecules27051656_perova	0
+4.9.1. Binding affinity of L1 and L2 The highest anti-bacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. 6. In L1-4j6u, one conventional hydrogen bond (3.04 Å) of O-H----O-C observed between O-H of Tyr267A and O-C group of compounds L1. Pi-cation, pi-sulfur and amide-pi bonds were also noticed with LYS47A, PHE48A, ILE39A, ALA40A, GLY43B and ALA44B, respectively. Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl). L2-4j6u complex was stabilized by four NH….O hydrogen bonds and they were LYS47A (2.25 Å), GLY143B (3.04 Å), Tyr267A (3.07 Å) and PRO219B (2.91 Å) (Fig. 6). Like L1, L2 formed pi-cation and amide-pi bonds with LYS47A, ILE39A, GLY43B, where the distances were 3.73, 4.34, 3.54 Å. L2 also formed seven pi-alkyl bonds with ALA44A (5.19 Å), LYS47A (4.35 Å), ALA44B (3.95 Å), LYS47B (4.98 Å), PRO52A (5.04 Å), ALA40B (4.67 Å) and ILE139B (4.24 Å), respectively. We appreciate the recommendation and rewrote the Molecular Docking study as follows as suggested by the reviewer, 4.9. Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding of ligand-receptor interactions in order to facilitate the design of potential drugs [92-95]. To investigate and compare the antimicrobial activity of the synthesized compounds, docking analyses of L1 and L2 against tyrosinase from Bacillus Megaterium were performed. It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [96-99]. The docking results were also compared with well-testified inhibitor arbutin [97]. Binding affinity of L1 and L2 The highest antibacterial activity (zone of inhibition 12 mm) of compound L2 was detected with tyrosinase from Bacillus Megaterium (PDB ID: 4j6u) bacteria compared to L1. The binding energies for L1 and L2 with Bacillus Megaterium were −7.7 and −8.8 kcal mol−1, respectively, whereas for arbutin-4j6u the value was -9.1 kcal mol-1 which were calculated by AutoDock Vina. The interactions of the 4j6u with compounds L1 and L2 are shown in Fig. It was observed that arbutin formed six conventional hydrogen bonds with 4j6u (Supplementary Fig. S11) by the following residues: ALA40A (one O-H----O-C hydrogen bond), Glu141A (four O-H----O-C hydrogen bond), and LYS47B (one O-H----O-C hydro-gen bond). Also several hydrophobic interactions were found with ILE139A, ILE39A, and ALA40A. Results of docking studies revealed that L1 and L2 formed bonds to the active site of tyrosinase and showed strong interactions with Tyr267A, Ala40A, Ala44A, ALA44B and Lys47B of tyrosinase enzyme (PDB ID: 4j6u), which are in close vicinity to the control arbutin and supports the literature [100-102].	4	1	The diameter of the inhibition zones in mm circling the disc were measured.			molecules27051656_perova	0
+We appreciate the recommendation and corrected the style of the references as suggested by the reviewer.	4	1	[6] Shebl, M. Coordination behavior of new bis(tridentate ONO, ONS and ONN) donor hydrazones towards some transition metal ions: Synthesis, spectral, thermal, antimicrobial and antitumor studies.			molecules27051656_perova	0
+We have changed the position of Figure 1 to line 182, and adjusted the arrangement of the figures in the article.	2	1	The point-by-point responses to the comments are enclosed.			molecules27123843_makarova	0
+We have removed the heading “2. Results” in the revised manuscript.	2	1	Reviewer's comments and authors' replies: [Reviewer comments in black; Author response in blue.]			molecules27123843_makarova	0
+Thank you for your valuable opinion. NAA is an abbreviation for amino acid, and we have added the description of NAA in the section of “2.3 Construction of sensor array”.	2	1	We truly appreciate your positive comments on our work, as well as for raising interesting points, which lead to the improvement of the manuscript.			molecules27123843_makarova	0
+We have made the appropriate changes in the text to make the sentences more concise and highlighted them in red (line 248).	2	1	Author Response Author Response File: Author Response.docx			molecules27123843_makarova	0
+Thanks for the valuable comment. We have removed the word “that” in the revised manuscript.	2	1	Reviewer's comments and authors' replies: [Reviewer comments in black; Author response in blue.]			molecules27123843_makarova	0
+We have changed the position of Figure 1 to line 182, and adjusted the arrangement of the figures in the article.	2	1	The point-by-point responses to the comments are enclosed.			molecules27123843_perova	0
+We have removed the heading “2. Results” in the revised manuscript.	2	1	Although the method of preparation of these sensors is not new, the implementation to chemosensor array is well presented.			molecules27123843_perova	0
+Thank you for your valuable opinion. NAA is an abbreviation for amino acid, and we have added the description of NAA in the section of “2.3 Construction of sensor array”.	2	1	It is also impressive that all 11 metal ions were successfully identified by LDA with 100% classification rate.			molecules27123843_perova	0
+We have made the appropriate changes in the text to make the sentences more concise and highlighted them in red (line 248).	2	1	Author Response Author Response File: Author Response.docx			molecules27123843_perova	0
+Thanks for the valuable comment. We have removed the word “that” in the revised manuscript.	2	1	Results”, but the content of that paragraph belong to the introduction part.			molecules27123843_perova	0
+Our studies have shown that the effect of metal vapors on the color of the discharge plasma is much greater in the case of a pulsed breakdown in a nonuniform electric field. This made it possible to compare with the color of various discharges occurring in the upper atmosphere (Figure 1 in the text of the article). Even with well-studied discharges, it is very difficult to obtain such results. Thus, during an arc discharge, although there is a strong evaporation of the electrodes and the ejection of particles of various sizes, the voltage across the gap is low and the discharge in metal vapors outside the high-temperature arc channel is practically absent. Under these conditions, broadband Planck radiation from the arc channel plasma is observed. In a glow discharge, the sputtering of the electrodes has a low rate, and the emission spectra of the plasma of such discharges contain atomic and molecular transitions of the gases used. In the investigated mode, metal vapors are produced and excited by a pulsed discharge.	2	1	Please describe better your experimental system, such as ignition energy.			nano12040652_makarova	0
+Plasma concentration discussions have been removed from the text because no such measurements have been made. The nature of the glow of the particles in Figure 3 corresponds to the glow of a micrometeorite that burns down in the Earth's atmosphere [13] and [http://galaxy.astron.kharkov.ua/statti/meteor.htm]. The brightness of the micrometeorite (the particle) glow increases towards the track’s end. This cannot be explained by an increase in the particle velocity, since the particle stops. This occurs after the voltage pulse action. We believe that an increase in the radiation intensity of the particle is due to its heating during deceleration on gas particles. The text of the article has been revised. Such inaccuracies are found throughout the text.	2	1	"For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."""			nano12040652_makarova	0
+The text of the article was finalized and the English was improved.	2	1	Under these conditions, broadband Planck radiation from the arc channel plasma is observed.			nano12040652_makarova	0
+The introduction of the manuscript and its text have been revised. In addition, new references have been added to the article. All changes in the text are highlighted in either yellow or red font. First of all, data describing the characteristics of the pulsed nanosecond discharge used were added and the influence of the electrode material was described in more detail. On the other hand, we should note that with a voltage pulse duration from ones to tens of nanoseconds, the experimental conditions differ significantly from the conditions of an electrostatic discharge and the creation of a foggy environment.	2	1	Edits in the text are highlighted in either yellow or red font.			nano12040652_makarova	0
+The experimental system was described in more detail, and various modes of discharge ignition were analyzed.	2	1	In addition, there are comments on the research methodology and interpretation of research on the article itself.			nano12040652_makarova	0
+In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence. Since under these conditions diffuse discharges, at which the voltage across the gap remains high are formed, metal vapors are excited and ionized together with gas molecules in the discharge gap. This leads to the emission of radiation at various spectral transitions of metal atoms. Only a part of these transitions has a high radiation intensity in the region of interest to researchers. Also, to obtain a high intensity of radiation, transitions of atoms in metal vapors can be used, which are populated as a result of the efficient transfer of energy from excited gas molecules and atoms. Metals, the color of the emission of vapors of which, when excited in the plasma of nanosecond discharges, corresponds to the color of high-altitude atmospheric discharges, were chosen as the material of the electrodes.	2	1	Please describe better your experimental system, such as ignition energy.			nano12040652_makarova	0
+The color of the glow of the discharge plasma at electrodes made of various metals is associated with excitation certain energy levels of particles in the vapors of these metals. So, when using electrodes made of aluminum, steel and copper, we observed the glow of red, blue and green colors, respectively. The different colors of the discharge when changing the material of the electrodes are determined not by the spark or arc stages, but by bright spots on the electrodes, which are formed due to the explosive emission of electrons [Mesyats, G.A. Ecton mechanism of the vacuum arc cathode spot. IEEE transactions on plasma science, 1995, 23(6), pp. 879-883. (DOI: 10.1109/27.476469)]. These areas in the photographs have a bright white color (see, for example, the photographs in Figures 3, 6, 7). In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material. High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap. However, emission of individual particles is determined by their temperature, it corresponds to the Planck radiation and is broadband. We note once again that in this work, to obtain metal vapors, as well as metal nano- and microparticles, a pulsed nanosecond discharge in a non-uniform electric field was used.	2	1	One can agree with the influence of the particle size on the track size, but how can an increase in the particle charge affect the track glow?			nano12040652_makarova	0
+Several references was added.	2	1	Some data should be provided to show the elemental composition of the particles.			nano12040652_makarova	0
+The main experiments were carried out with discharges in air, and the air pressure was chosen close to the pressures of high-altitude discharges. Other gases, such as argon, were chosen to better demonstrate the discoloration of the discharge.	2	1	The introduction would benefit from adding more content and detailed review overview to this research, such as particle lifting in electrostatic discharge, Turbulence effect, Mist-containing environment, Electrode materials.			nano12040652_makarova	0
+The manuscript has been revised. This judgment was removed from the text of the manuscript.	2	1	We hope that the results presented in the article will also be of interest to researchers who deal with micro and nanoparticles in traditional areas of nanomaterial physics [1-8].			nano12040652_makarova	0
+Thanks for the advice. We plan to do this in our next work.	2	1	Reply to Reviewer 2 The article: «Influence of nanoparticles and metal vapors on the color of laboratory and atmospheric discharges» Authors: Victor Tarasenko, Nikita Vinogradov, Dmitry Beloplotov, Alexander Burachenko, Mikhail Lomaev, and Dmitry Sorokin Submission Date: 14 December 2021 Date of this review: 19 Jan 2022 07:18:39 Comments and Suggestions for Authors 1.			nano12040652_makarova	0
+Our studies have shown that the effect of metal vapors on the color of the discharge plasma is much greater in the case of a pulsed breakdown in a nonuniform electric field. This made it possible to compare with the color of various discharges occurring in the upper atmosphere (Figure 1 in the text of the article). Even with well-studied discharges, it is very difficult to obtain such results. Thus, during an arc discharge, although there is a strong evaporation of the electrodes and the ejection of particles of various sizes, the voltage across the gap is low and the discharge in metal vapors outside the high-temperature arc channel is practically absent. Under these conditions, broadband Planck radiation from the arc channel plasma is observed. In a glow discharge, the sputtering of the electrodes has a low rate, and the emission spectra of the plasma of such discharges contain atomic and molecular transitions of the gases used. In the investigated mode, metal vapors are produced and excited by a pulsed discharge.	2	1	Edits in the text are highlighted in either yellow or red font.			nano12040652_perova	0
+Plasma concentration discussions have been removed from the text because no such measurements have been made. The nature of the glow of the particles in Figure 3 corresponds to the glow of a micrometeorite that burns down in the Earth's atmosphere [13] and [http://galaxy.astron.kharkov.ua/statti/meteor.htm]. The brightness of the micrometeorite (the particle) glow increases towards the track’s end. This cannot be explained by an increase in the particle velocity, since the particle stops. This occurs after the voltage pulse action. We believe that an increase in the radiation intensity of the particle is due to its heating during deceleration on gas particles. The text of the article has been revised.	2	1	Reply: This judgment was removed from the text of the manuscript.			nano12040652_perova	0
+The text of the article was finalized and the English was improved.	2	1	The topic is timely and the results provided are interesting.			nano12040652_perova	0
+The introduction of the manuscript and its text have been revised. In addition, new references have been added to the article. All changes in the text are highlighted in either yellow or red font. First of all, data describing the characteristics of the pulsed nanosecond discharge used were added and the influence of the electrode material was described in more detail. On the other hand, we should note that with a voltage pulse duration from ones to tens of nanoseconds, the experimental conditions differ significantly from the conditions of an electrostatic discharge and the creation of a foggy environment.	2	1	This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc.			nano12040652_perova	0
+The experimental system was described in more detail, and various modes of discharge ignition were analyzed.	2	1	Reply: Plasma concentration discussions have been removed from the text because no such measurements have been made.			nano12040652_perova	0
+In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence. Since under these conditions diffuse discharges, at which the voltage across the gap remains high are formed, metal vapors are excited and ionized together with gas molecules in the discharge gap. This leads to the emission of radiation at various spectral transitions of metal atoms. Only a part of these transitions has a high radiation intensity in the region of interest to researchers. Also, to obtain a high intensity of radiation, transitions of atoms in metal vapors can be used, which are populated as a result of the efficient transfer of energy from excited gas molecules and atoms. Metals, the color of the emission of vapors of which, when excited in the plasma of nanosecond discharges, corresponds to the color of high-altitude atmospheric discharges, were chosen as the material of the electrodes.	2	1	The topic is timely and the results provided are interesting.			nano12040652_perova	0
+The color of the glow of the discharge plasma at electrodes made of various metals is associated with excitation certain energy levels of particles in the vapors of these metals. So, when using electrodes made of aluminum, steel and copper, we observed the glow of red, blue and green colors, respectively. The different colors of the discharge when changing the material of the electrodes are determined not by the spark or arc stages, but by bright spots on the electrodes, which are formed due to the explosive emission of electrons [Mesyats, G.A. Ecton mechanism of the vacuum arc cathode spot. IEEE transactions on plasma science, 1995, 23(6), pp. 879-883. (DOI: 10.1109/27.476469)]. These areas in the photographs have a bright white color (see, for example, the photographs in Figures 3, 6, 7). In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material. High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap. However, emission of individual particles is determined by their temperature, it corresponds to the Planck radiation and is broadband. We note once again that in this work, to obtain metal vapors, as well as metal nano- and microparticles, a pulsed nanosecond discharge in a non-uniform electric field was used.	2	1	The paper provides interesting data concerning the colors of some specific kinds of plasma discharges in different gases and compares them to atmospheric discharges, considering the presence of metal vapors and nanoparticles coming from the electrodes.			nano12040652_perova	0
+Several references was added.	2	1	The article may be of interest, but for researchers in the field of plasma physics and gas discharges.			nano12040652_perova	0
+The main experiments were carried out with discharges in air, and the air pressure was chosen close to the pressures of high-altitude discharges. Other gases, such as argon, were chosen to better demonstrate the discoloration of the discharge.	2	1	2) The article is more descriptive in nature, there is no explanation of physical laws, but which are misleading readers.			nano12040652_perova	0
+The manuscript has been revised. This judgment was removed from the text of the manuscript.	2	1	Reply: In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence.			nano12040652_perova	0
+Thanks for the advice. We plan to do this in our next work.	2	1	This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc.			nano12040652_perova	0
+The second part of this analysis, titled: “Body composition assessment in Mexican children and adolescents. Part 2: Cross-validation of three bio-electrical impedance methods against dual X-ray absorptiometry for whole-body and regional body composition”, has already been accepted by Nutrients #1604578. The two analyses were conducted on the same children’s database and closely complement each other. As the accepted manuscript already includes the phrase ‘Part 2’ we prefer to keep ‘Part 1’ here ABSTRACT 2.	2	1	"I take from the results that those sentences were describing the Bland-Altman procedure, and if so, adding some text along the lines of ""In this procedure..."" would be helpful."			nu14051073_makarova	0
+Corrected, we adjusted this paragraph together with the next one to give a clearer explanation of the motivation of this study.	2	1	Part 2: Cross-validation of three bio-electrical impedance methods against dual X-ray absorptiometry for whole-body and regional body composition”, has already been accepted by Nutrients #1604578.			nu14051073_makarova	0
+We have adjusted accordingly. We hope now we can give a clearer explanation about the motivation of this study.	2	1	Table 1; Body Composition Variables: The mean values do not all agree with those in supplementary Table 1.			nu14051073_makarova	0
+Clarified in lines 164.	2	1	We have provided new evidence for the Mexican population that all the methods have utility for this purpose.			nu14051073_makarova	0
+The data used was total body with head, as recommended for the ISCD when using DXA for body composition instead of for bone densitometry. We clarified and referenced this in the text, lines 204-208.	2	1	06720, Mexico City, Mexico E-mail: desiree_15@hotmail.com, dradesireelopez@gmail.com Dr. Márquez 162, Col. Doctores, C.P.			nu14051073_makarova	0
+This was a typo, we corrected it. Line 260.	2	1	I am not familiar with the Bland-Altman method, so was unclear when I read the methods section whether the text in lines 204-207 was explaining the Bland-Altman method or describing a separate procedure.			nu14051073_makarova	0
+We corrected (lines 287-288).	2	1	None of the methods are ideal – all are flawed (in comparison with a gold standard).			nu14051073_makarova	0
+Indeed, it was like that. We have changed it to show only the data of the 288 subjects (without the 5 outliers).	2	1	None of the methods are ideal – all are flawed (in comparison with a gold standard).			nu14051073_makarova	0
+This was because of the inclusion of the 5 outliers in table 1, which has been corrected.	2	1	Furthermore, some of the biases for individual methods can be resolved by the publication of method-specific reference data, whereby all data can be converted to method-specific z-scores.			nu14051073_makarova	0
+We have	2	1	Please find our answers in the following lines.			nu14051073_makarova	0
+Added, the analysis was for the total sample and by subgroups by age and sex, line 287 14.	2	1	Added, the analysis was for the total sample and by subgroups by age and sex, line 287 P12 L283: Is Table 4 necessary?			nu14051073_makarova	0
+We have corrected as recommended. Table 4 sent to supplementary material as supplementary table 3.	2	1	P3 L125: DXA: Was the head ROI excluded from the analyses (as per recommendation by the ISCD)?			nu14051073_makarova	0
+Changed as supplementary figure 3.	2	1	My comments may reflect my ignorance, but I offer them nonetheless as other readers may also be confused by some sections as I was.			nu14051073_makarova	0
+Changed, line 19 INTRODUCTION 3.	2	1	We have changed it to show only the data of the 288 subjects (without the 5 outliers).			nu14051073_makarova	0
+We have stated in the conclusion that while individual methods show bias relative to the 4-component reference, the high correlations indicate that all the methods perform well in ranking individual children as having high or low FFM and fat mass. This ranking is itself very valuable in routine clinical care, particularly for longitudinal assessment. We have provided new evidence for the Mexican population that all the methods have utility for this purpose. Furthermore, some of the biases for individual methods can be resolved by the publication of method-specific reference data, whereby all data can be converted to method-specific z-scores. Publishing such reference data is a further aim of our project.	2	1	It may be worth considering moving Table 4 to the supplementary file and bringing supplementary Table 1 into the main document.			nu14051073_makarova	0
+added	2	1	We have found your recommendations and commentaries of great value and have made appropriate changes.			nu14051073_makarova	0
+Added to supplementary figure 1 and main figures 1 and 2 3.	2	1	)” Is there uncertainty of the number of participants or is this merely a typo?			nu14051073_makarova	0
+Specified in limitations lines 993-997. Important to say is that ~89% of Mexican population is Hispanic. Only 6.6% belong to indigenous population and 5.9% to Afro-Mexican ethnic groups.	2	1	)” Is there uncertainty of the number of participants or is this merely a typo?			nu14051073_makarova	0
+Corrected. In the previous version we put data of the whole sample of 293 subjects (including 5 outliers that should not been there). We have corrected the data, and now we only present data on the 288 subjects for both tables.	2	1	If not, DXA analysis should be redone and all relationships recalculated.			nu14051073_makarova	0
+Added to table 1.	2	1	Concluding that methods differ is not surprising.			nu14051073_makarova	0
+Thank you for your comment, we have tried to make the abstract less fragmented -Theoretical framework: it is very updated.	2	1	Journal of nutritional science and vitaminology, 61(Supplement), S66-S68.			nu14122489_perova	0
+Thanks for your comments, we have found a more updated reference (reference 18), cited in lines 96-98 to explain more the importance of HRQoL.	2	1	Diet is only expected to influence 2 of these 5 dimensions.			nu14122489_perova	0
+Line 501-503 We have added that we used a large sample size and that this was powerful enough to detect significant differences within sub-groups. We have also added a further description of our ability to capture results in a snapshot of time. I hope this clarifies this aspect.	2	1	Schools may be making the lunches to a budget rather than to a healthy diet criteria which is more expensive.			nu14122489_perova	0
+Thank you for your comment, the updated figures and tables are now at the end of the paper.	2	1	The basic information is not there in the paper: “KIDSCREEN-10 (KS-10) is derived from the KIDSCREEN-27, and provides a single index of global QoL using ten items related to physical well-being, psychological well-being, autonomy and parent relation, social support and peers, and school environment” See how these researchers have described the KIDSCREEN-10 .			nu14122489_perova	0
+Thank you for your comment, we are glad that you think so	2	1	Effects of socioeconomic status on nutrition in Asia and future nutrition policy studies.			nu14122489_perova	0
+Thank you for your comments about the instruments and the associated domains. We have updated these and aimed to explain them in more detail and with more clarity, sections 2.5, 2.6 and 2.7 lines 193-196, 215-225, 236-244.	2	1	The level of reporting of the core assessment instruments and their items, dimensions and subdomains needs to be enhanced.			nu14122489_perova	0
+These issues have been addressed. The tables and figures are more clear and more detailed. We have provided are reasoning for the choices of statistical methods.	2	1	Have given it minor but it is needs more in the tables			nu14122489_perova	0
+Thank you for this concern. We have aimed to explain that the KS-10 takes items from the longer KS versions but does not measure for each dimension as it is a general score. However, you have made a great point that some associations may be lost on the specific dimensions, we have tried to explain this reasoning such the burden of participation is lower to complete the KS-10 version lines 493-494 . We wanted to use the general score so that results can be compared with other studies and in other countries, which is one of the advantages of KS-10. Thank you for the insight, this has made us reflect on the KS-10 from the longer versions more.	2	1	2020 Jul;23(10):1754-1765. doi: See their Table 5.			nu14122489_perova	0
+We have now recognised the impact and importance of income to a greater depth and are thankful for your comments on this matter. It is a shame that we cannot say anything about income in this article as this was not measured, only parental education, however this has been suggested for future studies lines 535-537 .	2	1	The authors may know what these are, but many reader will not.			nu14122489_perova	0
+We have now made a recommendation to investigate this in more depth and to compare diet quality between school time and home time. It is important to assess if the lunches are the same across students of different SES, some insight on this is given in lines 522 and 534 and that an issue could be the diet quality provided at home as school lunches may even-out diet-related inequalities. Thank you for the recommendation as this is an important aspect of adolescents’ diet.	2	1	I am a not European reviewer and so found the RADDS a rather restricted list without some meat.			nu14122489_perova	0
+Thank you for your comments on this matter, we have edited this and hope it makes more sense now and that it does not make that assumption any longer, see lines 48-50.	2	1	The term is usually SES social economic status so it should be SES- F if it is social economic status – factors.			nu14122489_perova	0
+We understand your concern, there are a lot of abbreviations, the abbreviations have been re-written in each section to help follow the flow, and we have now added an abbreviation and key word table before the introduction to provide additional help for readers, thank you for this comment lines 37-45.	2	1	The authors may know what these are, but many reader will not.			nu14122489_perova	0
+Thank you for your great comment, we want the dimensions to be clear for the readers and this is an important observation. We have now edited how the SHEIA and RADDS variables are explained as well as the KS-10, sections 2.5 and 2.6 and we hope they are clearer to understand lines 193-196, 215-225, 236-244.	2	1	The American journal of clinical nutrition, 87(5), 1107-1117.			nu14122489_perova	0
+Thank you for your comments, it is important to mention status and the term SES-F has now been incorporated. We have also now used the term parental education and in section 2.7 we have mentioned that this was used as a proxy for SES, lines 257-258. This is to make it clearer that we only had access to parental education and no other SES-Fs and we hope that this is clearer now, we have used parental education instead of SES in lines 21, 111-114 and 149-150. We have also emphasised the importance of parental income, lines 419-428, 503-506, 533-537 and 550-555.	2	1	Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD.			nu14122489_perova	0
+Thank you, we agree that income as well as education plays a role in healthy food choices. However, the results in the article we are referring to looked at parental education, not income. As we did not have access to data in parental income, we were not able to include that variable in our paper	2	1	The first sentence assumes that poor diet is always associated with adolescents, this is incorrect, Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing			nu14122489_perova	0
+Although a higher education might lead to a higher income we unfortunately do not have the data to look at that but as previously mentioned we have now stressed the importance of parental income, lines 419-428, 503-506, 533-537 and 550-555, thank you for pointing this out.	2	1	Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			nu14122489_perova	0
+Thank you for your comment, income is most certainly important and we have now emphasised its importance in lines 419-428, 503-506, 533-537 and 550-555. We also have mentioned that parental education is being used as a proxy for SES lines 257-258 but we cannot infer anything about income as this was not measured, only parental education, not overall SES.	2	1	Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet.			nu14122489_perova	0
+Thank you for suggesting that we reference these two papers, Darmon and Drewnoski was very insightful and most certainly useful for this article, and we now make it clearer that education is a factor of SES and a proxy for income. Lines 419-428 offer an insight into income and affluence, however we have elaborated more on income in future perspectives 533-537 and in the conclusion, lines 550-555 as it may, as you say, play a significant role in these associations.	2	1	To assist the reader’s comprehension, explain the instruments more as you develop the paper.			nu14122489_perova	0
+Thank you for your comment, we want to make the definition of this index as clear as possible and we have edited this in section 2.5.1 and included the sub-components, lines 193-196. We have also cited that reference in both sections 2.5.1 and 2.5.2 so that readers can find a more detailed description. We have also edited section 2.5.2, lines 215-225 so that the RADDS index is also easier to understand and have mentioned some of the sub-components.	2	1	Journal of nutritional science and vitaminology, 61(Supplement), S66-S68.			nu14122489_perova	0
+Thanks for your observation, we have now tried to describe the KS-10 with more details, and to make it clearer, section 2.6, 236-244.	2	1	Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD.			nu14122489_perova	0
+We are very pleased that you found this interesting!	2	1	Diet is not expected to have any influence of parent relations, social relations or peers but your study may find an influence on psychological well-being and even school environment.			nu14122489_perova	0
+Thank you for this comment, the figure may have been minimised and red meat and poultry are now visible which may not have been before, apologies for this, also we have mentioned some of the sub-components in lines 215-225. Also the figures and tables became distorted when the manuscript was uploaded, we have fixed this.	2	1	Journal of nutritional science and vitaminology, 61(Supplement), S66-S68.			nu14122489_perova	0
+Thank you for your comment. However, we think chi-squared is the appropriate statistical test as we are only comparing the proportions of distribution between categorical variables, we are not comparing the means between more than two groups which is what ANOVA is used for. We have not used chi square to assess analysis of variance, we have now made it clearer that frequency distribution is being assessed in the statistical methods section In terms of gender and education: the mean, standard deviation, df and sig t or F test need to be reported in the tables.	2	1	This is the core of your study: does diet have an influence on psychological wellbeing?			nu14122489_perova	0
+Thanks for this comment, we have incorporated t and df scores, see tables 1 and 4.	2	1	If income is not important than the paper should be the on home education level and home diet not home SES and home diet.			nu14122489_perova	0
+This has been put in the appendix, table A1, page 18, thanks for the suggestion The regression analyses “p” value is reported, but the beta values and significance must also be reported.	2	1	The first sentence assumes that poor diet is always associated with adolescents, this is incorrect, Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing			nu14122489_perova	0
+Great that you mentioned the beta values, the coefficients in the tables are in fact unstandardised beta coefficient values, this has been made clearer in the tables. The tables have diet (ind variable) on the left and the dependent variable KS-10 is on the top to indicate that interaction, then this is stratified by gender. I hope this makes more sense now.	2	1	The level of reporting of the core assessment instruments and their items, dimensions and subdomains needs to be enhanced.			nu14122489_perova	0
+We had a problem with uploading the figures and they became distorted, thank you for your comments, we have fixed this and incorporated t and df values.	2	1	It needs minor revision in places but the results need more attention.			nu14122489_perova	0
+We appreciate your concern regarding the KS-10 item domains. The reason we chose to use the general item value is that it is less burdensome than the longer versions and it is best practice to use the general score and not to directly try to analyse for each dimension. We are not sure that it is a fact that diet can not influence parent relations or social relations, if diet can help to improve quality of life then this may improve how an adolescent feels and their emotions which ultimately may have an influence on relations to others. However, we realise that we may miss out other findings and have mentioned this as a limitation, lines 496-497 and we appreciate your views on this matter. We have also incorporated more information about KS-10 reliability in measuring HRQoL, 236-244This is also a cross sectional study and so only associations can be established not causations.	2	1	Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD.			nu14122489_perova	0
+Great comments, we have elaborated more on income, lines 419-428, 503-506, 533-537 and 550-555. We have also included a detailed section about the significance of school lunches and education in Sweden across different SES groups lines 528-532. In Sweden the school lunch is of rather high diet quality, reaching many of the national dietary recommendations and is provided free of charge regardless of income or SES. However, it would be insightful to a complete a deeper analysis investigating the differences of school quality across differing socio-demographic areas. Nonetheless, the problem related to diet quality may lie in the food provided at home as financial constraints are most likely to be of more significance, lines 528-534.	2	1	The important issue is in the method section the reader needs understand the dimensions and sub-dimension that make up within each survey.			nu14122489_perova	0
+No comments to address (‘Yes’ ticked for all assessment criteria). We thank Reviewer 1 for their encouraging feedback.	2	1	Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).			obesities2010010_makarova	0
+Thank you for your encouraging feedback on the potential reach of the findings. We have added at statement to this effect in the Discussion: Although based on a restricted area of exploration, Australian print news media only, our findings may have much broader significance for worldwide social trends and prompt the need for ongoing analysis of media reporting of obesity and weight-related public health policy. Future research could also extend our word embedding analysis to policy texts themselves, to draw direct correlations between media and policy data sources.	2	1	The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach.			obesities2010010_makarova	0
+We agree that analysis of associations between weight-biased language in news media and the prevalence of eating disorders and other mental disorders correlated with weight stigma would be informative to the weight stigma literature, however our input data for the natural language processing (NLP) were textual data only and do not take into account other data modalities such as images and/or clinical data (floating numbers). In other words, NLP is a text-analytical tool to understand the nuances of human language about a certain topic (obesity in our case). This is done by capturing the contextual relationships between words and sentences in text corpus. Furthermore, this suggestion goes beyond the scope of the present paper, which focuses on associations between language biases tied to individual and structural dimensions of obesity and changes in public health policy rather than associations between language biases and changes in mental disorder prevalence. An alternative approach with the aid of NLP, would be to add mental disorders as a dimension in the analyses, but this would require a comprehensive literature review to make sure the mental disorder keywords, and their dichotomous mappings, were inclusive. Given the turnaround time for the revision (10 days), unfortunately we cannot extend the analysis in this way, but we absolutely agree with Reviewer 2 that this is an important and interesting direction for future research that can be achieved with further application of the techniques we have developed for this paper. We have acknowledged investigation of the association between news reporting on obesity and mental disorders as a fruitful direction for future research in the Discussion as follows: It is also important to examine relationships between news media reporting of obesity and health outcomes over time given, for example, medium to large meta-analytic associations between weight stigma and mental health disorders such as anxiety, depression, eating disorders and other psychopathological symptoms [new citation to be added and numbered accordingly – see below]. Emmer, C., Bosnjak, M., Mata, J. The association between weight stigma and mental health: A meta-analysis. Obesity Reviews 2020, 21:e12935. doi:10.1111/obr.12935 Comment 3:	2	1	The manuscript reports a study about weight-biased language in the Australian media across the last decades.			obesities2010010_makarova	0
+We have provided new versions of the figure as a separate file to support editing/reproduction.	2	1	Is there any possible limit with this approach in the selection of the papers?			obesities2010010_makarova	0
+The research questions have been moved to the Introduction.	2	1	Author Response Reviewer 2: General comments: The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			obesities2010010_makarova	0
+This text was intended as an overview of what follows in the Results. We have moved it to the beginning of the Results and rephrased it as follows: In this section, we show the associations between obesity-related terms and the gender, healthiness, social status, and stereotype dimensions. These associations are subsequently cross-matched with the obesity policy timeline in the Discussion, to help interpret the context of change in biases over time.	2	1	I do not have particular concerns about the paper.			obesities2010010_makarova	0
+Figure 1 has been removed, along with the following associated text: Data extraction and analysis processes are illustrated in Figure 1.	2	1	The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			obesities2010010_makarova	0
+We have added the following text from this paper in the first paragraph of the discussion as it seems to fit best with our results: Such entrenched weight biases, persistent in the media, may lead to internalised or self-stigma among individuals with overweight and obesity that persist even after weight loss. A recent study [insert numbered citation] performed a semantic evaluation of body shapes in obesity surgery patients and overweight/obesity controls and found that both groups were more willing to accept positive adjectives as a match when BMI was low and negative adjectives as a match when BMI was high.	2	1	I think this aspect is interesting and linked to your results, showing a connection with clinical data.			obesities2010010_makarova	0
+We have now added a Limitations and Future Research section to the Discussion to address this point and others: There are two limitations in our data curation process, the automated approach we used to check and select papers. Firstly, automatic classifiers of any sort can include some irrelevant or false positive articles. Due to the large amount of articles in our dataset, in Step 2 of our methodology, we developed a machine learning binary classifier – a support vector machine - with 87.56% accuracy to automatically identify relevant articles (accuracy is the number of correct predictions made, divided by the total number of predictions made, and then multiplied by 100 to convert it into a percentage). As a rule of thumb, accuracy of a predictive model that is above 80% is very commonly used to summarise the performance of that model. Still, it doesn’t exclude the possibility of irrelevant articles in our final dataset, but that possibility is less than 12.44 percent and this has to be considered against the benefit of scale and efficiency that this method allows. We also acknowledge that our analysis limited is in not being able to consider visual content visual content, images within articles (known as image framing), which have been shown to carry stigmatising elements [48]. This is something that could be added to the approach by including image classification along with additional measures. Furthermore, even though the Dow Jones is one the largest news databases, it might still miss some articles or news sources (although this doesn’t relate to the automatic approach as such). This applies to social media, even though this would not span as long a timeframe.	2	1	Is there any possible limit with this approach in the selection of the papers?			obesities2010010_makarova	0
+No comments to address (‘Yes’ ticked for all assessment criteria). We thank Reviewer 1 for their encouraging feedback.	2	1	I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance.			obesities2010010_perova	0
+Thank you for your encouraging feedback on the potential reach of the findings. We have added at statement to this effect in the Discussion: Although based on a restricted area of exploration, Australian print news media only, our findings may have much broader significance for worldwide social trends and prompt the need for ongoing analysis of media reporting of obesity and weight-related public health policy. Future research could also extend our word embedding analysis to policy texts themselves, to draw direct correlations between media and policy data sources.	2	1	This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			obesities2010010_perova	0
+We agree that analysis of associations between weight-biased language in news media and the prevalence of eating disorders and other mental disorders correlated with weight stigma would be informative to the weight stigma literature, however our input data for the natural language processing (NLP) were textual data only and do not take into account other data modalities such as images and/or clinical data (floating numbers). In other words, NLP is a text-analytical tool to understand the nuances of human language about a certain topic (obesity in our case). This is done by capturing the contextual relationships between words and sentences in text corpus. Furthermore, this suggestion goes beyond the scope of the present paper, which focuses on associations between language biases tied to individual and structural dimensions of obesity and changes in public health policy rather than associations between language biases and changes in mental disorder prevalence. An alternative approach with the aid of NLP, would be to add mental disorders as a dimension in the analyses, but this would require a comprehensive literature review to make sure the mental disorder keywords, and their dichotomous mappings, were inclusive. Given the turnaround time for the revision (10 days), unfortunately we cannot extend the analysis in this way, but we absolutely agree with Reviewer 2 that this is an important and interesting direction for future research that can be achieved with further application of the techniques we have developed for this paper. We have acknowledged investigation of the association between news reporting on obesity and mental disorders as a fruitful direction for future research in the Discussion as follows: It is also important to examine relationships between news media reporting of obesity and health outcomes over time given, for example, medium to large meta-analytic associations between weight stigma and mental health disorders such as anxiety, depression, eating disorders and other psychopathological symptoms [new citation to be added and numbered accordingly – see below]. Emmer, C., Bosnjak, M., Mata, J. The association between weight stigma and mental health: A meta-analysis. Obesity Reviews 2020, 21:e12935. doi:10.1111/obr.12935 Comment 3:	2	1	This study deals with question that needs important discussion for obesity-related public health policy development.			obesities2010010_perova	0
+We have provided new versions of the figure as a separate file to support editing/reproduction.	2	1	The topic is interesting, and the manuscript is clear.			obesities2010010_perova	0
+The research questions have been moved to the Introduction.	2	1	This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			obesities2010010_perova	0
+This text was intended as an overview of what follows in the Results. We have moved it to the beginning of the Results and rephrased it as follows: In this section, we show the associations between obesity-related terms and the gender, healthiness, social status, and stereotype dimensions. These associations are subsequently cross-matched with the obesity policy timeline in the Discussion, to help interpret the context of change in biases over time.	2	1	I think this aspect is interesting and linked to your results, showing a connection with clinical data.			obesities2010010_perova	0
+Figure 1 has been removed, along with the following associated text: Data extraction and analysis processes are illustrated in Figure 1.	2	1	I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance.			obesities2010010_perova	0
+We have added the following text from this paper in the first paragraph of the discussion as it seems to fit best with our results: Such entrenched weight biases, persistent in the media, may lead to internalised or self-stigma among individuals with overweight and obesity that persist even after weight loss. A recent study [insert numbered citation] performed a semantic evaluation of body shapes in obesity surgery patients and overweight/obesity controls and found that both groups were more willing to accept positive adjectives as a match when BMI was low and negative adjectives as a match when BMI was high.	2	1	This study deals with question that needs important discussion for obesity-related public health policy development.			obesities2010010_perova	0
+We have now added a Limitations and Future Research section to the Discussion to address this point and others: There are two limitations in our data curation process, the automated approach we used to check and select papers. Firstly, automatic classifiers of any sort can include some irrelevant or false positive articles. Due to the large amount of articles in our dataset, in Step 2 of our methodology, we developed a machine learning binary classifier – a support vector machine - with 87.56% accuracy to automatically identify relevant articles (accuracy is the number of correct predictions made, divided by the total number of predictions made, and then multiplied by 100 to convert it into a percentage). As a rule of thumb, accuracy of a predictive model that is above 80% is very commonly used to summarise the performance of that model. Still, it doesn’t exclude the possibility of irrelevant articles in our final dataset, but that possibility is less than 12.44 percent and this has to be considered against the benefit of scale and efficiency that this method allows. We also acknowledge that our analysis limited is in not being able to consider visual content visual content, images within articles (known as image framing), which have been shown to carry stigmatising elements [48]. This is something that could be added to the approach by including image classification along with additional measures. Furthermore, even though the Dow Jones is one the largest news databases, it might still miss some articles or news sources (although this doesn’t relate to the automatic approach as such). This applies to social media, even though this would not span as long a timeframe.	2	1	I think this aspect is interesting and linked to your results, showing a connection with clinical data.			obesities2010010_perova	0
+Figures have been revised with addition of arrows and text to represent the data. Since a large number of ILs were screened, susceptible check (SC) was used with a set of ILs and hence SC couldn’t be shown in all figures. However, SC ‘HR12’ for blast disease was shown in all blast screening figures 3 to 5. Similarly, in Fig 5, ‘TN1’ and ‘Improved Samba Mahsuri’ as susceptible and resistant checks respectively for BB in comparison with IL-19031 were shown. Authors once again thank the reviewer for pointing out the mistake in legends. Now, we have rephrased the legends clearly describing all the terms.	2	1	The last paragraph of results is about background selection.			plants11050622_makarova	0
+Figure 1 legend has been revised with inclusion of the details of kharif and rabi seasons. kharif is the wet season with crop growing period from June to November and rabi is the dry season with crop growing period from December to May. We described kharif as wet season and rabi as dry season in the materials and methods section also.	2	1	The main issue with the ms is the quality of the figures: they are not clearly representing the data, arrows and text may help the reader, the controls are missing in most cases, and a general lack of precision is affecting them.			plants11050622_makarova	0
+Since the number of ILs is large, presenting phenotypic data for each IL will result in increasing the size of the main tables, hence data was earlier presented in supplementary tables. As suggested by the reviewers, we have revised the tables and presented the mean phenotypic data of BB, blast and drought screening in the main tables for each IL in parenthesis. Column on ‘no. of genes/QTL’ has been removed as suggested. Entry nos have been replaced with IL No as suggested.	2	1	With minor grammatical revisions, the manuscript can be accepted as is.			plants11050622_makarova	0
+The present work is not essentially a backcross breeding program aimed at development of near isogenic lines. However, Krishna Hamsa was the common background into which several genes/QTL were targeted from multiple donors and considering the morphological similarity between 27 ILs and Krishna Hamsa, background selection was done retrospectively. BGS validated our observations on morphological similarity. The same has been discussed in the 5th para under ‘discussion’. Also results on BGS have been presented under subsection 2.3 of results with data on polymorphic markers for BGS in supplementary table S8.	2	1	Indicate DFF = days to fifty percent flowering.			plants11050622_makarova	0
+“()” have been removed while mentioned the numbers of the ILs and sentences have been revised appropriately in the manuscript.	2	1	Dear Editor, Thank you for inviting me to review this manuscript.			plants11050622_makarova	0
+The above mentioned lines have been checked and found either spelling mistakes or revision of sentences. Accordingly, corrections were made.	2	1	How did you calculate those values on those comparison types?			plants11050622_makarova	0
+BLB has been replaced with BB throughout the manuscript.	2	1	It is also acknowledged that this paper is probably the first of many papers to emerge from the study.			plants11050622_makarova	0
+Legends of figures and tables and text in the results section have been revised with explanation of acronyms as suggested.	2	1	On Supplementary table S2, does significant mean p-value?			plants11050622_makarova	0
+The sentence here is required to maintain flow of the subsequent content.	2	1	The last paragraph of results is about background selection.			plants11050622_makarova	0
+Both lines explain our observations in different sets of ILs. Sentence at 351 explains susceptibility in ILs despite possessing the targeted gene/QTLs while sentence at 414 explains resistance in ILs despite the absence of targeted gene/QTLs.	2	1	of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion.			plants11050622_makarova	0
+The 85 ILs is a sum total of nine ILs marker positive to blast- R genes, nine ILs harbouring QTLs for drought tolerance and 67 ILs marker positive to BB-R genes mentioned at the beginning of 4th paragraph of discussion. As suggested, we have added in brief about the same at line 414.	2	1	The last paragraph of results is about background selection.			plants11050622_makarova	0
+Authors profusely thank the reviewer for the appreciation.	2	1	the 85 ILs presented at 414 seem to be a bit out of the blue: a short intro to where they are coming from would help the reader.			plants11050622_makarova	0
+Yes, we agree that there were some typo and spelling mistakes in the manuscript. The manuscript has been thoroughly revised for the same.	2	1	On Supplementary table S2, does significant mean p-value?			plants11050622_makarova	0
+Authors feel greatly encouraged and motivated with the reviewer’s comments.	2	1	For example on L203 “CD”, L211 “SES“, L228 “UBN“ and L332 “ICAR-IRRR”.			plants11050622_makarova	0
+‘=261%’ has been corrected to ‘+261%’ and typo error of ‘linkes’ corrected to ‘linked’	2	1	Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups.			plants11050622_makarova	0
+Thank you for the positive comments.	2	1	Author Response Point-by-point response to the reviewer's comments are given below The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			plants11050622_makarova	0
+We appreciate the suggestions from the reviewer, which has resulted in improving the message of the manuscript.	2	1	L562: the section of statistical analysis should add more details.			plants11050622_makarova	0
+Added references appropriately at two places as suggested by the reviewer.	2	1	L127: Please add the explanation of “boro season”.			plants11050622_makarova	0
+Explanation for boro season has been added.	2	1	Table and main text are independent, so the authors have to describe the table more carefully.			plants11050622_makarova	0
+Legends of the supplementary tables have been revised and inference of the table is given in foot note.	2	1	How did you calculate those values on those comparison types?			plants11050622_makarova	0
+Authors once again thank the reviewer for the valuable suggestion. More details on the statistical analysis have been added as suggested.	2	1	On Supplementary table S3, similar questions as S2, please also explain.			plants11050622_makarova	0
+Yes, significant means p-value. In the supplementary tables, expanded form of DFF is given as suggested. Treatments refers to introgression lines and check to control. For uniformity, we have changed the terminology to treatment instead of using IL or variety and check to control in all the revised supplementary tables and rephrased the legends accordingly.	2	1	Please add some gel pictures of the foreground selection markers you used in order to visualize the genotyping results and showed the polymorphism of these markers on gel.			plants11050622_makarova	0
+Similar to Supplementary Table S2, S3 has been revised	2	1	Also, the legends are not clearly describing all the terms and should be revised.			plants11050622_makarova	0
+CD is the critical difference at 1% and 5% level of significance (p-value) for testing of significant differences among the ILs. The details of CD calculation have been added in the materials and methods section as suggested.	2	1	The paper itself is well written, although 1) somewhat results are partially descriptive and partially inferential.			plants11050622_makarova	0
+Supplementary table S5 is on CD and corrections have been addressed as suggested as at S. No 7. Supplementary table S6 have been modified by shifting data of BB and blast scores to main table. Each PC group is mentioned on top as sub heading at the start of each group. Each IL is presented only once in the entire table under separate PC groups.	2	1	Explaining briefly which and what are the Indian seasons during which experiments have been conducted would help clarity.			plants11050622_makarova	0
+Full names of the abbreviations have been added as suggested.	2	1	Also, the legends are not clearly describing all the terms and should be revised.			plants11050622_makarova	0
+Subtitle has been added as suggested.	2	1	Then the table can be more informative and ease to read.			plants11050622_makarova	0
+A representative gel picture has been added as suggested.	2	1	On Supplementary table S3, similar questions as S2, please also explain.			plants11050622_makarova	0
+Table S8 represents polymorphic markers between pairs of parents including some common polymorphic markers. Hence the total number represented in the table S8 is not additive and not matching with the numbers given in the text. 124 is the total polymorphic markers excluding repetition. 27 ILs were selected based on their agro-morphological similarity with recurrent parent ‘Krishna Hamsa’ and evaluated for background recovery. The list of 27 ILs has been included in the revised manuscript as suggested.	2	1	Correct it It was a pleasure to read this manuscript.			plants11050622_makarova	0
+Mention of appendix at L602 is a typo error and has been removed	2	1	Author Response Point-by-point response to the reviewer's comments are given below The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			plants11050622_makarova	0
+We agree with your comments. The term ‘expression’ has, therefore, been changed to ‘abundance’ or ‘level’ throughout the manuscript.	2	1	The statistical significance of any result would be evaluated using multivariate analysis.			reprodmed3020015_makarova	0
+All experiments were repeated at least two times. We have failed the description regarding the reproducible results in the previously submitted manuscript, and so revised the section of the methods in the revised manuscript (Page 4, lins143-143).	2	1	This result must be demonstrated in a substantively convincing manner.			reprodmed3020015_makarova	0
+We agree that AG-205 is not a specific inhibitor of PGRMC1. Therefore, we examined the effects of siRNA-mediated PGRMC1 knockdown on decidualization. As you mentioned, this issue was not appropriately described in the previously submitted manuscript. The citations have been inserted in the revised manuscript (page 8, lines 258-263).	2	1	This result must be demonstrated in a substantively convincing manner.			reprodmed3020015_makarova	0
+The name of the reagent has been corrected to AG-205 consistently throughout the manuscript.	2	1	“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments.			reprodmed3020015_makarova	0
+The sentence has been separated according to your suggestion.	2	1	There was no control in PGRMC1 genomic/CRISPR KO cells (not si/miRNA KO) to demonstrate that effects induced by AG-205 were due to PGRMC1.			reprodmed3020015_makarova	0
+The description has been modified as followed (page 2, line 69). ‘Endometrial samples were obtained from Japanese patients with a normal menstrual cycle, ….’.	2	1	Such changes may occur at the translational or post-translational level.			reprodmed3020015_makarova	0
+The primary cultured ESCs we used were confirmed immunocytochemically to be positive for vimentin, a marker for stromal cells, and negative for cytokeratin, a marker for epithelial cells (please see above figure). ESCs at early passages (between 2 and 6 passages) were used for the experiment. In this study, the passaged cells were used without freezing. However, we have confirmed that primary ESCs can be cryopreserved in CELLBANKER, and retain characteristics such as responsiveness to decidual stimuli. These descriptions have been added to the revised manuscript (page 3, lines 96-99). Figure. Immunocytochemistry (ICC) of vimentin and cytokeratin in isolated human ESCs.	2	1	In primary endometrial stem cell (ESC) culture they demonstrate that chemical induction (by db-cAMP/P4) of pseudo-decidualization in culture is accompanied by decreased PGRMC1 protein levels (fig.2).			reprodmed3020015_makarova	0
+The protein samples were separated by SDS-PAGE (5-20% gradient gel) and transferred electrophoretically to polyvinylidene difluoride membranes (8 cm x 8.5 cm) for 60 min at constant current of 128 mA using a semi-dry transfer system (ATTO, Tokyo Japan). These conditions have been added to the Materials and Methods section (Pages 3 and 4, lines 127-130).	2	1	The authors have addressed most of my concerns.			reprodmed3020015_makarova	0
+The catalogue number of the antibodies (PI-1000, PI-2000) were inserted (Page 4, line 134).	2	1	The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed.			reprodmed3020015_makarova	0
+The immunoreactive bands were visualized using an enhanced chemiluminescence (Western Lightning, PerkinElmer, Inc., Waltham, MA, USA) and analyzed with ImageQuant LAS 500 (semi-auto mode; GE Healthcare Japan, Tokyo Japan) (Page 4, lines 136-137).	2	1	The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			reprodmed3020015_makarova	0
+The densitometry analysis was carried out using the “Gel Plot” plug-in of the Image J software. This plug-in has been described in the Materials and Methods (Page 4, line 139) 12)	2	1	Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy.			reprodmed3020015_makarova	0
+As you indicated, all data are expressed consistently as means and standard deviation (Figs 1 to 4). In our previously submitted manuscript, we had described carelessly the data of as for figure 5B and C with s.d.	2	1	The work would be worthwhile publishing in a revised format, where it should be a well cited publication.			reprodmed3020015_makarova	0
+Thank you for pointing this out. The mistakes have been corrected (Pages 4,5 and 7, lines 160, 174 and 217).	2	1	Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.			reprodmed3020015_makarova	0
+Based upon your suggestions, additional experiments have been performed and added individual Western blotting data from there different ESCs (named #1-3) in Figure 2.	2	1	PGRMC1 protein instability could be the dominant effect involved, with miRNA transcript regulation playing only a minor part.			reprodmed3020015_makarova	0
+Western blotting for PGRMC1 in figures 2, 3, and 4 have been replaced in the data with the whole molecular weight range. As you pointed out, PGRMC1 is known to be modified by sumoylation or ubiquitinylation which results in high molecular weight. The right panel shows the western blotting of PGRMC1. There are some higher molecular bands (white arrowhead) in lysates of ESCs, however, the band’s intensity was not changed by PGRMC1 siRNA treatment. Therefore, we believe that these high molecular bands detected in our experiment were non-specific bands.	2	1	Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true).			reprodmed3020015_makarova	0
+As you pointed out, the possible involvement of posttranslational modification of endometrial PGRMC1 such as sumoylation and ubiquitinylation is interesting. Discussion regarding this possibility has been inserted in the revised manuscript (Page 9, lines 300-307). We will further examine whether posttranslational modification of PGRMC1 could be associated with decidualization in the next study.	2	1	Unfortunately, the authors have been unable to demonstrate the direct involvement of PGRMC1 downregulation in decidualization.			reprodmed3020015_makarova	0
+The mistake has been corrected.	2	1	There is some concern about the specificity of the AG-205 inhibitor here and throughout, which weakens the deductive reasoning and is discussed below.			reprodmed3020015_makarova	0
+All data of cell culture represent the mean ± SD of three independent experiments with at least repeated two times each experiment. The explanation of the experiments has been inserted the method (page 4, lines142-143).	2	1	PGRMC1 levels are also controlled by proteolysis, mediated by the ubiquitin proteasome system as described above.			reprodmed3020015_makarova	0
+Thank you for your valuable advice to prove the significance of PGRMC1 downregulation in decidualization. Salsano et al. (19), reported that overexpression of PGRMC1 in ESCs abrogated the decidual markers expression (page 8. Lines 253-254). We will try to examine the effects of enforced expression of codon-altered PGRMC1 which is resistant for siRNA on decidualization in next study.	2	1	Author Response Responses to Reviewer 1 We appreciate your review of our manuscript and constructive comments and suggestions.			reprodmed3020015_makarova	0
+Based upon your suggestions, we have added the data of PGRMC1 knockdown in figure 4. We confirmed efficient siRNA-mediate PGRMC1 knockdown by western blotting.	2	1	That new experimental result is what elevated the required changes to a major revision.			reprodmed3020015_makarova	0
+The statistical analysis was not appropriate in the former manuscript. The data has been re-evaluated by a two-way ANOVA followed by a Turkey-Kramer multiple comparison test (page 4, line13).	2	1	Author Response Responses to Reviewer 2 We appreciate your review of our manuscript and constructive comments and suggestions.			reprodmed3020015_makarova	0
+As you suggested, we should have presented the data in the same graph. However, the two experiments were performed independently. Therefore, we were not able to combine the results in the graph. Because treatment with P4 alone could not induce decidualization in the experiments, we examined the effect of PGRMC1 knockdown and inhibition on db-cAMP-induced decidualization. Further study would be needed to determine whether PGRMC1 is involved in the P4 action in endometrium.	2	1	The paragraph describes a result showing that the results are independent of P4.			reprodmed3020015_makarova	0
+The description has been corrected and cited the references in the section (page 7, lines 223-224).	2	1	Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) Lines 271-76.			reprodmed3020015_makarova	0
+New discussion regarding the need for PGRMC1 knockout studies to clarify the role of PGRMC1 in decidualization has been inserted in page 9,	2	1	Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?).			reprodmed3020015_makarova	0
+The error has been corrected.	2	1	If further mechanism exploration is carried out, the research will be more complete and persuasive.			reprodmed3020015_makarova	0
+Your correction is what we meant to mention. We have replaced this sentence in the revised manuscript. (page9, line 284-286) 27)	2	1	It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1.			reprodmed3020015_makarova	0
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, line 295-299).	2	1	The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required.			reprodmed3020015_makarova	0
+The report that PGRMC1 mediates H2O2-induced cell death in MCF7 cells has been inserted in discussion (page 10, lines 314-317)	2	1	This result must be demonstrated in a substantively convincing manner.			reprodmed3020015_makarova	0
+We have suggested the possibility that PGRMC1 downregulation may be involved in ESCs senescence during the decidualization (page 10, lines 319-321).	2	1	PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988).			reprodmed3020015_makarova	0
+As you pointed out, we infer that downregulation of PGRMC1 may be necessary for decidualization to proceed. To clarify this point, we further attempted to overexpress PGRMC1 by introducing expression vectors into primary ESCs under various conditions using lipofection and calcium phosphate transfection methods, as you suggested. Unfortunately, for reasons unknown, we were unable to establish ESCs overexpressing PGRMC1. In the present study, we found that not only knockdown of PGRMC1 but also functional inhibition with inhibitors promotes ESC decidualization in vitro. Furthermore, in a previous report, overexpression of PGRMC1 suppressed ESCs differentiation (Ref. 19). These results indicate that decreased PGRMC1 in the endometrium may be associated with accelerated dedifferentiation. As you suggested, we have changed the title significantly because further evidence was not available. We plan to establish a model of miR-resistant PGRMC1 overexpression using several ESCs cell lines to investigate the role of PGRMC1 in decidualization, and if possible, we would be happy to make this our next research topic.	4	1	The authors have addressed most of my concerns.			reprodmed3020015_makarova	0
+Thank you for correcting the sentence. The description has been corrected in accordance with your suggestion (page 4, lines 141-142).	4	1	This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			reprodmed3020015_makarova	0
+The error has been corrected (page 8, line 257).	4	1	PGRMC1 is known to be Sumoylated or ubiquitinylated and then degraded by the proteasome pathway, which lead to higher molecular weight species (PMID: 31067491).			reprodmed3020015_makarova	0
+The description has been modified to the following (page 8, lines 261-266). “Although we cannot exclude the possible non-specific action of AG-205 on ESCs, the effect of AG-205 treatment and PGRMC1 knockdown promoted in vitro decidualization. These findings suggested that PGRMC1 downregulation may promote ESC decidualization during the secretory phase.	4	1	Line 261. mechanisms HAVE not (plural mechanisms) Lines 261-262.			reprodmed3020015_makarova	0
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, lines 280-281).	4	1	In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			reprodmed3020015_makarova	0
+As you pointed out, we did not explain the relationship between PGRMC1 stability and post-transcriptional regulation of the protein. We have added the following description (page 9, lines 304-308). ”These reports suggested that PGRMC1 protein levels can be modulated by changes in the rates of transcription, translation, and degradation. Therefore, further studies would be needed to clarify the possible involvement of the post-translational modification and degradation that may regulate PGRMC1 protein stability in ESCs during decidualization.”	4	1	In response the authors have made minimal changes to the manuscript, effectively restricted to rewording the title, and several minor text changes.			reprodmed3020015_makarova	0
+We appreciate your review of our revised manuscript, constructive comments, and suggestions for improving the manuscript. We deeply understand the significance of the experiments using miR-98-resistant PGRMC stably expressing ESCs to directly prove the implication of the miR-mediated PGRMC1 regulation in decidualization. We are transfecting the miR-resistant PGRMC1 expression vector into ESCs cell lines to explore the role of PGRMC1 in decidualization but is not going well. As you advised, lentiviral transduction of the PGRMC1 along with fluorescent protein may enable us to effectively select the overexpressing cells without losing differentiation status in response to decidual stimuli. We have acknowledged this and suggested it as a topic for further research in the “Limitation” section of the revised manuscript following editor’s comment (page 10, lines 336-348). “Limitations: In this study, we proposed that miR-98-mediated PGRMC1 downregulation may be involved in decidualization, but were unable to examine the effects of miR-98-resistant PGRMC1 on differentiation due to technical difficulties establishing PGRMC1-overexpressing ESCs. miR-98 may not be the only miRNA that modulates PGRMC1 expression; therefore, comprehensive analysis of transcriptional, post-transcriptional, and post-translational regulation of endometrial PGRMC1 during decidualization is required. In addition, PGRMC1 can bind to various proteins, including receptors for epidermal growth factor [46] and insulin [47], cytochrome P450 [48], and serpine mRNA-binding protein 1 (SERBP1) [19]. Further, PGRMC1 may interact with proteins associated with endomembrane trafficking/cytoskeleton and mitochondrial functions in decidualizing ESCs [49]. Thus, we plan to investigate the modulation of endometrial PGRMC1 expression and the interaction between PGRMC1 and intracellular proteins during decidualization in the future.”	6	1	This requires stronger evidence than that currently presented.			reprodmed3020015_makarova	0
+Our responses to your comments are as follows. We appreciate your review of our revised manuscript, constructive comments, and suggestions. As you pointed out, we have deleted the proposed model of the study (Figure 6). We have added the “Strength and limitation of this study” and “Conclusions” in the revised manuscript in accordance with the academic editor’s comments.	8	1	This result must be demonstrated in a substantively convincing manner.			reprodmed3020015_makarova	0
+As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	In general, the manuscript refers often to PGRMC1 expression or downregulation.			reprodmed3020015_makarova	0
+Because of the ethical difficulties of isolating ESCs from disease-free healthy women, we routinely used ESCs isolated from the patients in surgical cases. ESC lines are commercially available (T-HESC, ATCC) and maybe useful tool to investigate the role of PGRMC1 in decidualization, as you pointed out. However, the cell lines are also established from the endometrium of leiomyoma and immortalized by transfection with hTERT. Therefore, we believe that ESCs used in this study are no different than the above cells and cell lines in examining the role of PGRMC1.In the future, we plan to use several ESCs and their cell lines to clarify the precise mechanisms of PGRMC1 downregulation and decidualization.	2	1	It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1.			reprodmed3020015_makarova	0
+We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript. We are thankful for the time and energy you expended.	4	1	The paragraph describes a result showing that the results are independent of P4.			reprodmed3020015_makarova	0
+The author provided explanations for the questions. The manuscript has been modified accordingly. I think the manuscript could be accepted now.	3	2	Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation.			reprodmed3020015_makarova	0
+We agree with your comments. The term ‘expression’ has, therefore, been changed to ‘abundance’ or ‘level’ throughout the manuscript.	2	1	This cell culture experiment also seems to be based upon just one primary cell culture, split into replicates.			reprodmed3020015_perova	0
+All experiments were repeated at least two times. We have failed the description regarding the reproducible results in the previously submitted manuscript, and so revised the section of the methods in the revised manuscript (Page 4, lins143-143).	2	1	However, in the first instance the manuscript cannot be published as it stands, and should therefore be rejected now with the stated option of subsequent resubmission only if new evidence is obtained.			reprodmed3020015_perova	0
+We agree that AG-205 is not a specific inhibitor of PGRMC1. Therefore, we examined the effects of siRNA-mediated PGRMC1 knockdown on decidualization. As you mentioned, this issue was not appropriately described in the previously submitted manuscript. The citations have been inserted in the revised manuscript (page 8, lines 258-263).	2	1	Cell culture methods must be much more clearly explained so that the reader can reconstruct each exact experiment.			reprodmed3020015_perova	0
+The name of the reagent has been corrected to AG-205 consistently throughout the manuscript.	2	1	Since this is the main publishable finding, the results as they stand do not merit publication.			reprodmed3020015_perova	0
+The sentence has been separated according to your suggestion.	2	1	This result must be demonstrated in a substantively convincing manner.			reprodmed3020015_perova	0
+The description has been modified as followed (page 2, line 69). ‘Endometrial samples were obtained from Japanese patients with a normal menstrual cycle, ….’.	2	1	The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.			reprodmed3020015_perova	0
+The primary cultured ESCs we used were confirmed immunocytochemically to be positive for vimentin, a marker for stromal cells, and negative for cytokeratin, a marker for epithelial cells (please see above figure). ESCs at early passages (between 2 and 6 passages) were used for the experiment. In this study, the passaged cells were used without freezing. However, we have confirmed that primary ESCs can be cryopreserved in CELLBANKER, and retain characteristics such as responsiveness to decidual stimuli. These descriptions have been added to the revised manuscript (page 3, lines 96-99).	2	1	The result must be shown for at least each ESC cell culture (each culture or patient named separately) obtained from each patient to demonstrate that the described results are representative of multiple patients, and not potentially atypical results from an atypical patient.			reprodmed3020015_perova	0
+The protein samples were separated by SDS-PAGE (5-20% gradient gel) and transferred electrophoretically to polyvinylidene difluoride membranes (8 cm x 8.5 cm) for 60 min at constant current of 128 mA using a semi-dry transfer system (ATTO, Tokyo Japan). These conditions have been added to the Materials and Methods section (Pages 3 and 4, lines 127-130).	2	1	And please provide basic information of the patient.			reprodmed3020015_perova	0
+The catalogue number of the antibodies (PI-1000, PI-2000) were inserted (Page 4, line 134).	2	1	This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues.			reprodmed3020015_perova	0
+The immunoreactive bands were visualized using an enhanced chemiluminescence (Western Lightning, PerkinElmer, Inc., Waltham, MA, USA) and analyzed with ImageQuant LAS 500 (semi-auto mode; GE Healthcare Japan, Tokyo Japan) (Page 4, lines 136-137).	2	1	Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool.			reprodmed3020015_perova	0
+The densitometry analysis was carried out using the “Gel Plot” plug-in of the Image J software. This plug-in has been described in the Materials and Methods (Page 4, line 139)	2	1	Therefore, a resubmission after major revision is recommended.			reprodmed3020015_perova	0
+As you indicated, all data are expressed consistently as means and standard deviation (Figs 1 to 4). In our previously submitted manuscript, we had described carelessly the data of as for figure 5B and C with s.d.	2	1	The authors mention technical difficulties in establishing ESCs that express miRNA-resistant PGRMC1.			reprodmed3020015_perova	0
+Thank you for pointing this out. The mistakes have been corrected (Pages 4,5 and 7, lines 160, 174 and 217).	2	1	“In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded.			reprodmed3020015_perova	0
+Based upon your suggestions, additional experiments have been performed and added individual Western blotting data from there different ESCs (named #1-3) in Figure 2.	2	1	The authors are strongly advised to change from e.g.			reprodmed3020015_perova	0
+Western blotting for PGRMC1 in figures 2, 3, and 4 have been replaced in the data with the whole molecular weight range. As you pointed out, PGRMC1 is known to be modified by sumoylation or ubiquitinylation which results in high molecular weight. The right panel shows the western blotting of PGRMC1. There are some higher molecular bands (white arrowhead) in lysates of ESCs, however, the band’s intensity was not changed by PGRMC1 siRNA treatment. Therefore, we believe that these high molecular bands detected in our experiment were non-specific bands.	2	1	This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA.			reprodmed3020015_perova	0
+As you pointed out, the possible involvement of posttranslational modification of endometrial PGRMC1 such as sumoylation and ubiquitinylation is interesting. Discussion regarding this possibility has been inserted in the revised manuscript (Page 9, lines 300-307). We will further examine whether posttranslational modification of PGRMC1 could be associated with decidualization in the next study.	2	1	In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			reprodmed3020015_perova	0
+The mistake has been corrected.	2	1	Our responses to your comments are as follows.			reprodmed3020015_perova	0
+All data of cell culture represent the mean ± SD of three independent experiments with at least repeated two times each experiment. The explanation of the experiments has been inserted the method (page 4, lines142-143).	2	1	The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation.			reprodmed3020015_perova	0
+Thank you for your valuable advice to prove the significance of PGRMC1 downregulation in decidualization. Salsano et al. (19), reported that overexpression of PGRMC1 in ESCs abrogated the decidual markers expression (page 8. Lines 253-254). We will try to examine the effects of enforced expression of codon-altered PGRMC1 which is resistant for siRNA on decidualization in next study.	2	1	Some of the tissue samples are from patients with endometriosis, then according to eutopic endometrium determinism, whether such selection will cause the research results to be inaccurate？ The sample size is small.			reprodmed3020015_perova	0
+Based upon your suggestions, we have added the data of PGRMC1 knockdown in figure 4. We confirmed efficient siRNA-mediate PGRMC1 knockdown by western blotting.	2	1	“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments.			reprodmed3020015_perova	0
+The statistical analysis was not appropriate in the former manuscript. The data has been re-evaluated by a two-way ANOVA followed by a Turkey-Kramer multiple comparison test (page 4, line13).	2	1	Result errors should not be expressed as SEM, but as standard deviation (s.d.).			reprodmed3020015_perova	0
+As you suggested, we should have presented the data in the same graph. However, the two experiments were performed independently. Therefore, we were not able to combine the results in the graph. Because treatment with P4 alone could not induce decidualization in the experiments, we examined the effect of PGRMC1 knockdown and inhibition on db-cAMP-induced decidualization. Further study would be needed to determine whether PGRMC1 is involved in the P4 action in endometrium.	2	1	The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).			reprodmed3020015_perova	0
+The description has been corrected and cited the references in the section (page 7, lines 223-224).	2	1	The discussion must qualify the results of Salsano et al.			reprodmed3020015_perova	0
+New discussion regarding the need for PGRMC1 knockout studies to clarify the role of PGRMC1 in decidualization has been inserted in page 9, lines 281-282.	2	1	The authors are strongly advised to change from e.g.			reprodmed3020015_perova	0
+The error has been corrected.	2	1	However, the control structure of comparisons is incorrect.			reprodmed3020015_perova	0
+Your correction is what we meant to mention. We have replaced this sentence in the revised manuscript. (page9, line 284-286)	2	1	I requested a major revision, involving expression of an miRNA-resistant PGRMC1 ORF to convincingly demonstrate the mechanistic involvement of PGRMC1 down-regulation in decidualization, rather than a temporal correlation.			reprodmed3020015_perova	0
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, line 295-299).	2	1	Enunciate what kind of role(s) PGRMC1 may be involved in (up/down regulation, when, how?).			reprodmed3020015_perova	0
+The report that PGRMC1 mediates H2O2-induced cell death in MCF7 cells has been inserted in discussion (page 10, lines 314-317)	2	1	Nine PGRMC1 lysines are known to be ubiquitinated (https://www.phosphosite.org/proteinAction.action?id=5744&showAllSites=true).			reprodmed3020015_perova	0
+We have suggested the possibility that PGRMC1 downregulation may be involved in ESCs senescence during the decidualization (page 10, lines 319-321).	2	1	Statistical sample sizes were unfavourably small.			reprodmed3020015_perova	0
+We appreciate your review of our revised manuscript, constructive comments, and suggestions for improving the manuscript. We deeply understand the significance of the experiments using miR-98-resistant PGRMC stably expressing ESCs to directly prove the implication of the miR-mediated PGRMC1 regulation in decidualization. We are transfecting the miR-resistant PGRMC1 expression vector into ESCs cell lines to explore the role of PGRMC1 in decidualization but is not going well. As you advised, lentiviral transduction of the PGRMC1 along with fluorescent protein may enable us to effectively select the overexpressing cells without losing differentiation status in response to decidual stimuli. We have acknowledged this and suggested it as a topic for further research in the “Limitation” section of the revised manuscript following editor’s comment (page 10, lines 336-348). “Limitations: In this study, we proposed that miR-98-mediated PGRMC1 downregulation may be involved in decidualization, but were unable to examine the effects of miR-98-resistant PGRMC1 on differentiation due to technical difficulties establishing PGRMC1-overexpressing ESCs. miR-98 may not be the only miRNA that modulates PGRMC1 expression; therefore, comprehensive analysis of transcriptional, post-transcriptional, and post-translational regulation of endometrial PGRMC1 during decidualization is required. In addition, PGRMC1 can bind to various proteins, including receptors for epidermal growth factor [46] and insulin [47], cytochrome P450 [48], and serpine mRNA-binding protein 1 (SERBP1) [19]. Further, PGRMC1 may interact with proteins associated with endomembrane trafficking/cytoskeleton and mitochondrial functions in decidualizing ESCs [49]. Thus, we plan to investigate the modulation of endometrial PGRMC1 expression and the interaction between PGRMC1 and intracellular proteins during decidualization in the future.”	4	1	What percentage acrylamide were the SDS PAGE gels.			reprodmed3020015_perova	0
+Our responses to your comments are as follows. We appreciate your review of our revised manuscript, constructive comments, and suggestions. As you pointed out, we have deleted the proposed model of the study (Figure 6). We have added the “Strength and limitation of this study” and “Conclusions” in the revised manuscript in accordance with the academic editor’s comments.	6	1	“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments.			reprodmed3020015_perova	0
+As you mentioned, our description of ESCs isolation may be inappropriate in terms of the patients with genetical differences. ESCs were isolated from histologically normal region in the patients with leiomyoma (age 42-45). It has been reported that ESCs obtained from the eutopic endometrium of the patients with endometriosis showed impaired decidualization. Therefore, we should consider the possible influence of leiomyoma on the decidual response, but we confirmed that the isolated ESCs have differentiated into decidual cells in response to cAMP analogue and progesterone, as you pointed out. The information of the patients has been inserted in the method 2)	2	1	And please provide basic information of the patient.			reprodmed3020015_perova	0
+Because of the ethical difficulties of isolating ESCs from disease-free healthy women, we routinely used ESCs isolated from the patients in surgical cases. ESC lines are commercially available (T-HESC, ATCC) and maybe useful tool to investigate the role of PGRMC1 in decidualization, as you pointed out. However, the cell lines are also established from the endometrium of leiomyoma and immortalized by transfection with hTERT. Therefore, we believe that ESCs used in this study are no different than the above cells and cell lines in examining the role of PGRMC1.In the future, we plan to use several ESCs and their cell lines to clarify the precise mechanisms of PGRMC1 downregulation and decidualization.	2	1	All reported results were observed in at least two independent experiments.			reprodmed3020015_perova	0
+As you pointed out, we infer that downregulation of PGRMC1 may be necessary for decidualization to proceed. To clarify this point, we further attempted to overexpress PGRMC1 by introducing expression vectors into primary ESCs under various conditions using lipofection and calcium phosphate transfection methods, as you suggested. Unfortunately, for reasons unknown, we were unable to establish ESCs overexpressing PGRMC1. In the present study, we found that not only knockdown of PGRMC1 but also functional inhibition with inhibitors promotes ESC decidualization in vitro. Furthermore, in a previous report, overexpression of PGRMC1 suppressed ESCs differentiation (Ref. 19). These results indicate that decreased PGRMC1 in the endometrium may be associated with accelerated dedifferentiation. As you suggested, we have changed the title significantly because further evidence was not available. We plan to establish a model of miR-resistant PGRMC1 overexpression using several ESCs cell lines to investigate the role of PGRMC1 in decidualization, and if possible, we would be happy to make this our next research topic.	2	1	The authors could consider whether an inducible miRNA-resistant PGRMC1 promoter could be designed.			reprodmed3020015_perova	0
+Thank you for correcting the sentence. The description has been corrected in accordance with your suggestion (page 4, lines 141-142).	2	1	Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.			reprodmed3020015_perova	0
+The error has been corrected (page 8, line 257).	2	1	It is important to determine whether these mechanisms are both operating.			reprodmed3020015_perova	0
+The description has been modified to the following (page 8, lines 261-266). “Although we cannot exclude the possible non-specific action of AG-205 on ESCs, the effect of AG-205 treatment and PGRMC1 knockdown promoted in vitro decidualization. These findings suggested that PGRMC1 downregulation may promote ESC decidualization during the secretory phase.	2	1	The authors have addressed most of my concerns.			reprodmed3020015_perova	0
+As noted, the sentence has been separated and corrected in accordance with your suggestion (page 9, lines 280-281).	2	1	The author provided explanations for the questions.			reprodmed3020015_perova	0
+As you pointed out, we did not explain the relationship between PGRMC1 stability and post-transcriptional regulation of the protein. We have added the following description (page 9, lines 304-308). ”These reports suggested that PGRMC1 protein levels can be modulated by changes in the rates of transcription, translation, and degradation. Therefore, further studies would be needed to clarify the possible involvement of the post-translational modification and degradation that may regulate PGRMC1 protein stability in ESCs during decidualization.”	2	1	In response the authors have made minimal changes to the manuscript, effectively restricted to rewording the title, and several minor text changes.			reprodmed3020015_perova	0
+The XGM2016 model is parameterized as a spherical harmonic series expansion resolved to degree and order (d/o) 719, which is the maximum resolution supported by the 15′ terrestrial gravity grid and a satellite-only model GOCO05s. For XGM2016, a significant focus is the optimal combination of the new terrestrial data with the latest satellite gravity information. The combination is based on the rigorous solution of a full normal equation system up to the maximum d/o 719. The calculation of the XGM2019 spheroidal harmonic model coefficients up to d/o 719 consists of a weighted least squares adjustment of GOCO06s with the primary 15′ NGA ground gravity dataset. The XGM2016 and XGM2019 models used the 15′ NGA ground gravity dataset, however, the refined GFMs in this study are obtained by combining the GRACE/GOCE-based GGMs and EGM2008 model, The gravity field information of 5′ terrestrial gravity data in EGM2008 is fully utilized. In addition, to consider the influence of higher frequency gravity field signals caused by topography, the RTM is utilized to further compensate for the omission errors in the refined GGMs. In this study, the combination of the satellite-only GGM with the EGM2008 is based on a pure complementation of the spherical harmonic coefficients at a specific degree, We added comparisons for XGM2016 or XGM2019 models in Table 4, we can find that the refined GFMs outperform XGM2016 and XGM2019 as well, the major improvement of the refined GFMs can be attributed to the GOCE data and topography signals. Please refer to page 14, Line 435-457.	2	1	Importantly, even though all local height datums are re- 42 lated to the MSL, the vertical offsets between them may be up to 2 m at global scale [1].			rs14061437_makarova	0
+"Thank you for your good question and advice. the high-quality GNSS/levelling-based height anomalies are used to check the refined GGMs for obtaining the optimal combination degrees. Thus, the refined GGMs provide better local quasi-geoid results. We have added descriptions and contents in the revised manuscript. The combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree in this paper, which is different from rigorous combination that is done on the basis of the normal equations and co-variance by a least-squares. The purpose of this pure combination can provide better local quasi-geoid results, it is said that it can obtain the characteristics of spatial ""localization"" for quasi-geoid. Combining the satellite-only GFM with EGM2008 by applying some sophisticated weighting approach usually combines the maximum degree and order of the satellite-only GFM and the EGM2008. Because the degree errors of the satellite-only GFM increase with the increase in degree and order, the noise starts to dominate the signals at high degree and order. The noise of satellite-only GFM maybe introduce by sophisticated weighting approach. In addition, the rigorous combination based on the sophisticated weighting approach usually needs the full error variance-covariance matrix of the spherical harmonic coefficients, but, the full error variance-covariance matrix of the spherical harmonic coefficients might generally not be available. Although the obtained results are already quite promising, it can be expected that the refined GFMs provide a guidance for determining the quasi-geoid or the geopotential value of the vertical datum in China. However, the combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree. However, such a procedure might cause a spectral gap between both models. In the next step, the rigorous combination or a smooth transition (such as: using hanning window) will be considered to derived the refined GGMs. Please refer to page 17, Line 544-545."	2	1	I have a few minor points (marked in the attached PDF) and two general points, which I like you to address.			rs14061437_makarova	0
+Please refer to page 2, Line 60.	2	1	Please comment on that question quantitatifely and qualitatively.			rs14061437_makarova	0
+Please refer to page 2, Line 63.	2	1	I have a few minor points (marked in the attached PDF) and two general points, which I like you to address.			rs14061437_makarova	0
+Please refer to page 11, Line 359.	2	1	Could you please comment on the effects you introduce by applying such a sharp truncation combination approach.			rs14061437_makarova	0
+Thank you for your advice. We have rewritten the mean values. Please refer to page 16, Line 496.	2	1	What is the main methodological difference and advantage of your method?			rs14061437_makarova	0
+Thank you for your comment. The unified topo data and Equation are used to compute RTM quasi-geoid height. We have added some expressions about the using of unified topo data and equations, which makes us easily misunderstand about the calculation process in the text. Please refer to page 12, Line 402.	2	1	Models Max Min Mean STD EIGEN-6C4 1.007 -1.696 0.048 0.187 GECO 1.579 -1.703 0.041 0.223 SGG-UGM-1 1.003 -1.671 0.052 0.194 SGG-UGM-2 1.003 -1.704 0.051 0.191 XGM2016 1.016 -1.757 -0.020 0.214 XGM2019 1.705 -1.737 0.081 0.213 Point 6: Your method for combining the satellite-only GFM with EGM2008 (sec.			rs14061437_makarova	0
+Thank you. Done. Please refer to page 8, Line 259.	2	1	If you have any information, please don’t hesitate to let us know.			rs14061437_makarova	0
+Thank you for your constructive suggestion. We have moved the discussion part in results to Discussion. The conclusions are recompiled and added according to your suggestions. Please refer to Line 498-550 and Line 551-589.	2	1	43 This is due to the fact that the MSL presents geographical and time-dependent variations, 44 Citation: Lastname, F.; Lastname, F.; Lastname, F. Title.			rs14061437_makarova	0
+Please refer to page 2, Line 60.	2	1	However, the 36 ellipsoidal height is not related to the Earth's gravity field.			rs14061437_perova	0
+Please refer to page 2, Line 63.	2	1	The goal of this paper is to derive 13 the geopotential value for the Chinese height datum (CNHD) in order to realize the height datum 14 unification in China.			rs14061437_perova	0
+Please refer to page 11, Line 359.	2	1	Thus, the refined GGMs provide better local quasi-geoid results.			rs14061437_perova	0
+Thank you for your advice. We have rewritten the mean values. Please refer to page 16, Line 496.	2	1	The refined GFMs are evaluated by using high-quality GNSS/lev- 22 elling data, the results show that the quasi-geoid accuracy of the refined DIR_R6_EGM2008_RTM 23 model in China has the optimal accuracy, and compared with the EGM2008 model and the DIR_R6 24 model, this refined model in China is improved by 9.6 cm and 21.8 cm, and the improvement ranges 25 are 35.7% and 55.8%, respectively.			rs14061437_perova	0
+The XGM2016 model is parameterized as a spherical harmonic series expansion resolved to degree and order (d/o) 719, which is the maximum resolution supported by the 15′ terrestrial gravity grid and a satellite-only model GOCO05s. For XGM2016, a significant focus is the optimal combination of the new terrestrial data with the latest satellite gravity information. The combination is based on the rigorous solution of a full normal equation system up to the maximum d/o 719. The calculation of the XGM2019 spheroidal harmonic model coefficients up to d/o 719 consists of a weighted least squares adjustment of GOCO06s with the primary 15′ NGA ground gravity dataset. The XGM2016 and XGM2019 models used the 15′ NGA ground gravity dataset, however, the refined GFMs in this study are obtained by combining the GRACE/GOCE-based GGMs and EGM2008 model, The gravity field information of 5′ terrestrial gravity data in EGM2008 is fully utilized. In addition, to consider the influence of higher frequency gravity field signals caused by topography, the RTM is utilized to further compensate for the omission errors in the refined GGMs. In this study, the combination of the satellite-only GGM with the EGM2008 is based on a pure complementation of the spherical harmonic coefficients at a specific degree, We added comparisons for XGM2016 or XGM2019 models in Table 4, we can find that the refined GFMs outperform XGM2016 and XGM2019 as well, the major improvement of the refined GFMs can be attributed to the GOCE data and topography signals. Please refer to page 14, Line 435-457. Table 4. Statistics of the height anomaly differences between GNSS/levelling and six higher-degree GFMs. Unit: (m).	2	1	The ellipsoid height relative to a given geo- 35 centric ellipsoid can be obtained quickly and accurately by using GNSS.			rs14061437_perova	0
+"Thank you for your good question and advice. the high-quality GNSS/levelling-based height anomalies are used to check the refined GGMs for obtaining the optimal combination degrees. Thus, the refined GGMs provide better local quasi-geoid results. We have added descriptions and contents in the revised manuscript. The combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree in this paper, which is different from rigorous combination that is done on the basis of the normal equations and co-variance by a least-squares. The purpose of this pure combination can provide better local quasi-geoid results, it is said that it can obtain the characteristics of spatial ""localization"" for quasi-geoid. Combining the satellite-only GFM with EGM2008 by applying some sophisticated weighting approach usually combines the maximum degree and order of the satellite-only GFM and the EGM2008. Because the degree errors of the satellite-only GFM increase with the increase in degree and order, the noise starts to dominate the signals at high degree and order. The noise of satellite-only GFM maybe introduce by sophisticated weighting approach. In addition, the rigorous combination based on the sophisticated weighting approach usually needs the full error variance-covariance matrix of the spherical harmonic coefficients, but, the full error variance-covariance matrix of the spherical harmonic coefficients might generally not be available. Although the obtained results are already quite promising, it can be expected that the refined GFMs provide a guidance for determining the quasi-geoid or the geopotential value of the vertical datum in China. However, the combination of the satellite-only GGM with the EGM2008 in this study is based on a pure complementation of the spherical harmonic coefficients at a specific degree. However, such a procedure might cause a spectral gap between both models. In the next step, the rigorous combination or a smooth transition (such as: using hanning window) will be considered to derived the refined GGMs. Please refer to page 17, Line 544-545."	2	1	We have added descriptions and contents in the revised manuscript.			rs14061437_perova	0
+Thank you for your comment. The unified topo data and Equation are used to compute RTM quasi-geoid height. We have added some expressions about the using of unified topo data and equations, which makes us easily misunderstand about the calculation process in the text. Please refer to page 12, Line 402.	2	1	It has improved the readability, clarity, and quality of our manuscript.			rs14061437_perova	0
+Thank you. Done. Please refer to page 8, Line 259.	2	1	Response to Reviewer 1 Comments: We thank you for your constructive and detailed comments.			rs14061437_perova	0
+Thank you for your constructive suggestion. We have moved the discussion part in results to Discussion. The conclusions are recompiled and added according to your suggestions. Please refer to Line 498-550 and Line 551-589.	2	1	Email: zcli@cumtb.edu.cn 9 4 National Geomatics Center of China, Beijing 100830, China; zhangpeng@ngcc.cn; ys@ngcc.cn 10 * Correspondence: zcli@cumtb.edu.cn;Tel.			rs14061437_perova	0
+I added a table listing the baseline characteristics, including BMI and smoking status, of subjects.	2	1	Comments 5 - I recommend deleting graphs 5, 8, and 11 because graphs 7, 10, and 13 show the same information together with lines obtained by analyzing the structural change of the time series.			s22072682_makarova	0
+Since this study focuses on AT search, detailed estimation of sample size is not performed. I added in data analysis section.	2	1	Comments 3 Please add ethics protocol approval number.			s22072682_makarova	0
+Project identification code is H29-4. I added in 2.1.subjects and 6.patents.	2	1	Comments 3 Please add ethics protocol approval number.			s22072682_makarova	0
+My manuscript is checked by an English proofreader. I will attach an English proofreading certificate.	2	1	"Dear Authors: I appreciate the possibility to review the paper entitled ""Exploration of an inflection point of ventilation parameters with anaerobic threshold using strucchange"" by the authors Aida et al."			s22072682_makarova	0
+I described the paper's aim.	2	1	1 Response to Reviewer 2 Comments Comments 1 -The paper's aim should be the same in the abstract and the introduction.			s22072682_makarova	0
+I redrew Figures 1 and 2 and described them as an adaptation of the original publication.	2	1	Comments 4 - In figure 3, the indication of the test time appears in the reverse direction.			s22072682_makarova	0
+I described baseline load of the participants in 2.2. Experimental Protocol and Setup.	2	1	"Dear Authors: I appreciate the possibility to review the paper entitled ""Exploration of an inflection point of ventilation parameters with anaerobic threshold using strucchange"" by the authors Aida et al."			s22072682_makarova	0
+I revised Figure 3.	2	1	Subject BMI Smoking history A 23.2 Non-smoker B 19.8 Non-smoker C 19.5 Non-smoker D 19.7 Non-smoker E 21.5 Non-smoker F 22.8 Non-smoker G 20 Non-smoker H 24.9 Non-smoker I 18.2 Non-smoker J 21.8 Non-smoker K 18.7 Non-smoker L 21.4 Non-smoker M 21.4 Non-smoker N 18.2 Non-smoker O 18.5 Non-smoker P 19.8 Non-smoker Q 20.5 Non-smoker mean 21 - SD 1.9 - Comments 2 Did authors estimate sample size?			s22072682_makarova	0
+I deleted graphs 5, 8, and 11.	2	1	Subject BMI Smoking history A 23.2 Non-smoker B 19.8 Non-smoker C 19.5 Non-smoker D 19.7 Non-smoker E 21.5 Non-smoker F 22.8 Non-smoker G 20 Non-smoker H 24.9 Non-smoker I 18.2 Non-smoker J 21.8 Non-smoker K 18.7 Non-smoker L 21.4 Non-smoker M 21.4 Non-smoker N 18.2 Non-smoker O 18.5 Non-smoker P 19.8 Non-smoker Q 20.5 Non-smoker mean 21 - SD 1.9 - Comments 2 Did authors estimate sample size?			s22072682_makarova	0
+We have improved the section to clarify the need to extend this line of research.	2	1	We have tried to address all of your comments and take action on as many as we could.			su14020860_makarova	0
+Thank you for your suggestion. From a theoretical point of view, we find various references that explain how CSR index is calculated as the ratio between the aggregation of 140 items focused on environmental, social and economic issues and the total number of items analyzed, which codes as 1 if the firm disclose the CSR information related each item, and 0. Gallego‐Álvarez, I., & Pucheta‐Martínez, M. C. Corporate social responsibility reporting and corporate governance mechanisms: An international outlook from emerging countries. Business Strategy & Development 2020, 3(1), 77–97. Pucheta-Martínez, M. C., & Gallego-Álvarez, I. (2021). The Role of CEO Power on CSR Reporting: The Moderating Effect of Linking CEO Compensation to Shareholder Return. Sustainability, 13(6), 3197. Moreover, we include a Table 1 to justify the items used in the Corporate social responsibility disclosure. Additionally, we have also introduced in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. We had the firm size variable labelled as LTA which is calculated as logarithm of total assets. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	Author Response Dear Reviewer 3, Thank you for your kind comments.			su14020860_makarova	0
+Thanks for your suggestion. I have improved the figure in line with academic norms.	2	1	Best regards, The authors Author Response File: Author Response.docx			su14020860_makarova	0
+Thank you very much for your words. In our modest opinion, the originality of the paper is the analysis of the moderating role played by the proportion of independent directors on boards of directors with the relationship between the constitution of remuneration committees and CSR disclosure. -We have not found previous research focused on this topic using independent directors as moderator variable. We have improved the introduction by adding a paragraph that better motivates the work. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclosure of CSR information due to the scarce existence of previous literature in recent years.	2	1	We are hopeful you find our responses/edits to your concerns (expressed below) sufficient.			su14020860_makarova	0
+Thank you very much for this suggestion since, indeed, the introduction was presented with scant motivation. We have corrected this issue and we have included a new paragraph based on remuneration committee and which explains the importance of including this committee for companies. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclsoure of CSR information due to the scarce existence of previous literature in recent years.	2	1	However, I accept this article only for the moderating variable the authors have tested.			su14020860_makarova	0
+In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the theoretical background. In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the section to clarify the need to extend this line of research and have included the following references: Abeysekera, I. (2012). Role of remuneration committee in narrative human capital disclosure. Accounting & Finance, 52, 1-23. Alotaibi, K. O., & Hussainey, K. (2016). Determinants of CSR disclosure quantity and quality: Evidence from non-financial listed firms in Saudi Arabia. International Journal of Disclosure and Governance, 13(4), 364-393. Chhaochharia, V. & Grinstein, Y. (2009). CEO compensation and board structure. The Journal of Finance, 61,(1), 231-261. Kanapathippillai, S., Mihret, D., & Johl, S. (2019). Remuneration committees and attribution disclosures on remuneration decisions: Australian evidence. Journal of business ethics, 158(4), 1063–1082 Suttipun, M. (2021). The influence of board composition on environmental, social and governance (ESG) disclosure of Thai listed companies.	2	1	Best regards, The authors Author Response File: Author Response.docx			su14020860_makarova	0
+Thank you for your suggestion, we have included in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	Dear Reviewer 1, We are sending you the second version of our paper.			su14020860_makarova	0
+Thank you for this suggestion. In our modest opinion, this paper presents the exhaustive revision and calculation.	2	1	Please, notice that our responses are provided in bold after your comments/suggestions.			su14020860_makarova	0
+Thank you. In the implications paragraph located in the conclusions section, we have added a short text to assess the practical implications.	2	1	International Journal of Disclosure and Governance, 1-12.			su14020860_makarova	0
+We have improved the section to clarify the need to extend this line of research.	2	1	Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			su14020860_perova	0
+Thank you for your suggestion. From a theoretical point of view, we find various references that explain how CSR index is calculated as the ratio between the aggregation of 140 items focused on environmental, social and economic issues and the total number of items analyzed, which codes as 1 if the firm disclose the CSR information related each item, and 0. Gallego‐Álvarez, I., & Pucheta‐Martínez, M. C. Corporate social responsibility reporting and corporate governance mechanisms: An international outlook from emerging countries. Business Strategy & Development 2020, 3(1), 77–97.	2	1	Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			su14020860_perova	0
+Thanks for your suggestion. I have improved the figure in line with academic norms.	2	1	We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			su14020860_perova	0
+Thank you very much for your words. In our modest opinion, the originality of the paper is the analysis of the moderating role played by the proportion of independent directors on boards of directors with the relationship between the constitution of remuneration committees and CSR disclosure. -We have not found previous research focused on this topic using independent directors as moderator variable. We have improved the introduction by adding a paragraph that better motivates the work. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclosure of CSR information due to the scarce existence of previous literature in recent years.	2	1	Please, notice that our responses are provided in bold after your comments/suggestions.			su14020860_perova	0
+Authors: Thank you very much for this suggestion since, indeed, the introduction was presented with scant motivation. We have corrected this issue and we have included a new paragraph based on remuneration committee and which explains the importance of including this committee for companies. Furthermore, we have included these other references to improve the introduction of the work: Code, U. G. G. (2006). Unified Good Governance Code of Listed Companies. Comisión Nacional del Mercado de Valores. Spain. Conyon, M. J., & Peck, S. I. (1998). Board control, remuneration committees, and top management compensation. Academy of management journal, 41(2), 146-157. Good Governance Code of Listed Companies (2020). Comisión Nacional del Mercado de Valores. Spain. Fama, E.F., & Jensen, M.C. (1983). Separation of ownership and control. Journal of Law and Economics, 24, 301–325. Harrison, J. R. (1987). The strategic use of corporate board committees. California Management Review, 30(1), 109-125. Khan, A., Muttakin, M. B., & Siddiqui, J. (2013). Corporate governance and corporate social responsibility disclosures: Evidence from an emerging economy. Journal of business ethics, 114(2), 207-223. Tao, N. B., & Hutchinson, M. (2013). Corporate governance and risk management: The role of risk management and compensation committees. Journal of Contemporary Accounting and Economics, 9(1), 83-99. Williamson, O. E. (1983). Credible commitments: Using hostages to support exchange. The American Economic Review, 73(4), 519–540. Zaid, M. A., Abuhijleh, S. T., & Pucheta‐Martínez, M. C. (2020). Ownership structure, stakeholder engagement, and corporate social responsibility policies: The moderating effect of board independence. Corporate Social Responsibility and Environmental Management, 27(3), 1344-1360. -We have justified the need to extend the literature focused on the compensation committees and the disclsoure of CSR information due to the scarce existence of previous literature in recent years.	2	1	Please, notice that our responses are provided in bold after your comments/suggestions.			su14020860_perova	0
+In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the theoretical background. In our modest opinion, the originality of the paper is: (i) analyse of the association between remuneration committees and CSR disclosure; (ii) examine the moderating effect of independent board in the association between the remuneration committees and the disclosure of CSR information. We have not found previous research focused on these both topics. We have improved the section to clarify the need to extend this line of research and have included the following references: Abeysekera, I. (2012). Role of remuneration committee in narrative human capital disclosure. Accounting & Finance, 52, 1-23. Alotaibi, K. O., & Hussainey, K. (2016). Determinants of CSR disclosure quantity and quality: Evidence from non-financial listed firms in Saudi Arabia. International Journal of Disclosure and Governance, 13(4), 364-393. Chhaochharia, V. & Grinstein, Y. (2009). CEO compensation and board structure. The Journal of Finance, 61,(1), 231-261. Kanapathippillai, S., Mihret, D., & Johl, S. (2019). Remuneration committees and attribution disclosures on remuneration decisions: Australian evidence. Journal of business ethics, 158(4), 1063–1082 Suttipun, M. (2021). The influence of board composition on environmental, social and governance (ESG) disclosure of Thai listed companies. International Journal of Disclosure and Governance, 1-12.	2	1	Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			su14020860_perova	0
+In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Authors: Thank you for your suggestion, we have included in the control variable paragraph more information to provide justifications on why those control variables are considered in this study. In this sense, we have used the control variables in line with previous research to avoid biased results. We have introduced the following references: Ali, W., Frynas, J., & Mahmood, Z. (2017). Determinants of corporate social responsibility (CSR) disclosure in developed and developing countries: A literature review. Corporate Social Responsibility and Environmental Management,24, 273–294 Barako, D., & Brown, A. (2008). Corporate social reporting and board rep-resentation: Evidence from the Kenyan banking sector. Journal of Management and Governance,12, 309–324 Brammer, S., Millington, A., & Pavelin, S. (2007). Gender and ethnic diversity among UK corporate boards. Corporate Governance: An International Review,15(2), 393–403 Chau, G., & Gray, S. (2010). Family ownership, board independence and voluntary disclosure: Evidence from Hong Kong. Journal of International Accounting, Auditing and Taxation,19(2), 93–109 Donnelly, R., & Mulcahy, M. (2008). Board structure, ownership, and voluntary disclosure in Ireland. Corporate Governance: An International Review,16(5), 416–429 Jizi, M., Salama, A., Dixon, R., & Stratling, R. (2014). Corporate governance and corporate social responsibility disclosure: Evidence from the US banking sector. Journal of Business Ethics,125(4), 601–615 Haniffa, R., & Cooke, T. (2005). The impact of culture and governance on corporate social reporting. Journal of Accounting and Public Policy,2 4(5), 391–430 Liao, L., Luo, L., & Tang, Q. (2015). Gender diversity, board independence, environmental committee, and greenhouse gas disclosure. The British Accounting Review, 47, 409–424 Reverte, C. (2009). Determinants of corporate social responsibility disclosure ratings by Spanish listed firms. Journal of Business Ethics,8 8,351–366	2	1	We had the firm size variable labelled as LTA which is calculated as logarithm of total assets.			su14020860_perova	0
+Thank you for this suggestion. In our modest opinion, this paper presents the exhaustive revision and calculation.	2	1	Best regards, The authors Author Response File: Author Response.docx			su14020860_perova	0
+Thank you. In the implications paragraph located in the conclusions section, we have added a short text to assess the practical implications.	2	1	The results may not be robust without these controls.			su14020860_perova	0
+Our paper has been substantially revised, and we have addressed the five suggested points for improvements by reviewer 1 (see below for more detail).	2	1	10 Reviewer 1: Fourth, the section on co-evolution warrants serious re-thinking.			su14052976_makarova	0
+Thanks to a substantial revision our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions.	2	1	This also takes away the need for an overly elaborate introductory lit review.			su14052976_makarova	0
+We have now provided a substantive elaboration on the Natural Social Contract (see section 3), and how contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). More specifically, we have provided more detail on transformative social-ecological innovation (see also the new figure 1 on TSEI), on the mutual gains approach and institutional design principles (see new table 1).	2	1	4 Reviewer 2: “2) In this sense, I recommend to start with the broader discussion of the natural contract (beggining of session 3), ending with the question of how to put in practice this social contract, what challenges (what is the main focus of this text).			su14052976_makarova	0
+The line-up of the figures (no.	2	1	One wonders whether the notion of transformative change really warrant this evolutionary perspective.			su14052976_makarova	0
+We moved up the discussion on governance and steering and elaborated on the generative mechanisms behind a transition to a Natural Social Contract, e.g. visualized by the TSEI framework below, with more attention on three different models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.	2	1	2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers.			su14052976_makarova	0
+The question of how the NSC can help to meet and overcome such resistances has been addressed by diving into governance approaches that are capable of dealing with complexity, ambiguity and uncertainty, and with particular attention for multiple value creation and the mutual gains approach (e.g. in section 2). At the same time, we are careful not to present those approaches as simple solutions to those problems.	2	1	"The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial."			su14052976_makarova	0
+has been revised	2	1	We have now provided a substantive elaboration on the Natural Social Contract (see section 3), and how contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent).			su14052976_makarova	0
+We have taken out the statement that a NCS is a master signifier and rechecked the English.	4	1	Indeed, it is partly a manifesto, but it also offers an action perspective in addition to an analytical perspective.			su14052976_makarova	0
+Below we will respond to the suggestions by reviewer 2	2	1	In our revised contribution we believe more attention is given to limits of steering and unanticipated results through references to the literature about this.			su14052976_makarova	0
+Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance. For details see section 2 on page 3-10.	2	1	Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence.			su14052976_makarova	0
+"cf. In the given circumstances, and for the given topic, this seems very appropriate. One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive. Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence. We agree that the ""normative-prescriptive"" tone and use of ""should be"" was excessive. This has been changed where appropriate. At the same time, it is unavoidable in a paper which is partly a manifesto., in the form of a call for the creation of a Natural Social Contract and for transformation-oriented forms of governance. We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use. The need for critical analysis and the presence of limitations to steering are acknowledged. editor’s comment 3 about being normative: First of all, we would like to underline that it is ok to be normative."	2	1	15 Reviewer 2: 5) Formal aspects: The text needs a language revision and formatting of the figures to make them more readable.			su14052976_makarova	0
+The systemic leverage points for transformation offered can be used for achieving change through questions about interdependencies which can be utilised by actor coalitions interested in transformative change through sustainability transitions which are not unduly sectoral but also pay attention to issues of just transition, resilience and the overarching goal of instituting a Natural Social Contract. Achieving this requires and involves a rebalancing of society and new imaginaries (as master signifiers), such as the well-being economy and a Natural Social Contract, as important new orientations.	2	1	"The article brings a current theoretical discussion, putting into dialogue the debates of sustainability and public governance, defending the idea of a co-evolutionary governance that can put into practice a ""Natural Contract"", as proposed by the authors."			su14052976_makarova	0
+We fully agree that there is not a single path or model for building public governance that favors sustainability. In this regard, we have stressed, on several occasions, that social contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.	2	1	2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers.			su14052976_makarova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders. For more information see section 2 on page 3-10. Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance (including Ansell and Gash, 2015), evolutionary steering and adaptive governance.	2	1	One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive.			su14052976_makarova	0
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4). 2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers. Our line of argumentation has been revised, and examples have been reduced to examples. Thanks to many changes our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions. In addition, the line-up of the figures (no.	2	1	Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence.			su14052976_makarova	0
+We have improved the focus and structure in ways described below.	4	1	I suggest to improve the interface between the propositions presented and what comes before in the text.			su14052976_makarova	0
+Governance approaches mentioned are now related to Transformative Social-Ecological Innovation (TSEI), which are innovations with explicit transformative goals and attention to mutual gains, and originating from political science and negotiation theory in particular. EGT gives attention to actor/institutions dialectics and power/knowledge dialectics, elements that have taken been into account in the TSEI-framework (figure 1) and related text.	4	1	Therefore, it seems essential to me to make it clear in each of the sections of the article that deal with these notions from where they start and how they relate to previous debates to support the authors' arguments.			su14052976_makarova	0
+Our build-up of argumentation has been improved.	4	1	Many thanks for these constructive comments, and we agree with the suggestions for improvements made by the editor and reviewers.			su14052976_makarova	0
+Changes have been made accordingly. See section with new title: “Transformation pleas and governance approaches for achieving this”. The section now better relates/differentiates evolutionary governance from the other governance approaches mentioned. EGT understands governance as radically evolutionary and is mindful of limitations of any form of governance.	4	1	"I am referring to universal and generalist statements with a ""should be"" tone that do not fit well with a scientific article, like the one in lines 51 to 54, among many others in the article."			su14052976_makarova	0
+Practical examples in this section are now more explicit on co-evolutionary steering and multiple value creation insights. Social contract theory helps to recognize duties and rights of citizens and consider issues of rebalancing society, and approaches and mechanisms for achieving this.	4	1	Figure 4 - Possible leverage points and key variables for a transition to nature-inclusive agriculture & agro-ecology in the Netherlands (at regional level) (Huntjens et al 2022) 6 Reviewer 1: This paper seeks to develop the notion of a Natural Social Contract (NSC) to enable transformative governance based on co-evolution.			su14052976_makarova	0
+This comment is not entirely clear to us. The four proposals for transition policy in section 4 are based on co-evolutionary governance and related discussions in previous sections.	4	1	At the same time, we are careful not to present those approaches as simple solutions to those problems.			su14052976_makarova	0
+Language and figures have been improved.	4	1	We want to sincerely thank you for your interest in the Sustannability and for sharing your work with us.			su14052976_makarova	0
+Format of figures and tables has been improved where appropriate.	2	1	"15 Reviewer 2: 2) Remove from the problematization the ""normative-prescriptive"" tone starting from questions and nuisances that justify the study and from gaps in the literature."			su14052976_makarova	0
+Thanks to a substantial revision our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions.	2	1	The transition path in question is always context-specific, path-dependent and goal-dependent.			su14052976_perova	0
+We have now provided a substantive elaboration on the Natural Social Contract (see section 3), and how contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). More specifically, we have provided more detail on transformative social-ecological innovation (see also the new figure 1 on TSEI), on the mutual gains approach and institutional design principles (see new table 1).	2	1	All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			su14052976_perova	0
+The line-up of the figures (no.	2	1	The paper could do more to translate the general aspects of transformative governance into more specific project aspects, as an evaluative framework for debating the cases.			su14052976_perova	0
+We moved up the discussion on governance and steering and elaborated on the generative mechanisms behind a transition to a Natural Social Contract, e.g. visualized by the TSEI framework below, with more attention on three different models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.	2	1	Figure 2: Possible systemic leverage points for a societal transformation towards a Natural Social Contract, through transformative governance based on co-evolution across several interrelated dimensions (this figure is a synthesis of table 3.4 in Huntjens, 2021) Figure 3: The Intervention Flower as a tool for identifying options for co-evolutionary steering through connecting actor-coalitions and interdependent systemic leverage points.			su14052976_perova	0
+The question of how the NSC can help to meet and overcome such resistances has been addressed by diving into governance approaches that are capable of dealing with complexity, ambiguity and uncertainty, and with particular attention for multiple value creation and the mutual gains approach (e.g. in section 2). At the same time, we are careful not to present those approaches as simple solutions to those problems.	2	1	How does a project mission contribute to a broader social contract?			su14052976_perova	0
+has been revised	2	1	This paper is openly normative in the call for the creation of a Natural Social Contract and for transformation-oriented forms of governance, but the governance approaches advocated have been tried and tested, yielding insights for future use.			su14052976_perova	0
+We have taken out the statement that a NCS is a master signifier and rechecked the English.	4	1	Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.			su14052976_perova	0
+Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance. For details see section 2 on page 3-10.	2	1	Our line of argumentation has been revised, and examples have been reduced to examples.			su14052976_perova	0
+"cf. In the given circumstances, and for the given topic, this seems very appropriate. One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive. Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence. We agree that the ""normative-prescriptive"" tone and use of ""should be"" was excessive. This has been changed where appropriate. At the same time, it is unavoidable in a paper which is partly a manifesto., in the form of a call for the creation of a Natural Social Contract and for transformation-oriented forms of governance. We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use. The need for critical analysis and the presence of limitations to steering are acknowledged. editor’s comment 3 about being normative: First of all, we would like to underline that it is ok to be normative."	2	1	Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.			su14052976_perova	0
+The systemic leverage points for transformation offered can be used for achieving change through questions about interdependencies which can be utilised by actor coalitions interested in transformative change through sustainability transitions which are not unduly sectoral but also pay attention to issues of just transition, resilience and the overarching goal of instituting a Natural Social Contract. Achieving this requires and involves a rebalancing of society and new imaginaries (as master signifiers), such as the well-being economy and a Natural Social Contract, as important new orientations.	2	1	EGT gives attention to actor/institutions dialectics and power/knowledge dialectics, elements that have taken been into account in the TSEI-framework (figure 1) and related text.			su14052976_perova	0
+We fully agree that there is not a single path or model for building public governance that favors sustainability. In this regard, we have stressed, on several occasions, that social contract formation takes place in polycentric ways at different levels (always context-specific and path-dependent). Challenges related to sustainability, social inequalities, or public governance, regardless of whether it takes place in the North or South, call for transformation-oriented forms of governance which are capable of dealing with complexity, ambiguity, and uncertainty.	2	1	The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			su14052976_perova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders. For more information see section 2 on page 3-10. Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance (including Ansell and Gash, 2015), evolutionary steering and adaptive governance.	2	1	We would, however, like to frame this positively: we believe that if the nature of the paper and the chain of argumentation are further clarified, it can be a major contribution not just to several academic discussions (I would mention here the just transition discussions, resilience, innovation, co-evolution, governance, etc) but also to societal debates, to, hopefully, public discourse, leading into policy discourse.			su14052976_perova	0
+The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4). 2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers. Our line of argumentation has been revised, and examples have been reduced to examples. Thanks to many changes our paper now provides a clear argumentation and logical structure, with the first part delving into literature to explain the key issues and concepts, and then moving to solutions. In addition, the line-up of the figures (no.	2	1	I thank the opportunity to evaluate this article, which I really enjoyed reading.			su14052976_perova	0
+Format of figures and tables has been improved where appropriate.	2	1	Co-evolution is now associated with all kinds of change-in-tandem, social-sustainable, different institutional actors, discursive items, etc.			su14052976_perova	0
+We have improved the focus and structure in ways described below.	4	1	The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			su14052976_perova	0
+Governance approaches mentioned are now related to Transformative Social-Ecological Innovation (TSEI), which are innovations with explicit transformative goals and attention to mutual gains, and originating from political science and negotiation theory in particular. EGT gives attention to actor/institutions dialectics and power/knowledge dialectics, elements that have taken been into account in the TSEI-framework (figure 1) and related text.	4	1	What remains unclear and implicit, however, is the form and operationalisation of such contract.			su14052976_perova	0
+Our build-up of argumentation has been improved.	4	1	12 Reviewer 1: Source citations are sometimes unclear, as they refer to “cf.” items and lack page numbers Has been revised 13 Reviewer 2: I thank the opportunity to evaluate this article, which I really enjoyed reading.			su14052976_perova	0
+Changes have been made accordingly. See section with new title: “Transformation pleas and governance approaches for achieving this”. The section now better relates/differentiates evolutionary governance from the other governance approaches mentioned. EGT understands governance as radically evolutionary and is mindful of limitations of any form of governance.	4	1	Much of this, moreover, is action-oriented, pointing out major carriers and direction of change.			su14052976_perova	0
+Practical examples in this section are now more explicit on co-evolutionary steering and multiple value creation insights. Social contract theory helps to recognize duties and rights of citizens and consider issues of rebalancing society, and approaches and mechanisms for achieving this.	4	1	Two, the nature and form of evolutionary variation, selection and dynamics remain unclear.			su14052976_perova	0
+This comment is not entirely clear to us. The four proposals for transition policy in section 4 are based on co-evolutionary governance and related discussions in previous sections.	4	1	This would make room to bring session 02 that addresses evolutionary/transformative governance to the TSEI.” Our build-up of argumentation has been improved.			su14052976_perova	0
+Language and figures have been improved.	4	1	The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary.			su14052976_perova	0
+The confusingly written part has been simplified again. In addition, in order to solve the problem of awkward English, we have re-translated the entire sentence.	2	1	Therefore, it seems that the contribution breaks down to proposing strict tracking services which feasibility and acceptance by the population is not assessed.			su14052981_makarova	0
+As your comments, Korea's smart city has technological and normative advances compared to other countries. However, we think that the USA and China are more developed in service than Korea`s (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022). Therefore, this paper intends to cover only general smart city technologies worldwide (Smart city trends: A focus on 5 countries and 15 companies, Cities 2022). We fully agree with the reviewer's comments that they may feel uncomfortable in other countries. After that, we will do more research and expand the case to other countries. Thank you very much for your comments.	2	1	If it goes beyond COVID, this is a claim that needs to be backed with a sound and transparent evaluation by the authors or literature.			su14052981_makarova	0
+4. In this paper, the monitoring of sewage and wastewater for population level assessment of infection mentioned in smart city applications and urban automation was added as a limitation of this study. In the opinion of reviewers, epidemics in cities are not simply spread by one factor. Therefore, various factors should be dealt with, but in this paper, only heat and movement lines, which were mainly dealt with in Korea. The added part is as follows. Conclusion: “… Also, epidemics in cities are not spread simply by one factor. It is important to operate the monitoring system for sewage and wastewater for population level assessment of infection, which is considered to be the most important source of infection. 2 Therefore, they should be considered in future research.	2	1	This is a very important point, since it changes what is the contribution of the paper.			su14052981_makarova	0
+Additionally, smart city services could respond neither timely nor sequentially.	2	1	Table 4 presents smart city projects of several different countries.			su14052981_makarova	0
+This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services.	2	1	I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable.			su14052981_makarova	0
+Second, recommendations on combining or dismissing certain smart city services, as per the needs of coping with COVID-19, are summarized. Third, smart city services must be utilized only for addressing pandemics, as data from the healthcare system consists of personal information. Therefore, smart city services for responding to COVID-19 must be flexible.	2	1	All these issues could have been picked up by a simple check.			su14052981_makarova	0
+The source has been specified. - (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022).	2	1	Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2.			su14052981_makarova	0
+We have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression 'EA' is a unit used only in certain countries.	2	1	I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations.			su14052981_makarova	0
+Thanks for your kind comments.	2	1	It the threats concern COVID only, it does not really make sense, as many of the mentioned services simply do not have this goal (e.g.			su14052981_makarova	0
+We also agree with reviewers that it's an ideal situation to use built-in services only when needed and turn them off when not needed. Therefore, in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport(Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data.	2	1	In addressing the comments, I have been able to improve the clarity and content of the manuscript vastly.			su14052981_makarova	0
+We rechecked the paper related to “Key missing components/literatures”. We sincerely apologize for repeating basic mistakes. They all have been revised and reflected.	2	1	(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen?			su14052981_makarova	0
+Thanks for your kind words. The quality of the paper has improved on the reviewer's advice. There are many things that need to be improved in this paper, however we will remember your sincere advices and write more advanced papers later. Thanks again.	4	1	Indeed, the phases in a Texan county are of no use to understand the authors’ contribution.			su14052981_makarova	0
+- This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.	4	1	They could easily break the section down to the essential information that would be the WHO phases and the Korean healthcare phases, and mention that there are global and local plans throughout the world with different phases.			su14052981_makarova	0
+In order to improve the quality of the thesis, professional translation has been carried out again. We sincerely apologize for the rudimentary mistakes related to some expressions and errors.	2	1	I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable.			su14052981_makarova	0
+References are rechecked and deleted or erroneous references are recited or deleted. We humbly accept the reviewer's opinion that there are many references, and we have deleted references that are deemed unnecessary. We humbly accept the reviewer's comments that there is a quality problem, and We did our best to double-check, revise, and improve the quality of our papers.	2	1	(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen?			su14052981_makarova	0
+We fully agree with the reviewer's comment that integration between services is difficult not only in smart cities but also in other fields. As advised by the reviewer, the situation in Korea was described in Figure 5 (Outbreak of COVID-19 in Korea and the limitations of smart city services) and Table 10 (Limitations of smart city services in each phase). The spread of COVID cannot simply be blocked by smart city services, but by simplifying the current state and proceeding with the thesis, we did not reflect Korea's medical stage with geographic characteristics. We will conduct additional research in the future to fully reflect the reviewer's comments. Thanks for the advice.	2	1	I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations.			su14052981_makarova	0
+The 'EA' refers to the quantity, but all have been deleted. We sincerely apologize for the basic mistake. We will check in more detail later. We reviewed the literature review and deleted redundant or unnecessary content. Also, we have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression ‘EA’ is a unit used only in certain countries.	2	1	Several different plans are described, which is useless information for the reader.			su14052981_makarova	0
+Smart city projects are underway all over the world. Also, although smart city services differ by country, the smart city structure has a similar framework to a certain level. Therefore, if we can transform the structure of the smart city, we decided that it would be a great tool to cope with situations such as COVID, even a little.	2	1	[15], [22], [25]) it is impossible to retrieve the source.			su14052981_makarova	0
+Like the reviewer's comments, the rest of the table's contents other than those related to pandemic have been deleted.	2	1	line 213: “As mentioned in 0”) Finally, and most importantly, the format of the bibliography is highly problematic.			su14052981_makarova	0
+Sections 2.2 and 2.3 have identified errors with the same title. Accordingly, the title has been modified as follows. We apologize once again for repeating basic mistakes. - 2.2 Status of smart city services - 2.3 Status of Pandemic #9.	2	1	Please check especially the conclusion, this is hardly readable.			su14052981_makarova	0
+We completely agree with the reviewer's sharp points. Accordingly, the contents including Tables 8 and 9 were deleted and reorganized into World Health Organization (WHO), Control and Prevention (CDC) and Korea Centers for Disease Control (KCDC).	2	1	“Revised model of smart city service structure”.			su14052981_makarova	0
+Thanks for the reviewer's comments. The title of this section 3 has been changed according to the comments of the reviewers.	2	1	"There are still some ""Reference not found"" errors and line breaks when referring to figures in the text, but almost everything regarding the format has been fixed, which gives the revised paper a much more professional look than its previous version."			su14052981_makarova	0
+In the case of Korea, rather than finding the infected person in advance, the focus was on identifying and blocking the spread of the infected person. Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately. As a result, the critical point of being identified before an infectious disease spreads was missed. However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard. We agree with the reviewers' opinion that the biggest cause of this COVID-19 transmission is the rapid spread of infected people around them by moving to public places before they show symptoms. Reflecting the reviewer's opinion that it is not clear, the entire sentence has been revised and translated clearly.	2	1	(pg14) “such services should be flexibly used only during a national crisis and removed when the service is no longer needed.” Is this likely to happen?			su14052981_makarova	0
+First, an epidemiological surveyor should decide whether additional collection of personal information is needed. If the answer is yes, the official should seek approval from relevant authorities to get access to the data. For example, as for the location information, separate permission from the National Police Agency is required.” Source: http://www.molit.go.kr/english/USR/BORD0201/m_28286/DTL.jsp?id=eng_mltm_new&mode=view&idx=2931 3) Key missing components/literatures: We fully agree with the reviewers' comments. Therefore, as the limit of this paper, the limit of the ethical aspect was first specified, and in the situation where the service is not needed, the service is deleted in consideration of the ethical aspect in the structure of the smart city. Also in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport (Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data.	2	1	line 85) The reference confirmed that the error occurred when the formatting was moved.			su14052981_makarova	0
+Also, we really appreciate the accurate comments from the reviewers.	2	1	[15], [22], [25]) it is impossible to retrieve the source.			su14052981_makarova	0
+Thank you for the reviewer's heartfelt comments. The corrections to the reviewer's concerns are as follows: #2.	4	1	I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors.			su14052981_makarova	0
+We checked again that errors occurred. After fixing the errors, we rechecked it on others PC. We sincerely apologize for the continued errors. References deleted in the first review were deleted because they were deemed unnecessary. Thanks for the comments #3.	4	1	Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			su14052981_makarova	0
+We agree with the reviewer's comments. As advised by the reviewer, the content of the scenario was additionally reflected in the abstract. The contents are as follows.	4	1	It is also hard to read, as it contains more tabular content than text.			su14052981_makarova	0
+The description of the characteristics of each city shown in Table 3 was insufficient, as was the reviewer's opinion. Accordingly, the project described in Table 3 was briefly explained and the contents of the article were also revised. - “... As a representative smart city project in Korea, Pilot city, Regulatory sandbox and Smart City Challenge are in progress. To summarize the smart city services used in this project, medical, crime prevention, transportation and environmental services are the most used.” Table 7 also agrees with the reviewer's advice that it does not fit the current content. Accordingly, the contents of Table 7 were deleted and only the contents described in the text were left. Thanks for your comments.	4	1	Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.			su14052981_makarova	0
+Thank you. As commented by reviewer, the content of Section 3.1 includes the content of Section 2. However, for the 3.2 Expansion of smart city structure that we want to present, I think it is necessary to organize the contents of Section 2 through Section 3.1 and to draw limitations. Of course, it is also correct to understand the structure as the opinions of reviewer. We put a lot of thought into reflecting the opinions. We are sorry; however, we have decided that it is difficult to change the structure at the moment. We ask for your generous understanding of the current structure.	4	1	This may be because of the error message 3 in sourcing in the document I received.			su14052981_makarova	0
+Thank you for understanding a little more about our paper. Also, I totally agree with the reviewer's comments about the skepticism of readers. Accordingly, the legal considerations specified in the first review have been added to the text. Its contents are as follows. “… In Korea, disease-related smart city services are legally sanctioned to be used only when necessary to protect personal information.”	4	1	Science of the Total Environment, 736, 139631.			su14052981_makarova	0
+We are sorry for not being clear on the answer to the last review. This paper tried to present a concept for the smart city structure to cope with major situations such as COVID-19. Therefore, it was not possible to present a specific method of adding or removing a smart city system. Currently, smart city services in Korea are trying to implement a structure that adds and deletes when necessary, as in this paper (Reference #73). However, we apologize for not being able to give you a clear answer on this question. After observing the process of implementing the system, we will present a specific method in the thesis we plan to proceed later.	4	1	I would thus include it in Section 2 instead.			su14052981_makarova	0
+Many revisions have been made to this paper according to the reviewer's comments. Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers. In addition, we will use a more advanced paper by reflecting it in the ongoing research. Also, we sincerely appreciate the accurate comments from the reviewers.	4	1	The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV.			su14052981_makarova	0
+The reference confirmed that the error occurred when the formatting was moved. In addition, errors in the sentences have been reviewed and corrected. We apologize again for the rudimentary mistake.	2	1	I thus find it now less essential to discuss the practical aspects of service adding and removing.			su14052981_makarova	0
+In other words, the structure lacks flexibility	2	1	For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source.			su14052981_makarova	0
+The full terms of the abbreviation are reflected.	2	1	I think many cities and countries will not be comfortable with this approach.			su14052981_makarova	0
+The incorrect part of the year has been corrected. Thanks for your pointing out #6.	2	1	I think that is both a big oversight, and an opportunity for critics.			su14052981_makarova	0
+we sincerely apologize. There were many rudimentary mistakes. We corrected the errors by rechecking the paper.	2	1	(if so, needs sourcing), or a synthesis by the authors?			su14052981_makarova	0
+Missing or erroneous references were rechecked and corrected.	2	1	Please always explain abbreviations before use - e.g.			su14052981_makarova	0
+The confusingly written part has been simplified again. In addition, in order to solve the problem of awkward English, we have re-translated the entire sentence.	2	1	Author Response Please see the attachment Author Response File: Author Response.pdf			su14052981_perova	0
+As your comments, Korea's smart city has technological and normative advances compared to other countries. However, we think that the USA and China are more developed in service than Korea`s (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022). Therefore, this paper intends to cover only general smart city technologies worldwide (Smart city trends: A focus on 5 countries and 15 companies, Cities 2022). We fully agree with the reviewer's comments that they may feel uncomfortable in other countries. After that, we will do more research and expand the case to other countries. Thank you very much for your comments.	2	1	It is well-known that different services are hard to integrate together (this is not specific to the smart city), but I would have liked the authors to illustrate in light of the pandemic context, and maybe to the context of Korea since their contribution is specific to this geographic context in that it integrates the 12 smart city categories of Korea and the healthcare phases of Korea.			su14052981_perova	0
+In this paper, the monitoring of sewage and wastewater for population level assessment of infection mentioned in smart city applications and urban automation was added as a limitation of this study. In the opinion of reviewers, epidemics in cities are not simply spread by one factor. Therefore, various factors should be dealt with, but in this paper, only heat and movement lines, which were mainly dealt with in Korea. The added part is as follows.	2	1	I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors.			su14052981_perova	0
+Additionally, smart city services could respond neither timely nor sequentially.	2	1	Table 7 does not bring much useful insights as well in its current form.			su14052981_perova	0
+This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services.	2	1	Please always explain abbreviations before use - e.g.			su14052981_perova	0
+Second, recommendations on combining or dismissing certain smart city services, as per the needs of coping with COVID-19, are summarized. Third, smart city services must be utilized only for addressing pandemics, as data from the healthcare system consists of personal information. Therefore, smart city services for responding to COVID-19 must be flexible.	2	1	Author Response Please see the attachment Author Response File: Author Response.pdf			su14052981_perova	0
+The source has been specified. - (Smart city trends: A focus on 5 countries and 15 companies, Cities, 2022).	2	1	We sincerely apologize for the basic mistake.			su14052981_perova	0
+We have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression 'EA' is a unit used only in certain countries.	2	1	Are there concrete examples of such systems being built and operated, then just turned off?			su14052981_perova	0
+Thanks for your kind comments.	2	1	I think that is both a big oversight, and an opportunity for critics.			su14052981_perova	0
+We also agree with reviewers that it's an ideal situation to use built-in services only when needed and turn them off when not needed. Therefore, in Korea, it is legally required to be used only when necessary. Ministry of Land, Infrastructure and Transport(Korea): “…… But the scope of data collected will be kept to minimum and a due procedure should be followed in acquiring the data. First, an epidemiological surveyor should decide whether additional collection of personal information is needed. If the answer is yes, the official should seek approval from relevant authorities to get access to the data. For example, as for the location information, separate permission from the National Police Agency is required.” Source: http://www.molit.go.kr/english/USR/BORD0201/m_28286/DTL.jsp?id=eng_mltm_new&mode=view&idx=2931 3) Key missing components/literatures: We rechecked the paper related to “Key missing components/literatures”.	2	1	Table 4 presents smart city projects of several different countries.			su14052981_perova	0
+Webb, W., & Toh, C. K. (2020). The smart city and COVID‐19. IET Smart cities, 2(2), 56-57. Inn, T. L. (2020). Smart city technologies take on COVID-19. World Health, 841. Sharifi, A., Khavarian-Garmsir, A. R., & Kummitha, R. K. R. (2021). Contributions of smart city solutions and technologies to resilience against the COVID-19 pandemic: a literature review. Sustainability, 13(14), 8018. Gusikhin, O. (2020). The Impact of COVID-19 Experience on Smart City and Future Mobility. In Smart Cities, Green Technologies, and Intelligent Transport Systems (pp. 308-321). Springer, Cham. Kim, H. M. (2021). Smart cities beyond COVID-19. In Smart Cities for Technological and Social Innovation (pp. 299-308). Academic Press. 4 Inn, T. L. (2020).	2	1	First, I have a series of concerns regarding the form.			su14052981_perova	0
+Farkas, K., Hillary, L. S., Malham, S. K., McDonald, J. E., & Jones, D. L. (2020). Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19. Current Opinion in Environmental Science & Health, 17, 14-20. Daughton, C. G. (2020). Wastewater surveillance for population-wide Covid-19: the present and future. Science of the Total Environment, 736, 139631. Bogler, A., Packman, A., Furman, A., Gross, A., Kushmaro, A., Ronen, A., ... & Bar-Zeev, E. (2020). Rethinking wastewater risks and monitoring in light of the COVID-19 pandemic. Nature Sustainability, 3(12), 981-990. Jaiswal, R., Agarwal, A., & Negi, R. (2020). Smart solution for reducing the COVID‐19 risk using smart city technology. IET Smart Cities, 2(2), 82-88.	2	1	Infected people are in the public space without knowing that they have COVID, since contagion happens before symptoms appears.			su14052981_perova	0
+Thanks for your kind words. The quality of the paper has improved on the reviewer's advice. There are many things that need to be improved in this paper, however we will remember your sincere advices and write more advanced papers later. Thanks again.	4	1	1 Dear Reviewer #1, I am grateful for your suggestions regarding my paper, and my answers to your comments are provided below.			su14052981_perova	0
+- This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.	4	1	I think it synthesizes some important and useful literatures, and has some useful points to make.			su14052981_perova	0
+In order to improve the quality of the thesis, professional translation has been carried out again. We sincerely apologize for the rudimentary mistakes related to some expressions and errors.	2	1	I would thus include it in Section 2 instead.			su14052981_perova	0
+References are rechecked and deleted or erroneous references are recited or deleted. We humbly accept the reviewer's opinion that there are many references, and we have deleted references that are deemed unnecessary. We humbly accept the reviewer's comments that there is a quality problem, and We did our best to double-check, revise, and improve the quality of our papers.	2	1	In addition, errors in the sentences have been reviewed and corrected.			su14052981_perova	0
+We fully agree with the reviewer's comment that integration between services is difficult not only in smart cities but also in other fields. As advised by the reviewer, the situation in Korea was described in Figure 5 (Outbreak of COVID-19 in Korea and the limitations of smart city services) and Table 10 (Limitations of smart city services in each phase). The spread of COVID cannot simply be blocked by smart city services, but by simplifying the current state and proceeding with the thesis, we did not reflect Korea's medical stage with geographic characteristics. We will conduct additional research in the future to fully reflect the reviewer's comments. Thanks for the advice.	2	1	The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV.			su14052981_perova	0
+The 'EA' refers to the quantity, but all have been deleted. We sincerely apologize for the basic mistake. We will check in more detail later. We reviewed the literature review and deleted redundant or unnecessary content. Also, we have checked and corrected the Author's initials that you pointed out. We also Cognized that the expression ‘EA’ is a unit used only in certain countries.	2	1	Author Response Please see the attachment Author Response File: Author Response.pdf			su14052981_perova	0
+Smart city projects are underway all over the world. Also, although smart city services differ by country, the smart city structure has a similar framework to a certain level. Therefore, if we can transform the structure of the smart city, we decided that it would be a great tool to cope with situations such as COVID, even a little.	2	1	Table 7 does not bring much useful insights as well in its current form.			su14052981_perova	0
+Like the reviewer's comments, the rest of the table's contents other than those related to pandemic have been deleted.	2	1	However, it misses a critical point of COVID.			su14052981_perova	0
+Sections 2.2 and 2.3 have identified errors with the same title. Accordingly, the title has been modified as follows. We apologize once again for repeating basic mistakes. - 2.2 Status of smart city services - 2.3 Status of Pandemic #9.	2	1	I think it synthesizes some important and useful literatures, and has some useful points to make.			su14052981_perova	0
+We completely agree with the reviewer's sharp points. Accordingly, the contents including Tables 8 and 9 were deleted and reorganized into World Health Organization (WHO), Control and Prevention (CDC) and Korea Centers for Disease Control (KCDC).	2	1	However, it misses a critical point of COVID.			su14052981_perova	0
+Thanks for the reviewer's comments. The title of this section 3 has been changed according to the comments of the reviewers.	2	1	(pg3) Tables 1 is interesting, but it’s not clear the source of the data.			su14052981_perova	0
+In the case of Korea, rather than finding the infected person in advance, the focus was on identifying and blocking the spread of the infected person. Therefore, Smart City Service checked the movement of the infected people by time, found close contacts in the vicinity, and took measures to self-quarantine immediately. As a result, the critical point of being identified before an infectious disease spreads was missed. However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard. We agree with the reviewers' opinion that the biggest cause of this COVID-19 transmission is the rapid spread of infected people around them by moving to public places before they show symptoms. Reflecting the reviewer's opinion that it is not clear, the entire sentence has been revised and translated clearly.	2	1	Smart solution for reducing the COVID‐19 risk using smart city technology.			su14052981_perova	0
+We fully agree with the reviewers' comments. Therefore, as the limit of this paper, the limit of the ethical aspect was first specified, and in the situation where the service is not needed, the service is deleted in consideration of the ethical aspect in the structure of the smart city. Also in Korea, it is legally required to be used only when necessary.	2	1	I have some theoretical questions about the applicability of these approaches outside of Korea.			su14052981_perova	0
+Also, we really appreciate the accurate comments from the reviewers.	2	1	Thank you again for your supportive review to improve the manuscript.			su14052981_perova	0
+Thank you for the reviewer's heartfelt comments. The corrections to the reviewer's concerns are as follows: #2.	4	1	(if so, needs sourcing), or a synthesis by the authors?			su14052981_perova	0
+We checked again that errors occurred. After fixing the errors, we rechecked it on others PC. We sincerely apologize for the continued errors. References deleted in the first review were deleted because they were deemed unnecessary. Thanks for the comments #3.	4	1	However, Korea has prevented a rapid collapse of the medical system by slowing the rate of propagation around it, and the role of smart city services has been significant in this regard.			su14052981_perova	0
+We agree with the reviewer's comments. As advised by the reviewer, the content of the scenario was additionally reflected in the abstract. The contents are as follows.	4	1	I have some theoretical questions about the applicability of these approaches outside of Korea.			su14052981_perova	0
+The description of the characteristics of each city shown in Table 3 was insufficient, as was the reviewer's opinion. Accordingly, the project described in Table 3 was briefly explained and the contents of the article were also revised. - “... As a representative smart city project in Korea, Pilot city, Regulatory sandbox and Smart City Challenge are in progress. To summarize the smart city services used in this project, medical, crime prevention, transportation and environmental services are the most used.” Table 7 also agrees with the reviewer's advice that it does not fit the current content. Accordingly, the contents of Table 7 were deleted and only the contents described in the text were left. Thanks for your comments.	4	1	I think many cities and countries will not be comfortable with this approach.			su14052981_perova	0
+Thank you. As commented by reviewer, the content of Section 3.1 includes the content of Section 2. However, for the 3.2 Expansion of smart city structure that we want to present, I think it is necessary to organize the contents of Section 2 through Section 3.1 and to draw limitations. Of course, it is also correct to understand the structure as the opinions of reviewer. We put a lot of thought into reflecting the opinions. We are sorry; however, we have decided that it is difficult to change the structure at the moment. We ask for your generous understanding of the current structure.	4	1	First, an epidemiological surveyor should decide whether additional collection of personal information is needed.			su14052981_perova	0
+Thank you for understanding a little more about our paper. Also, I totally agree with the reviewer's comments about the skepticism of readers. Accordingly, the legal considerations specified in the first review have been added to the text. Its contents are as follows.	4	1	Please check especially the conclusion, this is hardly readable.			su14052981_perova	0
+We are sorry for not being clear on the answer to the last review. This paper tried to present a concept for the smart city structure to cope with major situations such as COVID-19. Therefore, it was not possible to present a specific method of adding or removing a smart city system. Currently, smart city services in Korea are trying to implement a structure that adds and deletes when necessary, as in this paper (Reference #73). However, we apologize for not being able to give you a clear answer on this question. After observing the process of implementing the system, we will present a specific method in the thesis we plan to proceed later.	4	1	The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.			su14052981_perova	0
+Many revisions have been made to this paper according to the reviewer's comments. Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers. In addition, we will use a more advanced paper by reflecting it in the ongoing research. Also, we sincerely appreciate the accurate comments from the reviewers.	4	1	The authors than write that these services have a limitation in that cannot predict and prevent transmission of threats such as COVID.			su14052981_perova	0
+The reference confirmed that the error occurred when the formatting was moved. In addition, errors in the sentences have been reviewed and corrected. We apologize again for the rudimentary mistake.	2	1	Please always explain abbreviations before use - e.g.			su14052981_perova	0
+In other words, the structure lacks flexibility	2	1	Many revisions have been made to this paper according to the reviewer's comments.			su14052981_perova	0
+The full terms of the abbreviation are reflected.	2	1	This is a very important point, since it changes what is the contribution of the paper.			su14052981_perova	0
+The incorrect part of the year has been corrected. Thanks for your pointing out #6.	2	1	I will give a clearer example from p.15, line 345: Second, the ( Figure 4.			su14052981_perova	0
+we sincerely apologize. There were many rudimentary mistakes. We corrected the errors by rechecking the paper.	2	1	Thank you again for your supportive review to improve the manuscript.			su14052981_perova	0
+Missing or erroneous references were rechecked and corrected.	2	1	This may be because of the error message in sourcing in the document I received.			su14052981_perova	0
+We very much appreciate the contributions.	2	1	The authors test a popular assessment tool using confirmatory factor analysis and finds that the model is weak.			su14095590_perova	0
+An analysis of extreme values was carried out, using univariate and multivariate detection methods. The analysis resulted in the elimination of 34 univariate extreme values and 2 multivariate extreme values. All statistical analysis was repeated considering a database of 520 companies.	2	1	Very well structured, with a critical and current problem.			su14095590_perova	0
+Thank you for your suggestion. We researched more literature on the subject and adopted a more moderate perspective for assessing normality (|Sk|>2-3, |Ku|>7-10 and |KuMult|>10). As you mention, with the elimination of the extreme values, the skewness and kurtosis values improved significantly, approaching the range -1 to 1.	2	1	Table 2 and the paragraph after it is a repetition of Figure 1 and Table 1, could be cut.			su14095590_perova	0
+The discussion section has been improved and future research recommendations have been added in the conclusions.	2	1	We have made all the minor corrections indicated.			su14095590_perova	0
+The hypothesis was removed since, as mentioned, it doesn't state what we are trying to test in this study.	2	1	1 Response to Reviewer 4 Comments Point 1: The paper is well structured and presented in a very meaningful manner.			su14095590_perova	0
+Major changes have been made to the paper. We hope that this new version may deserve your recommendation for publication.	2	1	Even though the areas are from B Lab, the authors should study and related them to sustainability and value creation.			su14095590_perova	0
+The manuscript has been revised to improve grammar and english language.	4	1	2nd round Title: OK Abstract: OK Introduction: There is an improvement to do in the objective because it is still different from the one presented in the Abstract, and needs a correction to be aligned with the content of the text.			su14095590_perova	0
+The discussion and conclusions sections have been improved (text in blue).	4	1	In line 384 the authors should substitute ‘chapter’ for ‘section’.			su14095590_perova	0
+Thanks for the comment. The conclusions section has been improved (text in blue).	4	1	There isn’t a source below the Title of Figure 1, though after that seems that B Corporation is the owner of it.			su14095590_perova	0
+Thank you. We very much appreciate the comment.	2	1	Discussion: The authors made an effort to add more discussion, though no one was about Cronbach’s alpha.			su14095590_perova	0
+The argument associated with studies [28] & [29] was added.	2	1	No any future research recommendations have been suggested by the respected authors.			su14095590_perova	0
+The words Table and Figure have all been revised to start with a capital letter.	2	1	Point 2: Abstract: The content is well organized, but the results need some to attract the reader.			su14095590_perova	0
+Paragraph presented after Table 2 (page 7) has been moved to footnote.	2	1	The hypothesis is interesting when they come from a discussion in the literature review to the field research to test a theory (models, methods, frameworks, etc).			su14095590_perova	0
+• The (***) have been replaced by the p-values (<0.001) in Table - B Impact Assessment - Regression Weights (original model) (current Table 6).	2	1	It’s their signature in the text, so I understand that they deserve this recognition.			su14095590_perova	0
+Future research recommendations have been added to the conclusions.	2	1	The analysis is interesting, but demands more comparisons and explanations for cuts and adds.			su14095590_perova	0
+The manuscript has been revised. The literature review has been improved, and new references have been added (text in blue).	4	1	We believe that the reviewers' suggestions have been very helpful in improving the manuscript.			su14095590_perova	0
+The method section has been improved (text in blue).	4	1	The literature review has been improved, and new references have been added (text in blue).			su14095590_perova	0
+We believe that the reviewers' suggestions have been very helpful in improving the manuscript. Thank you for the comment. The manuscript has been entirely revised.	4	1	I strongly recommend the authors find the new one.			su14095590_perova	0
+Thank you for the suggestion. The title has been clarified in line with the main objective.	2	1	The authors should reflect the presence of that in the text.			su14095590_perova	0
+The results were improved and the paper's contribution was added.	2	1	Abstract: The main objective is to analyze the B Impact Assessment, verifying the robustness and consistency of the model to measure and improve the economic, social, and environmental impact of companies.			su14095590_perova	0
+The introduction has been improved, including a better explaination of B Impact Assessment. The use of confirmatory factor analysis has been explained. The aim of the paper was revised in the abstract to be aligned with the one presented in the introduction.	2	1	The authors should explain better what is B Impact Assessment.			su14095590_perova	0
+Lines 70 to 82 have been revised and included in the introduction. Lines 90 to 103 have been revised and decided to be kept in the literature review as they express arguments from important scientific research ( S. Poponi, A. Colantoni, S. R. S. Cividino, and E. M. Mosconi, “The Stakeholders’ perspective within the B corp certification for a circular approach,” Sustain., vol. 11, no. 6, pp. 1–15, 2019, doi: 10.3390/su11061584. / T. De Mendonca and Y. Zhou, “What does targeting ecological sustainability mean for company financial performance?,” Bus. Strateg. Environ., vol. 28, no. 8, pp. 1583–1593, 2019, doi: 10.1002/bse.2334. / C. Putnam Rankin and T. L. Matthews, “Patterns of B Corps Certification: The Role of Institutional, Economic, and Political Resources,” Societies, vol. 10, no. 3, p. 72, 2020, doi: 10.3390/soc10030072.). The literature review was enhanced with scientific information in the fields of sustainability and certification, and, B Lab certification and sustainability.	2	1	The authors could use a standard for a plural of index word, ‘indices’ (345) or ‘indexes’ (362).			su14095590_perova	0
+Factor analysis involves the examination of interdependence relationship, so it shouldn’t be expressed as a hypothesis that establishes dependence relationships. The economic dimension has been added in the former lines 180 and 181. We clarify that figure 1 is original and was created by the authors. Regarding the data, we confirm that it was taken from the official website, during the period between March and May 2021. The link provided (https://bcorporation.eu/directory) was the one used. In the meantime, the site has been changed and the directory of certified companies can be found in another link. Table 2 has been eliminated. To avoid the repetition of information in Table 2 and the paragraph after, we followed the recommendations of another reviewer by placing the paragraph as a footnote. The reduction in the number of companies was based on the analysis of the database and information collected from B Lab that allowed us to see that there was a change in the measurement model used in B Corp certification during the period January 2017 and March 2021. Thus, to ensure the 3 consistency of the database and the statistical analysis subsequently performed, we reduced the database to a recent period in which the same measurement model was used. Finally, the hypothesis was removed since, as suggested by another reviewer, it doesn't state what we are trying to test in this study.	2	1	If the rules of the Journal don’t allow to write down the Figures who is the author, I suggest to authors write it in a paragraph before the Fig 1, or in a footnote, according to Editor’s allowance.			su14095590_perova	0
+"We have replaced the word ""indicator"" with ""aspect"" to eliminate possible confusion with the 5 indicators in the B Impact Assessment Model (Figure 6). The text has been completely revised and the word 'indices' has been used as the standard. The word ‘chapter’ has been replaced by ‘section’. Finally, we are thankful for the good appreciation to the many models studied in our research."	2	1	The Cronbach’s Alpha ‘selected’ just ‘environment’ area of the B Lab model.			su14095590_perova	0
+The topic is “B Corp certification”. The text has been revised and the source has been added (E. Diez-Busto, L. Sanchez-Ruiz, and A. Fernandez-Laviada, “The b corp movement: A systematic literature review,” Sustain., vol. 13, no. 5, pp. 1–17, 2021, doi: 10.3390/su13052508.). Hypothesis 1 has been removed with the justification provided in response 5. Information was added on Cronbach’s Alpha and its impact on the need for B Impact Assessment improvement.	2	1	What is the main result of the B assessment with the confirmatory analysis?			su14095590_perova	0
+Thank you for the suggestions. The word was changed to “objective”. The discussion section has been improved in line with the suggestions made in the conclusion.	2	1	The authors presented and analyzed the variables/areas of certification with many models, it is interesting and is a basis for analysis in the next section.			su14095590_perova	0
+We added more coherent theoretical arguments to the discussion based on the most recently published relevant academic papers on the topic (lines 125-133, 153-158, 442-452). The incorporated studies include Yarram and Adapa (2021), Boukattaya and Omri (2021), Shakil (2021), Wang, Wilson & Li, (2021), Cordeiro et al 2020, Birindelli, et al. (2019), and Nuber & Velte (2021). In total, the list of references increased from 21 to 36. The details of these studies and the logic behind their inclusion are explained in the text (pages 3 and 4). For the quantitative papers, our guiding line is the inclusion of papers proposing a reliable strategy to address the problem of endogeneity characterizing virtually all studies dealing with gender diversity. In a nutshell, gender diversity and firms’ performances (be they financial, ESG or environmental) have reverse causality. More gender-diverse boards are found to improve firms’ performances, and at the same time, better performing firms are more attractive to females. Any study incorporating these two variables would suffer from endogeneity. Therefore, inference and results are biased. Finding an instrument for gender diversity becomes a concern of first importance. The task is challenging as all potential candidates for being a suitable instrument should be already part of the regression equation. So far, the literature has not found an instrument unanimously agreed upon. This is the reason for which our selection of papers is rather limited. We focused on those papers with a choice of instrumental variables that are well explained and papers that ‘’go around’’ the problem by making use of a method that does not rely on external instruments. This includes papers using difference-GMM or system-GMM estimators. Their principal advantage is the use of internal instruments instead of external ones. Indeed, the instruments chosen are difference and the level of the covariates themselves. There are only a few of these studies available, including ours. This again explains the relatively restricted number of references we are considering.	2	1	"3- Please reconsider the use of adjectives such as ""complicated"" (line 79), interestingly (line 88) and so on."			su14127346_makarova	0
+Initially, we opted for integrating the theoretical background disseminated throughout the text to connect to the findings of related papers and to avoid redundancy and repetition. Changes that we incorporated in response to this comment improve the overall theoretical framework of the paper, including concepts, definitions and links to relevant literature. We also articulated theoretical assumptions and added a comprehensive discussion of the observed phenomena. Specifically, we added critical mass theory discussion in the literature review section (lines 125-132). We added interpretation of corresponding results in the findings and discussions section (lines 445-452) and the conclusion section (lines 482-503). These changes in addition to the existing discussions of theories and concepts significantly improve the theoretical background of the paper.	2	1	Below we provide the details on how your comments and suggestions have been addressed.			su14127346_makarova	0
+We also emphasize the policy implications of these findings (lines 453-462, 497-503). To better reflect the policy implications of the study, we incorporate changes that are diffuse through the text. In particular, we argue that promoting gender diversity cannot deliver its full potential, and may even lead to adverse results unless it is accompanied by measures that mitigate political and economic uncertainty. These measures would create an enabling environment both for households and entrepreneurs. Such measures should increase agents’ confidence, dump their willingness to invest and expand the activity by reducing their aversion to risk. The accompanying measures should primarily target building and strengthening institutions to unlock this potential. Institutions are a defence against uncertainty. They contribute to anchoring economic agents’ expectations and stabilize the economy. We also elaborate on the finding of the critical mass theory. This result suggests that a very effective way to improve environmental quality and mitigate the adverse effects of economic activities is to encourage gender diversity in corporate boards in countries with low uncertainty where we find a higher impact on firms’ environmental performances. In these countries, the margin for progress is large and the effect of improving gender diversity in corporate boards materializes more easily and at lower costs, as these countries are far from their efficiency frontiers (lines 445-461 and 482-503).	2	1	We deeply regret the inconvenience caused due to this.			su14127346_makarova	0
+We addressed your suggestion to improve arguments and discussion of findings by revising existing and adding new paragraphs better explaining regression coefficients that demonstrate how political and economic uncertainty shapes the relationship between corporate board gender diversity and firms’ environmental performance. (Lines 379-389). Furthermore, we added supporting arguments disseminated across the text (particularly in lines 348-361, lines 379-389, etc). These relate our finding to theory. We focus here on the attitude towards risk and uncertainties. Women on corporate boards tend to be more risk-averse and lean more toward conservatism in uncertain environments. Therefore, the more females there are on corporate boards, the higher their weight in the board decisions would be, and by the same token, the higher the effect of uncertainty on firms’ decisions. Moreover, we also relate our findings to the critical mass theory, suggesting that the impact of gender diversity on the environmental performances of firms materializes only when a critical mass weight of females is realized in corporate boards (Konrad et al. 2008, Torchia et al. 2011, Schwartz-Ziv, 2017- page 12, lines 445-451).	2	1	They contribute to anchoring economic agents’ expectations, and stabilize the economy.			su14127346_makarova	0
+Absolutely. Changed accordingly (line 160).	2	1	The rational for this result stems from the idea that uncertainty adversely affects the enabling environment of an economy.			su14127346_makarova	0
+We answered this comment along different dimensions. We revise and elaborate on the explanation and interpretation of the results. We provide further explanations on the ways political and economic uncertainty shapes the relationship between corporate board gender diversity and firms’ environmental performance. We also emphasize the policy implications of these findings (lines 453-461, 487-496). In addition, we add more theoretical arguments to the discussion based on the most recently published academic papers on the topic. (Lines 421-452) “	2	1	Hi Lasha, We have investigated your query for RIC – OBE.TO for the year 2009(FY-12) Property/Plant/Equipment, Total - Net is bigger than Total assets, as reported and confirm that we have amended Property, plant and equipment which was incorrectly captured and sent the correct data to product.			su14127346_makarova	0
+The related theory posits that females’ attitude when facing uncertainty is different from males. They tend to exhibit a strong inclination to wait until the uncertainty is resolved before making major business decisions. To address this comment, we added supporting arguments disseminated across the text (particularly in lines 348-361, lines 421-428, etc). These explain the finding you are referring to by attitude towards risk and uncertainties. Women on corporate boards tend to be more risk-averse and lean more toward conservatism in an uncertain environment. Therefore, the more females there are on corporate boards, the higher would be their weight in the board decisions, and by the same token, the higher would be the effect of uncertainty on firms’ decisions.	2	1	More gender-diverse corporate boards would not lead to better environmental performances of firms in countries with higher uncertainty (higher WUI score).			su14127346_makarova	0
+The literature on the relationship between gender diversity and environmental performances of firms that addresses dynamic endogeneity is rather limited. We mention Gaio and Gonçalves (2022), Lu and Herremans (2019), Kassini et al (2016), Silla, et al (2016) that opt for a dynamic modelling using a System-GMM estimator. This estimator is particularly suitable to panels with large number of individual and few time periods, which the case for our paper. Moreover, it accommodates situations where the independent variable changes slowly (Antoniou et al., 2008). To keep a good flow in the text, we only mention these papers in the introduction section. We elaborate further on them in the literature review section. We describe their main purpose and explain how our analysis departs from them.	2	1	Similar problems stem from the following paragraphs discussing the results.			su14127346_makarova	0
+Thanks for the Suggestion. We highlighted the research hypothesis by moving it the introduction. We also added another hypothesis. The first relates to the very existence of a relationship between gender diversity on corporate boards and firms’ performances. The second, deals with the intensity and shape of this relationship. We believe, this adds a deal of clarity to the text and purpose of the paper.	2	1	Your review truly helped improve further the paper.			su14127346_makarova	0
+We reviewed all the text and changed wording in different parts of the text.	2	1	5- Please revise your data: your max tangibility is over 2 (which means that Net PP&E, a subset of Assets, is 2 times those same Assets - this should be a typo.			su14127346_makarova	0
+We initially meant, among all these papers, there is one that… We rephrased this sentence and reviewed all the text. We edited sentences when needed.	2	1	I see the work interesting, although it needs a good reorientation, in order to make it more attractive to reading.			su14127346_makarova	0
+In our initial submission, we used winsorized data for all our econometric analysis. However, we, inadvertently, displayed summary statistics (Table 2) for raw data, i.e., before winsorizing. We should have reported only winsorized summary statistics. This is precisely what we do in this revised version of the paper. The reason for winsorizing data (beyond its advantage of having regular statistical distribution) was that we initially have detected outliers. Further investigation revealed that one company (RIC: OBE.TO; CompName: Obsidian Energy Ltd) is responsible for these outliers. The company had presumably reported wrong values in FY-12, where PP&E was higher than Total Assets (see table below from Refinitiv Eikon). We have reported this case to Refinitiv (Case number: 11297339) and our doubts were confirmed (See their response below). Refinitiv Eikon replaced old value of PP&E (27834189009) with a correct value (10788172656). In order to avoid similar mistakes, we opted for winsorizing data and avoid outliers. Similarly, only one company had negative Tobins’ Q in 2017 (RIC: ELR.TO ; company name: Eastern Platinum Ltd), which was a reporting typo as well. However, our analysis is not affected by these outliers which are removed after winsorizing all the variables. In this revised version of the paper, we replace the table of summary statistics with the one based on winsorized variables.	2	1	Gender diversity on the board and firms’ corporate social responsibility.			su14127346_makarova	0
+The sentence “The global average of corporate boards’ gender diversity for all the sectors during 2008-2018 was about 13.5%” is correct as it is calculated based on all industries (5120 firms in total). However, since we are focusing on “Mining, Quarrying, and Oil and Gas Extraction Industry”, it makes more sense indeed to report the value for that particular sector. Therefore, we update the text as follows: “The average of corporate boards’ gender diversity for Mining, Quarrying, and Oil and Gas Extraction Industry during 2008-2018 was about 9.6%”.	2	1	Overall, I believe that this study has a high potential to add to the literature on the relationship between gender diversity and firm environmental performance.			su14127346_makarova	0
+This is an explanation of the following sentence in our article: “To avoid bias and inconsistency that might be caused by instruments proliferation, we collapse the instruments matrix.” We are using longer lags of the dependent variable as additional instruments. Intuitively, there is a trade-off, on one hand, using longer lags gives more information and improves efficiency, but on the other hand sample gets smaller (as it is the case in standard 2SLS models). GMM is a way around this trade-off between lag length and sample length. However, implementation of System-GMM is not exempt from weaknesses. With the instruments count being quadratic in the time dimension, the variance matrix of the moments can be very large. Estimating its elements may be problematic in particular in a finite sample lacking the adequate amount of information. This finite sample bias can weaken Hansen J-test and overstate its p-value and therefore generate suspicious high pass rate of instruments validity. The problem is known as instruments proliferation (Roodman 2009). To avoid bias and inconsistency due to instruments proliferation, we limit the number of instruments used and collapse the instrument matrix which prevents the model from being over-fitted.	2	1	I see the work interesting, although it needs a good reorientation, in order to make it more attractive to reading.			su14127346_makarova	0
+We revised the concerned section and rewrote the text in order to improve on clarity. The revised text reads as follows: Greater gender diversity for a firm located in a country with high political and economic uncertainty would lower the firm’s environmental performance (0.616-0.853=-0.237).	2	1	"2 4- What do you mean by ""in the crowd"" (line 141)."			su14127346_makarova	0
+the inoculation points were labeled with the red arrows in Figure 1 as suggested.	2	1	The following are some comments and suggestions.			toxins14060409_makarova	0
+Sorry for carelessness. We corrected it as suggested. (Pag.4 Line 157) Point 3: Line 143: “PPSD-premature spike death” should be “PPSD-proportion of premature spike death”.	2	1	Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			toxins14060409_makarova	0
+We revised the expression in the footnote of Table 1. (Pag.4 Line 158) Point 4: Lines 168 - 169: Should it be that DON content of most lines without Fhb1 exceeded 1000 µg/kg under UBFI?	2	1	While DON concentrations in grains are usually correlated with disease severity, high DON levels have been observed in asymptomatic kernels.			toxins14060409_makarova	0
+we revised the sentence as “DON content of most lines without Fhb1 exceeded 1000 μg∙kg-1 under UBFI method (Figure 2), with the highest reaching 4224.38 μg∙kg-1 (Table S1)”. (Pag.5 Line 184-186) Point 5: Lines 411 & 413: “PPSB” should be “PPSD”.	2	1	The paper describes a set of experiments carried out in 2020 2021 on isogenic lines of wheat using different methods of Fusarium inoculation in order to assess the effect of Fusarium inoculation on FHB disease index and DON accumulation.			toxins14060409_makarova	0
+We corrected it. (Pag.11 Line 441 & 443) Point 6: Table S1: “PPSD - premature spike death” in the footnote should be “PPSD - proportion of premature spike death”.	2	1	Among the three methods used, the upper bilateral floret injection (UBFI) maximizes DON accumulation in grain and significantly distinguishes DON content in different wheat genotypes, indicating that the method could be useful for breeding program to develop wheat cultivars with resistance to DON accumulation.			toxins14060409_makarova	0
+We revised the expression in the footnote of Table S1.	2	1	1 Response to Reviewer 2 Comments Point 1: I would suggest to avoid citing supplementary figures at the beginning of the chapter.			toxins14060409_makarova	0
+Thanks. We revised the annotations in Table S1.	2	1	The paper is of high interest for the community working on FHB and shows the importance of inoculation method on the results of disease and mycotoxin accumulation.			toxins14060409_makarova	0
+We agree with the reviewer’s comments and make Figure S4 as Figure 4 in this revised version.	2	1	Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			toxins14060409_makarova	0
+We marked the inoculation sites with red arrows in Figure 1.	2	1	Point 7: Fhb1- and Fhb1+ don’t appear in Table S1, so they can be removed from the footnote.			toxins14060409_makarova	0
+We added more details about the strain and cited the following paper. (Pag.10 Line 402-403) Zhang, Y.; Li, A.; Zhu, S.; Li, L.; He, X.; Sun, Z.; Li, T. Basal Rachis Internode Injection (BRII): A novel inoculation method to evaluate wheat resistance to Fusarium head blight. Phytopathology, 2021, 111, 1670-1674, doi:10.1094/PHYTO-11-20-0488-R. Point 4: Mycotoxin data analysis should be provided (I guess is policy of mdpi to make raw data available together with the publication) Response 4: Thanks!	2	1	Mycotoxin deoxynivalenol (DON) contamination of wheat and other grains has been a concern of food safety.			toxins14060409_makarova	0
+Thanks! We uploaded the raw mycotoxin data as suggested.	2	1	Given that 2.4 chapter discusses data of supplementary figure 4 would suggest to make figure S4 as Figure 4 of the manuscript.			toxins14060409_makarova	0
+Following the suggestion of this reviewer, we have removed the movie in the revised version.	2	1	Figure 2B shows that the amount of IFITM3 is not significantly altered up to 6 h p.i.			v11060548_makarova	0
+We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.	2	1	(Fig. S2B), as was previously described by others.			v11060548_makarova	0
+The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.”	2	1	There are few clarifications that would strengthen the overall manuscript.			v11060548_makarova	0
+We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.	2	1	The supplemental figures only include experiments that reproduce previously published work or are not absolutely necessary to support the major and most novel findings of the paper.			v11060548_makarova	0
+We have made the suggested changes.	2	1	The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			v11060548_makarova	0
+We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.	2	1	This study provides imaging of the antiviral protein IFITM3 at endogenous levels and at super-resolution in the context of influenza virus infection.			v11060548_makarova	0
+To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.	2	1	There are few clarifications that would strengthen the overall manuscript.			v11060548_makarova	0
+If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.	2	1	Figure 4: Label the figure so that it can be interpreted independent of the legend.			v11060548_makarova	0
+We have made the requested changes.	2	1	In this study the authors examine the localization of IFITM3 during a viral infection in airway epithelial cells.			v11060548_makarova	0
+We have made the requested changes to figure 5A/C and Figure 7.	2	1	It seems (as the authors state) that these represent abortive events (Figure 7).			v11060548_makarova	0
+We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).	2	1	It seems (as the authors state) that these represent abortive events (Figure 7).			v11060548_makarova	0
+Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)	2	1	Figure 4: Uninfected control should be shown in the main text.			v11060548_makarova	0
+All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.	2	1	This may be true, but far more studies have confirmed this link than have refuted it.			v11060548_makarova	0
+Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.	2	1	Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu.			v11060548_makarova	0
+"We thank the reviewer for pointing out the inaccuracy in the references. The statement ""...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection"" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3."	2	1	If this is not acceptable we will make the changes in final revision.			v11060548_makarova	0
+The supplemental figures only include experiments that reproduce previously published work or are not absolutely necessary to support the major and most novel findings of the paper. We therefore suggest to keep these data in the supplement.	2	1	The authors have largely addressed my previous comments.			v11060548_perova	0
+We show indirect immunofluorescence analysis with anti-IFITM3 and anti-NP antibodies of HSAEpCs in Figure S6. No unspecific signal is detected in the cytosol of HSAEpCs, indicating that the observed cytoplasmic NP stain is specific. Nuclear NP stain is detected at 10 h p.i., and this may indeed indicate delayed replication compared to A549 cells as suggested by the reviewer, but no direct side-by-side comparison of replication kinetics in different cells was performed.	2	1	There are few clarifications that would strengthen the overall manuscript.			v11060548_perova	0
+The IFITM3 images were acquired in the cytosol distant from the nucleus and absent from the plasma membrane. We included this information in the respective figure legend of Figure 3 and 7. Figure 3: “STED images (raw data) show representative subcellular regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane as used for cluster analysis.” Figure 7: “Images show regions located in the cytosolic part distant from the nucleus and absent from the plasma membrane.” d.	2	1	Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			v11060548_perova	0
+We now include the Western Blot analysis suggested by the reviewer as Figure S2B and thank the reviewer for suggesting this important control.	2	1	Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.			v11060548_perova	0
+We have made the suggested changes.	2	1	While there are important new insights provided by this work, the manuscript can be improved in terms of data presentation and referencing the existing literature on the IFITM proteins.			v11060548_perova	0
+We now include an immunoblot showing IFITM3 levels in whole-cell lysates from A549 wildtype and IFITM3 knock-down cells as Figure S1D.	2	1	Figure 7: I am not sure that this figure is a useful addition to the paper as it suffers from exactly the problems that the authors claim to overcome in their study, i.e., the use of overexpressed IFITM3 and an overexpressed Rab11 marker protein, both of which may show unnatural localization due to the overexpression.			v11060548_perova	0
+To address the reviewer’s concern, we performed statistical analysis of three independent experiments shown in Figure 2B, lower panel. Statistical analysis of mean values of the determined protein amount for IFITM3 shows no statistically significant differences (e.g. p-value of 6 h pi compared to control is 0.5224). We included this information in the figure legend of Figure 2.	2	1	Figure 3A: Why is there so much IFITM3 present in the uninfected cells?			v11060548_perova	0
+Figure 2B shows that the amount of IFITM3 is not significantly altered up to 6 h p.i. IFITM3 protein increased significantly after longer infection times such as 24 h p.i. (Fig. S2B), as was previously described by others.	2	1	Supplemental figures should be included in the main text.			v11060548_perova	0
+If this is not acceptable we will make the changes in final revision. We have not made this change as we show that the anti-NP antibody is highly specific and shows negligible signal in the uninfected controls in Figure 1A. A representative example of non-infected HSAEpC is shown in the supplement.	2	1	Figure 4: Label the figure so that it can be interpreted independent of the legend.			v11060548_perova	0
+We have made the requested changes.	2	1	The authors have largely addressed my previous comments.			v11060548_perova	0
+We have made the requested changes to figure 5A/C and Figure 7.	2	1	It seems (as the authors state) that these represent abortive events (Figure 7).			v11060548_perova	0
+We used a fluorescently tagged Rab11 because the anti-Rab11 antibodies were not sufficient to label recycling endosomes. However, IFITM3 is detected at native levels and this compromise allowed us to evaluate its localization. Overexpressed IFITM3 was only used for the live cell microscopy shown in the movie, which has now been removed (see reply to reviewer 1).	2	1	Figure 4: Label the figure so that it can be interpreted independent of the legend.			v11060548_perova	0
+Based on the reviewer´s suggestion, we have now included again the reference showing the infection increase of IAV in A549 IFITM3 knockdown cells and have changed the wording to make the statement clearer. “The mechanism by whichIFITM3 impedes IAV infection was divergent in previous studies and inhibition was mostly described upon IFITM3 over-expression [37, 38, 51]. It was demonstrated that IFITM3 knockdown in A549 cells increases infection rate [37].” Since this paper is primarily studying IFITM localization and clustering at endosomes/lysosomes it should reference studies in which endocytic localization motifs of the IFITMs were identified and characterized (Jia, Cell Microbiol, 2014; Chesarino, JBC, 2015; Li, JBC, 2015)	2	1	Using super-resolution microscopy the authors show colocalization of IFITM3 with influenza viral nucleoprotein and early endosomal proteins.			v11060548_perova	0
+All the references have been added and we now include this aspect of the IFITM3 mechanism in the discussion of the paper. We thank the reviewer for the comment.	2	1	The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			v11060548_perova	0
+Lines 61 ff. were changed pointing out the relevance of SNP rs12252-C and SNP rs34481144-A. References have been added.	2	1	Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017).			v11060548_perova	0
+"We thank the reviewer for pointing out the inaccuracy in the references. The statement ""...IFITM3 elevates the level of cholesterol on late endosomes and lysosomes thereby restricting early IAV infection"" is based on the publication of Kühnl et al. 2018 (mBio). The reference was replaced. We have also complemented the introductory part with the aspect of fusion inhibition through the amphipathic helix of IFITM3."	2	1	If this is not acceptable we will make the changes in final revision.			v11060548_perova	0
+The innovation of this research has been clarified in the abstract, the introduction, and the conclusion.	2	1	Authors should add a list of abbreviations before the references.			w14030367_perova	0
+Three references have been cited. Please see lines 40-41 in the revised manuscript.	2	1	The article topic is intriguing and promising in the area.			w14030367_perova	0
+Thanks very much for the reviewer. The conclusion has been revised according to the reviewer’s suggestion. The added paragraph in the revised manuscript is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conventional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.	2	1	Response 10: Thanks very much for this comment.			w14030367_perova	0
+Thanks for the reviewer’s suggestion. The conclusion has been revised and integrated into two paragraphs. The first paragraph mainly includes important findings, and the second paragraph mainly includes the outlook for the future in this field.	2	1	Point 5: The novelty of the work must be clearly addressed and discussed, compare previous research with existing research findings and highlight novelty.			w14030367_perova	0
+The salinity concentrations in the samples have been added to Table 1 in the revised manuscript.	2	1	However, these results are not shown in the manuscript, how many replicated were carried out by experiment?			w14030367_perova	0
+We agree with the suggestion and comments of the reviewer. All comments have been revised one by one, and the modifications are highlighted in yellow in the marked revised manuscript.	2	1	Study and evaluation of the characteristics of saline wastewater (brine) produced by desalination and industrial plants.			w14030367_perova	0
+Thanks for the reviewer’s helpful suggestion. The objective statement has been added to the abstract to support the seriousness of the problem, and the revised sentences are as follows: Biological activated carbon (BAC) biofilter coupling ultrafiltration (UF) is a promising process for the treatment of river water contaminated by pharmaceutical and personal care products (PPCPs). However, the pilot-scale study should be conducted to reveal the long-term removal performance and the respective contributions of BAC and UF.	2	1	43 Citation: Wanga, Q.; Tanga, X.; Zenga, W.; Wangb, F.; Gongb, W.; Chena, J.; Wanga, J.; Lia, G.; Lianga, H. Pilot-scale biological activated carbon filtration-ultrafiltration (BAC-UF) system for removing pharmaceutical and personal care products (PPCPs) from surface wa- ter.			w14030367_perova	0
+We agree with the suggestion of this reviewer. The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances. However, the mechanisms of the two-stage biofilm, such as bacterial and metazoan communities, membrane fouling and dissolved oxygen transfer, should be further investigated to enhance the removal efficiency and stability of this system. The research gap has been added in the Conclusion and is as follow: To sum up, the results show that the proposed BAC-UF system can be effective in the treatment of river water polluted by PPCPs, conventional organic pollutants and ammonia nitrogen. Besides, the results of this analysis can have significant implications for the conven-tional UF operation procedure and the ozone-activated carbon process, providing a simple decentralized approach to drinking water treatment for the areas where source water is contaminated with PPCPs.	2	1	Point 3: Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10].			w14030367_perova	0
+Thanks for the reviewer’s comment. The introduction has been revised carefully, including using up-to-date references. For example, the outdated references have been removed. Meanwhile, the articles published within the last two years were added, such as (the Reference2 Yu et al., 2020) and (the Reference21 Tang et al., 2018).	2	1	Nev- 21 ertheless, BAC and UF were both related to the removal effects of NH4+-N, NO2−-N and NO3−-N. 22 Importantly, the BAC-UF process showed high efficiency in the removal PPCPs micropollutants 23 and the average removal of total PPCPs reached 47.84%, especially Anhydroerythromycin, Sul- 24 fachloropyridazine, Sulfadiazine, Trimethoprim, and Caffeine.			w14030367_perova	0
+The reference has been cited. Please see line 38 in the revised manuscript.	2	1	Point 15: Please add a comparative profile section to compare your results and prove how it better than previous.			w14030367_perova	0
+The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water. The above coupling process makes up for the defects concerning respective operations of the BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the different potential of biofilm growing. As far as we investigated, the lack of enough attention to long-term pilot study exists. Therefore, in this study, a BAC-UF system was carried out for several months with pilot scale to access the long-term removal performances and the respective contributions of BAC and UF. The relevant content has been supplemented and revised in the abstract to highlight the innovative points.	2	1	Please revise your paper accordingly since several language issue occurs on several spots in the paper.			w14030367_perova	0
+Biological activated carbon (BAC) combined the adsorption and biologic degradation consuming low power energy and chemicals without concern of DBPs production as well as no frequent updates for activated carbon media. The BAC biofilter can remove the PPCPs pollutants, and then the followed UF can reject microorganisms and particles flowing out from the biofilter to ensure the quality of drinking water. Thus, the BAC-UF process makes up for the defects of the respective operations of BAC biofilter and UF. Although many pilot-scale setups were used to treat secondary wastewater effluent for water reclamation, this type of raw water quality was different from the river water, causing the potential of biofilm growing differently. As far as we investigated, the lack of enough attention to long-term pilot study is present. Therefore, in this study, a BAC-UF system was carried out for several months with a pilot scale.	2	1	In this study, a 17 BAC-UF system with treatment capacity of 0.16 m3/h was operated.			w14030367_perova	0
+The two references have been cited. Please see the lines 58.	2	1	However, the mechanisms of the two-stage biofilm, such as bacterial and metazoan communities, membrane fouling and dissolved oxygen transfer, should be further investigated to enhance the removal efficiency and stability of this system.			w14030367_perova	0
+The introduction has been improved for better reading.	2	1	Response 6: Thanks very much for this comment.			w14030367_perova	0
+Two PPCP mistakes have been revised. The abbreviation of chemical oxygen demand in previous version has been corrected to CODMn. The unnecessary abbreviations have been also corrected in the abstract.	2	1	Hope the revised manuscript would be more satisfactory.			w14030367_perova	0
+The added paragraph is as follows: Potassium permanganate, H2SO4, potassium sodium tartrate tetrahydrate, Nessler’s reagent and NaOH were purchased from a commercial company and certified as AR purity (Guangzhou Chemical Reagent Factory, Guangzhou, China), while PPCP standards were provided by three companies. Sulfamethoxazole (SMX), Sulfadoxine (SD), Sulfachloropyridazine (SCP), Sulfamethoxypyridazine (SMP), Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Erythromycin (EM), Anhydroerythromycin (EA), Roxithromycin (ROX), Penicillin-G (PEN G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacine (IND), Metoprolol (METO), Propranolol (), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), and Sulpiride (SP) were obtained from the Dr. Ehrenstorfer Company in Germany. Furthermore, Sulfadimidine (SM2), Sulfadiazine (SDZ), Sulfapyridine (SP), Sulfamonomethoxine (SMM), Tetracycline (TC), Ofloxacin (OFL), Amoxicillin (AM), Dimetridazole (DMZ), Trimethoprim (TMP) were bought from the National Institute of Metrology in China, whereas Oxytetracycline (OTC), Caffeine (CF), and Diethyltoluamide (DEET) were acquired from the Toronto Research Chemicals Company in Canada.	2	1	Point 15: Please add a comparative profile section to compare your results and prove how it better than previous.			w14030367_perova	0
+These three GAC-UF systems were operated in parallel. Samples of the feedwater and effluent from three systems were taken simultaneously and measured once. The standard deviation was obtained by the detection over the full period of the experiment.	2	1	The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water.			w14030367_perova	0
+The figure is used as the only pattern in the revised manuscript.	2	1	Response 3: Thank you very much for your kind reminder.			w14030367_perova	0
+The front sizes in the figure 1 have been increased. The quality of the figure 1 has been also improved.	2	1	They concluded based on the results from this study that the BAC-UF process is a promising process for producing clean drink water.			w14030367_perova	0
+The style for units has been revised and unified.	2	1	Besides, the t-test was used and proved the significant difference between BAC-Effluent and BAC/UF-effluent.			w14030367_perova	0
+The comparison of previous research with existing research findings was added in the final part of the results and discussion. The added paragraph is as follows: PPCPs risks have been posing severe challenges to the safety of drinking water supply in rural areas due to the absence of the process with simple operation and maintenance as well as reliable performance. In this study, BAC coupling gravi-ty-driven UF was performed continuously, and the rejection performance of mem-brane filtration and BAC filtration both showed barriers for the conventional pollu-tants and PPCPs. Furthermore, this study indicated the respective contributions of BAC and UF, showing the role of the two-stage biofilm. Previous works involving BAC generally combined the ozonation with the BAC filter for treating the contaminants of emerging concern, eliminating a majority of PPCPs by more than 90% [34]. However, the regulation and maintenance of machines for ozone products are complicated, and the disinfection by-products will be newly generated in the effluent, which is incon-venient to use in rural areas [35]. In general, coagulation, filtration and single BAC units worked inefficiently and removed the detected PPCPs by less than 50%, as they were not hydrophobic [34, 36]. Hybrid membrane processes such as inline dosing of powdered activated carbon (PAC) prior to UF have already shown promising potential for the abatement of PPCPs; however, the inline dosing PAC is infeasible in rural areas [37]. In this study, the BAC prior to UF enhanced the biological activity by forming a two-stage biofilm system. Therefore, the integrated BAC-UF process can be considered as an economically and technically feasible approach to the decentralized and emer-gency drinking water treatment.	2	1	Response 3: Thank you very much for your kind reminder.			w14030367_perova	0
+The conclusion has been modified as suggested by reviewers. Please see lines 325-350 in the revised manuscript.	2	1	Point 6: I strongly suggest that the authors explain how they performed the t-tests.			w14030367_perova	0
+Two native English-speaking colleagues help us verify the manuscript. Hope the revised manuscript would be more satisfactory. We are so sorry to make reviewer’s reading uncomfortable. We have used an English Language Editing service to correct the grammatical and spelling errors and to make the expressions conform to correct scientific English (the Language Editing Certification is attached below).	2	1	Response to Reviewer 4 Comments Point 1: Authors should avoid abbreviations in the title and the abstract.			w14030367_perova	0
+We agree with the reviewer. The reference formatting has been corrected. Thanks very much for the patient review again.	2	1	Response 10: Thanks very much for this comment.			w14030367_perova	0
+Full names of BAC, UF, and PPCPs are used in the title and the abstract to show the results briefly. As for NH4+-N, NO2−-N, and NO3—N, the full names have been used instead of abbreviations in the abstract.	2	1	Response 7: Thank you very much for your kind reminder.			w14030367_perova	0
+The title has been revised to river water.	2	1	Please target the specific gap such as 2015-2021 etc Response 3:			w14030367_perova	0
+The references format has been corrected.	2	1	The PPCPs catalog includes a variety of chemicals, such as human and veteri- 36 nary drugs, fragrances and disinfectants used in personal care products (e.g., soaps, lo- 37 tions, sun-screens and body cleaning products) and household chemicals [3].			w14030367_perova	0
+We are very sorry for the confusion caused to the reviewers, the sentences have been revised from the words and Grammarly. The revised sentences are shown below: Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, col-loids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10]. However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite [11, 12].	2	1	Point 15: Please add a comparative profile section to compare your results and prove how it better than previous.			w14030367_perova	0
+The t-test results have been added in the revised Table1.	2	1	Title: Pilot-scale biological activated carbon filtration-ultrafiltration (BAC-UF) system for removing pharmaceutical and personal care products (PPCPs) from surface water.			w14030367_perova	0
+"The statistical t-test was evaluated for Figure 4, which was added in the text of ""3.2. Removal of nitrogen"" Section."	2	1	Response to Reviewer 3 Comments Point 1: The title seems good, but the abstract seems to be fine.			w14030367_perova	0
+The t-test results have been added to the revised Table2. The concentration of Erythromycin was generally varied between 1047.14 ng L−1 and 2037.72 ng L−1.	2	1	Point 9: Point 6: Why the same trend of increase or decrease in graphs (Fig.			w14030367_perova	0
+Some symbols were overlapped in Figure 5h. The symbols near 85 days have been entirely shown by correcting Y-axis settings, such as Figures 5a and 5f.	2	1	Point 4: Line 44-45: Ultrafiltration (UF) as emerging technology, has been widely used to remove pollutants such as particles, colloids, bacteria and viruses, reducing the risk of water-borne diseases and …UF membranes cannot effectively rejected these soluble substances”.			w14030367_perova	0
+The conclusion has been revised, including the style and sentences.	2	1	As far as we investigated, the lack of enough attention to long-term pilot study exists.			w14030367_perova	0
+We agree with this suggestion. The abbreviations in the title have been deleted. Besides, a list of abbreviations has been added before the references.	4	1	It was found that the BAC unit 25 played a key role in PPCPs removal and the UF unit also degraded them by the biomass on UF 26 membranes.			w14030367_perova	0
+Thanks to the reviewer for the patient comment. The above mistakes have been revised.	4	1	Point 6: I strongly suggest that the authors explain how they performed the t-tests.			w14030367_perova	0
+1. 2. Thanks to this reviewer for the comment. Common waterborne viruses include Shigella, Vibrio cholerae, Poliovirus, adenovirus, and coxsackie virus, with sizes of 26 nm, 27 nm, 30 nm, 90 nm, and 30 nm, respectively [1-5]. The molecular weights of PPCPs are generally less than 1000 Da (about 1-2 nm), obviously smaller than the virus. The molecular weight cut off of UF membranes was an average 100 000 Da in this work, which was similar to the virus but larger than the PPCPs molecules. Karthik K, Dhanuskodi S, Gobinath C, et al. Multifunctional properties of microwave assisted CdO–NiO–ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities [J]. Journal of Materials Science: Materials in Electronics, 2018, 29(7): 5459-71. Song S, Liu Z, Zhou J, et al. An adjuvant compound that enhances immunogenicity at fractional doses of the Sabin-inactivated poliovirus vaccine (sIPV) with a long duration of protection in a rat model [J]. Journal of Medical Virology, 2019, 91(1): 14-21. Kim K, Choi J-W, Ma K, et al. Nanoisland-Based Random Activation of Fluorescence for Visualizing Endocytotic Internalization of Adenovirus [J]. Small, 2010, 6(12): 1293-9. Dourmashkin R R, Mccall S A, Dourmashkin N, et al. Virus-like particles and enterovirus antigen found in the brainstem neurons of Parkinson's disease [J]. F1000Res, 2018, 7: 302-. Tamano K, Aizawa S-I, Katayama E, et al. Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors [J]. The EMBO Journal, 2000, 19(15): 3876-87.	4	1	Authors may provide data in the supporting information file.			w14030367_perova	0
+The information regarding the biological degradation of PPCPs has been added in the introduction. The revised sentences are shown below: In the BAC biofilters, the biotransformation and adsorption both contributed to the PPCPs removal. The activated carbon adsorbed PPCPs to the surface and interior, where microorganisms were suitable for growth. Under the long-term effect of high-concentration PPCPs, the dominant microorganisms in the biofilter were selected to survive. These microorganisms mostly transformed PPCPs into many segments and even directly mineralized them to CO2 [3, 14].	4	1	Why is the DO concentration of BAC-effluent sometimes higher than the Raw water.			w14030367_perova	0
+“The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”. This claim is not supported by our research. To avoid misunderstandings for readers, we changed the claim and revised the sentence. The revised version is as follows: The combined process with UF may be another promising choice as an alternative to nanofiltration for removing PPCPs in rural areas. It can be seen in lines 54-55.	4	1	Point 13: Please provide high quality image of figure 1.			w14030367_perova	0
+The method of the t-tests has been explained in section 2.4 (lines 157-158) in the revised manuscript. The sample size (n) has also been added in Table 1 and Table 2.	4	1	Please cite this reference with existing reference 3….Role of nanotechnology for design and development of cosmeceutical: application in makeup and skin care.			w14030367_perova	0
+Thank you very much for your kind reminder. The values have been modified in the revised manuscript in yellow highlight.	4	1	Response 10: Thanks very much for this comment.			w14030367_perova	0
+Thanks for the kind reminder. All the names of medicinal compounds not used have been deleted, including Sulfaquinoxaline (SQX), Sulfathiazole (STZ), Doxycycline (DOX), Roxithromycin (ROX), Penicillin-G (PEN-G), Clarithromycin (CAM), Norfloxacin (NOR), Oxociprofloxacin (OFL), Enrofloxacin (EFL), Flumequine (FQ), Acetaminophen (APAP), Diclofenac sodium (DCF), Naproxen (NAP), Indomethacin (IND), Metoprolol (METO), Propranolol (PRO), Atenolol (ATL), Primidone (PRM), Carbamazepine (CMZ), Sulpiride (SP), Sulfapyridine (SPN), Sulfamonomethoxine (SMM), Tetracycline (TC), Amoxicillin (AM), Dimetridazole (DMZ), Oxytetracycline (OTC) and DEET.	4	1	Point 4: The authors should include some information regarding the biological degradation of PPCPs by microorganisms in the introduction section.			w14030367_perova	0
+The results and discussion has been supplemented in section 3.1. Please see lines 180-185 in the revised manuscript.	4	1	Response 12: Thanks very much for this comment.			w14030367_perova	0
+Thanks very much for this comment. With a turbulent current and dozens of kilometers in length, this river is located in the mountains of Foshan city. The PPCPs sludge at the bottom of the river never executes removal. Thanks for the suggestion from the reviewer. We will further precipitate the PPCPs sludge before the water treatment plant or excavate the PPCPs sludge from the river and study the effects on drinking water quality. The relevant results will be published in another paper in the future.	4	1	Response 7: Thank you very much for your kind reminder.			w14030367_perova	0
+Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.	2	1	The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar.			w14081258_makarova	0
+It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615. The values obtained of I DW and steric hindrance are not explained.	2	1	The degradation of these isomers were stable within the former several days, which mentioned in the manuscript.			w14081258_makarova	0
+The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications. Shiyu Wang, Fei Liu*, Wenyong Wu*, Yaqi Hu, Renkuan Liao, Gaoting Chen, Jiulong Wang, Jialin Li. Migration and health risks of nonylphenol and bisphenol A in soil-winter wheat systems with long-term reclaimed water irrigation. Ecotoxicology and Environmental Safety 158 (2018) 28–36. Shiyu Wang，Wenyong Wu*，Fei Liu, Xiaoou Li. Sorption and desorption behaviours of 4-nonylphenol on reclaimed water-irrigated soils. Environmental Engineering Science. (2019) 36 (9) : 1100-1111。 王世玉，刘菲*，刘玉龙，陈亮. 气相色谱-质谱法检测地下水中12种对壬基酚同分异构体.分析化学，2013，41（11）：1699-1703. 王世玉，刘菲*，吴文勇，尹世洋，刘玉龙，陈亮，张伟，陈会会.影响12种壬基酚同分异构体液液萃取效率的因素研究. 岩矿测试，2014，33（4）：570-577.	2	1	Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP.			w14081258_makarova	0
+It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.	2	1	Yours sincerely, Shiyu Wang [email protected] Author Response File: Author Response.docx			w14081258_makarova	0
+Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu. Spatial distribution and migration of nonylphenol in groundwater following long-term wastewater irrigation. Journal of Contaminant Hydrology,2015,177-178（June–July）:85-92. I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.	2	1	At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States.			w14081258_makarova	0
+It has been given in Table1.	2	1	Your careful review of this manuscript is highly appreciated.			w14081258_makarova	0
+Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.	2	1	The irrigation water physi-chemical parameters should be given.			w14081258_makarova	0
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.	2	1	Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.			w14081258_makarova	0
+Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.	4	1	Your careful review of this manuscript is highly appreciated.			w14081258_makarova	0
+Accepted and revised in L204. Figure 1 has been changed in the manuscript.	4	1	However, the degradation rate could be affected by this.			w14081258_makarova	0
+Accepted and revised in L95-105. Five-point sampling method was used in both fields (0-20cm). A total of 5 topsoil samples in each field were taken using a stainless steel spade and mixed thoroughly. Afterwards the soil samples were chosen by quartation; Then plants’ debris and residues were removed. Finally soil samples were 0.3-mm sieved and stored in brown glass bottles at 4 °C. The following was the area that the Florida soil was collected.	2	1	Please feel free to contact me if any questions.			w14081258_perova	0
+It has been mentioned in L163-164 in the manuscript. Randic (1975) proposed molecular branching index χ, hereinafter referred to as simple molecular connectivity index (1995). Therefore, they are the same concept. [38] Randic M., Razinger M. (1995). Molecular Topograhic Indices. Journal of Chemical Information and computer Sciences. 35: 140-147. [40] Randic M. (1975). On Characterization of molecular branching. Am. Chem. Soc. 97: 6609-6615.	2	1	In the future study, this would be taken into accounted.			w14081258_perova	0
+Accepted and revised in L157-160. Common topological indices, such as κ shape indices and information indices were also calculated by Molconn-Z (version 4.12S, eduSoft, La Jolla, CA). The detailed list and definitions can be found in the software user’s guide [35], and the definitions can also be found in Todeschini et al. [36]. The results of other common topological indices calculated by Molconn-Z were shown below as well. But in this study, only the steric index (R2 = 0.82 for CN soil; R2 = 0.86 for FN soil) and IDWbar (R2 = 0.83 for CN soil; R2 = 0.94 for FN soil) have a better relationship with the half lives of the isomers, which were mentioned in the Results and Discussion part. The results of the topological indices of the isomers [35] Hall, L. H.; Kellogg, G. E.; Haney, D. N. (2008). Software Package for Molecular Topology Analysis User’S Guide. [36] Todeschini, R.; Consonni, V.; Mannhold, R.; Kubinyi, H.; Folkers, G. (2009). Molecular Descriptors for Chemoinformatics; Wiley: New York.	2	1	If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate.			w14081258_perova	0
+The analytical methodology of isomers quantification has been studied and published in former study (Wang et al., 2013). From Figure1 in Supporting Information, it seems that the isomers were not separated, but actually it does not affect the quantification of the isomers. Because the main quantitative ions were used for quantification and the main quantitative ion of each isomer were separated absolutely (Figure 1). Table 1 and Figure 1 were cited from Wang et al (2013). Moreover, series of research about the NP isomers were conducted and published afterwards. The following are the publications.	2	1	Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu.			w14081258_perova	0
+It has been given in Supporting Information 2. Indeed, the characteristics of the soil are too limited. My visiting time was only one year. At the end of the visiting time, I had no time to complete the microbiological test and the soil properties detection and was going to leave the United States. So I had to acquire the Florida soil properties information by website. However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.	2	1	Figure 1 legends of axis need font size larger, please make readable the figures R：Accepted and revised in L204.			w14081258_perova	0
+I think it is a little difficult for me to acquire the irrigation water physi-chemical parameters of Florida, but I can acquire the NP load in China irrigation water, which were cited from my former study of Wang et al., (2015). Bu in this study, the irrigation water was prepared in Lab according to the max NP concentration in reclaimed water in the study area to simulate NP in reclaimed water in the actual environment.	2	1	Indeed, the characteristics of the soil are too limited.			w14081258_perova	0
+It has been given in Table1.	2	1	Worse more, the soil can not be brought to China.			w14081258_perova	0
+Accepted and revised in Figure1. All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11. The degradation of these isomers were stable within the former several days, which mentioned in the manuscript. So these former points conformed to first-order kinetic formula.	2	1	Actually, the soil samples were 0.25 mm sieved.			w14081258_perova	0
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar. Worse more, the soil can not be brought to China. Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential. But I did not do that. My visiting time is only one year. At the end, I had no time to complete and analysis the microbial community and biomass of the soil samples. But in the further study of the NP isomers in reclaimed water soil, this should be taken into account.	2	1	Your careful review of this manuscript is highly appreciated.			w14081258_perova	0
+Actually, the soil samples were 0.25 mm sieved. At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction. Therefore, 0.3mm-sieved soil was not used for the experiment(which has been deleted in the manuscript in L104). Then the soil was 0.25mm-sieved, which was suitable for the NP extraction. If the soil was 2 mm-sieved, the extraction of the NP was more inappropriate. So 0.25mm sieve was used in this study. Indeed, this size particle excluded an important part of the active soil where microorganisms live and organic carbon is accumulated. But for this study, the degradation was completed within 30 days in both of the two soils. Therefore, though some of the organic carbon was adsorbed on the surface of the particle, the amounts of the microorganisms are enough for the degradation of NP. However, the degradation rate could be affected by this. In the future study, this would be taken into accounted. What’s more, the microbial community could be taken into accounted as well. Accepted.	2	1	Migration and health risks of nonylphenol and bisphenol A in soil-winter wheat systems with long-term reclaimed water irrigation.			w14081258_perova	0
+Accepted and revised in L204. Figure 1 has been changed in the manuscript.	2	1	Ecotoxicology and Environmental Safety 158 (2018) 28–36.			w14081258_perova	0
+For example, there are some inconsistencies in having the first letter of some words in small and capital letters.			4) The abbreviated version of RCA has been used once before stating it with its extended version for the first time.			admsci5030125_boyarkin	0
+Also, you have to say “these types of incentives” as a plural version based on your used verb that is plural.			However, again, there is not sufficient amount of explanation/elaboration on how your version of the CIRA is connected to the original model.			admsci5030125_boyarkin	0
+3) You mentioned employing the “agency-theory” without mentioning anything about it.			4) The abbreviated version of RCA has been used once before stating it with its extended version for the first time.			admsci5030125_boyarkin	0
+You have Principle-agent problem on page 3 and Problem-Agent (Agent with capitalized “A”) on page 4.			Also, I believe that figure 2 needs some more explanation.			admsci5030125_boyarkin	0
+"page 4: you said “this type of incentives… (p.136)…” after “(p.136)"", you need a comma."			Then, it is followed by the model discussion, conclusion and future work.			admsci5030125_boyarkin	0
+However, again, there is not sufficient amount of explanation/elaboration on how your version of the CIRA is connected to the original model.			I have tried to accomodate your comments to the best of my ability.			admsci5030125_boyarkin	0
+From this point of view, this study can have a significant implication/application in the literature.			I have tried to accomodate your comments to the best of my ability.			admsci5030125_boyarkin	0
+This should change to “to manage the risk of human factors or to risk management of human factors”.			"page 4: you said “this type of incentives… (p.136)…” after “(p.136)"", you need a comma."			admsci5030125_boyarkin	0
+Also, you have to say “these types of incentives” as a plural version based on your used verb that is plural.			R4 You did a good job explaining your case study.			admsci5030125_makarova	0
+The CIRA does appear to be a useful and interesting tool for assessing risk, with particular regard to incentive structures.			From this point of view, this study can have a significant implication/application in the literature.			admsci5030125_makarova	0
+Based on what I see and get from reading your paper, the CIRA methodology is not the framework that you proposed.			You did cite a reference in this regard, but you may need to briefly explain it.			admsci5030125_makarova	0
+This should change to “to manage the risk of human factors or to risk management of human factors”.			On page 2 in the introduction the authors claim to apply CIRA to research questions such as finding the root cause of an incident of misconduct in peer review, as well as other potential risks inherent in the process.			admsci5030125_makarova	0
+I suggest that you review the whole paper again and edit it as needed.			You have some examples of what has been done in the area of peer review process and the risk analysis of such a process.			admsci5030125_makarova	0
+page 3, paragraph 5) while in other places, you said “peer-review”; hyphenated (e.g.			This does not look professional in scientific papers.			admsci5030125_makarova	0
+Thank you very much for your helpful commentary!			Second Round of Evaluation Round 2: and Author Response This is a significantly improved version of the manuscript.			admsci5030148_devetyaro	0
+This is unclear in the paper - a methodology chapter/discussion is needed.			Round 1: Author Response to Reviewer 3 I copied and pasted the reviewer commentary, below.			admsci5030148_devetyaro	0
+Second, the board-manager relationship is only a small part of the research on nonprofit boards, which in turn is the major focus of nonprofit governance research (Cornforth, 2012; Renz and Andersson, 2014).			To address these subtlies see: Turnbull (2002) A New Way to Govern Cornforth (2004) The Governance of Associations and Mutuals: A Paradox Perspective Ridley-Duff (2007) Communitarian Perspectives on Social Enterprise			admsci5030148_devetyaro	0
+In the social economy (associations, mutuals and cooperatives) the legal structures and accountabilities are different (and are captured well in the paper).			For example, the third sector itself can be seen as diverse with philanthropic, mutual, cooperative and association sub-sectors.			admsci5030148_devetyaro	0
+- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.			admsci5030148_devetyaro	0
+Just as the wealthy entrepreneur seeks to control the private enterprise through shareholdings, so the wealthy philanthropist seeks to control the non-profit enterprise through trust law.			Round 1: Author Response to Reviewer 4 I copied and pasted the reviewer commentary, below.			admsci5030148_devetyaro	0
+Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			These issues, and the second one in particular, points towards unclear research methodology.			admsci5030148_devetyaro	0
+A sectoral comparison of wage levels and wage inequality in human services industries.			These apply in both cases because funding is sought from wealthy providers with the power to frame laws that require their investees to use the money for the purposes stimpulated by them.			admsci5030148_devetyaro	0
+The comments at the start of the literature review might apply to foundations and charities, but are less true of association and mutuals (let alone cooperatives).			First, the principal-agent approach is only one of theoretical perspectives used to analyze nonprofit governance (Cornforth, 2004; Renz and Andersson, 2014).			admsci5030148_devetyaro	0
+- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			The text itself is now almost finished, I only have a few minor comments: -			admsci5030148_devetyaro	0
+Nonprofit governance research: Limitations of the focus on boards and suggestions for new directions.			Your choices may be OK in given circumstances, but how do you argue for these choices here?			admsci5030148_devetyaro	0
+This is unclear in the paper - a methodology chapter/discussion is needed.			- the case description and analysis is based only on media accounts - no actual first hand information to inform the analysis.			admsci5030148_devetyaro	0
+R3 These issues, and the second one in particular, points towards unclear research methodology.			- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			admsci5030148_devetyaro	0
+The writing is good, there is a sound development of the theory, and some well drawn conclusions.			Round 1: Author Response to Reviewer 4 I copied and pasted the reviewer commentary, below.			admsci5030148_devetyaro	0
+Minor issues (details really): - Wrong reference number on page 4, line 14: should be 15.			In the abstract, the author also states that the over-reliance on the principal-agent model introduces two challenges.			admsci5030148_devetyaro	0
+Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			COMMENT #4: The comments added in the beginning of Chapter 3 on methodology are important and improves the text a lot.			admsci5030148_devetyaro	0
+R3 These issues, and the second one in particular, points towards unclear research methodology.			The text itself is now almost finished, I only have a few minor comments: - The literature part is improved by changing the headline, putting it in the right setting.			admsci5030148_devetyaro	0
+Furthermore, given that the world of third sector organizations is very rich and heterogeneous, one should be careful by discussing third sector organizations in general, especially since the case study seems to focus on co-operative enterprises.			However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles?			admsci5030148_devetyaro	0
+Your comments have made this paper significantly better.			The author openly acknowledges that the basis for a complete picture is lacking.			admsci5030148_devetyaro	0
+R12 The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.			The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation.			admsci5030148_devetyaro	0
+Nonprofit governance research: Limitations of the focus on boards and suggestions for new directions.			A sectoral comparison of wage levels and wage inequality in human services industries.			admsci5030148_devetyaro	0
+I do not agree with the author for several reasons.			Secondly, I just can't agree that third sector governance is derived from for-profit principles.			admsci5030148_devetyaro	0
+The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation.			Round 1: Author Response to Reviewer 1 I copied and pasted the reviewer commentary, below.			admsci5030148_devetyaro	0
+I do not agree with the author for several reasons.			Your comments have made this paper significantly better.			admsci5030148_devetyaro	0
+Is there a way to interview members who participated in action (or access sources written by them)?			However, there are subtleties in the literature that need considering, and also the methodology (which is extremely weak).			admsci5030148_devetyaro	0
+Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal R9 structures and accountability mechanisms reflect this.			Nonprofit and Voluntary Sector Quarterly 40(4): 608-633.			admsci5030148_devetyaro	0
+It is good to see the author is open for suggestions from reviewers.			Cornforth (2004) has already analyzed the governance of co-operatives from multiple theoretical R5 perspectives.			admsci5030148_devetyaro	0
+Nonprofit and Voluntary Sector Quarterly 41(3): 431-451.			In addition, a more extensive literature review and comparison with previous principal-agent literature and stakeholder governance models (especially Van Puyvelde et al., 2012 and Coule, 2015) may give the manuscript a more solid theoretical foundation.			admsci5030148_devetyaro	0
+I still have doubts about the basis for conclusion and generalization of findings.			First, the principal-agent approach is only one of theoretical perspectives used to analyze nonprofit governance (Cornforth, 2004; Renz and Andersson, 2014).			admsci5030148_devetyaro	0
+Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.			THEORISATION Lastly, the discussion and conclusions need to stick to contributions to theory / research practice and refrain from commenting on wider issues without adequate justification.			admsci5030148_devetyaro	0
+I do not agree with the author for several reasons.			Is there a way to interview members who participated in action (or access sources written by them)?			admsci5030148_devetyaro	0
+"COMMENT #6: Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"			This paper communicated a nice research idea, but I find it too superficial in its approach.			admsci5030148_devetyaro	0
+The conceptual framework is not clearly presented and I do not see what are the contributions except some free statements at the end not really related to the analysis.			Given the aforementioned comments, I would advise the author to revise the paper by focusing on the governance of nonprofit associations / co-operatives.			admsci5030148_devetyaro	0
+Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal R9 structures and accountability mechanisms reflect this.			However, the credibility of the R11 paper and findings would be enormously enhanced is some primary interview data could be collected and presented.			admsci5030148_devetyaro	0
+I would not have thought of this as a major issue unless Evers and Laville (2004) The third sector in Europe, Edward Elgar, spesifically discuss whether cooperations are defined within or outside the third sector.			The text itself is now almost finished, I only have a few minor comments: - The literature part is improved by changing the headline, putting it in the right setting.			admsci5030148_devetyaro	0
+Certainly, some description of the process by which the case study was developed (sources used, data collection techniquies, analysis techniques) are needed to be publishable.			The governance of cooperatives and mutual associations: A paradox perspective.			admsci5030148_devetyaro	0
+(2012) and the idea that is necessary to capture the broader stakeholder environment in a democratic way is also not new (Coule, 2015).			This is unclear in the paper - a methodology chapter/discussion is needed.			admsci5030148_devetyaro	0
+Nonprofit Management & Leadership 12(4): 429 -450.			Minor issues (details really): - Wrong reference number on page 4, line 14: should be 15.			admsci5030148_makarova	0
+It is good to see the author is open for suggestions from reviewers.			However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles?			admsci5030148_makarova	0
+The board as a monitor of organizational activity: the applicability of agency theory to nonprofit boards.			Furthermore, attempts have already been made to broaden the principal-agent approach in nonprofit organizations (Ben-Ner et al., 2012; Van Puyvelde et al., 2012, Coule, 2015), leading me to the question what this manuscript actually adds to the literature.			admsci5030148_makarova	0
+The use of media sources, while not ideal, might be adequate if the author(s) can show systematic and comprehensive collection of them.			The author openly acknowledges that the basis for a complete picture is lacking.			admsci5030148_makarova	0
+Minor issues (details really): - Wrong reference number on page 4, line 14: should be 15.			The author openly acknowledges that the basis for a complete picture is lacking.			admsci5030148_makarova	0
+- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			R12 The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.			admsci5030148_makarova	0
+There is, however, some common practices, but not for the reasons described.			Associations, mutuals and cooperatives, however, are not dependent on the patronage of the rich, but the patronage of the many, and their mass member legal R9 structures and accountability mechanisms reflect this.			admsci5030148_makarova	0
+I still have doubts about the basis for conclusion and generalization of findings.			I believe that this then addresses the issues that fall in line with your critiques about the theoretical foundation.			admsci5030148_makarova	0
+It is very similar to the nonprofit governance model of Van Puyvelde et al.			Second Round of Evaluation Round 2: and Author Response This is a significantly improved version of the manuscript.			admsci5030148_makarova	0
+"COMMENT #6: Low on page 16: ""The ownership becomes absentee, ..."" This is in the case of co-operative?"			LITERATURE R10 The characterisation of third sector governance is too crude.			admsci5030148_makarova	0
+- USA context concerning cooperatives (upper half page 10) should be highlighted even more for readers to be able to compare to their own context (for instance in Europe)			Stick to commenting on the usefulness and value of ODPs, and draw out any theoretical contributions regarding the design principles themselves, or their value as a theoretical perspective for governance research.			admsci5030148_makarova	0
+A sectoral comparison of wage levels and wage inequality in human services industries.			In both cases, Principle-Agent assumptions apply, but not because both are based on for-profit assumptions.			admsci5030148_makarova	0
+Thank you very much for your helpful commentary!			Round 1: and Author Response The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			admsci5030148_makarova	0
+Furthermore, given that the world of third sector organizations is very rich and heterogeneous, one should be careful by discussing third sector organizations in general, especially since the case study seems to focus on co-operative enterprises.			The disadvantages of a too narrow principal-agent approach have been acknowledged in previous studies (Miller, 2002).			admsci5030148_makarova	0
+Some well grounded comments on their use in other governance research are merited right at the end.			I also have problems with the depth of the discussion, and I feel the conclusion does not match the promises of the abstract.			admsci5030148_makarova	0
+I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).			Round 1: Author Response to Reviewer 1 I copied and pasted the reviewer commentary, below.			admsci5030148_makarova	0
+Second, I note in the paper that I am not making the claim that the co-operative model is part of the third sector.			However, a solid foundation for this argument is missing.			admsci5030148_makarova	0
+The disadvantages of a too narrow principal-agent approach have been acknowledged in previous studies (Miller, 2002).			I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).			admsci5030148_makarova	0
+Stick to commenting on the usefulness and value of ODPs, and draw out any theoretical contributions regarding the design principles themselves, or their value as a theoretical perspective for governance research.			Round 1: Author Response to Reviewer 4 I copied and pasted the reviewer commentary, below.			admsci5030148_makarova	0
+- The comments added in the beginning of Chapter 3 on methodology are important and improves the text a lot.			- Figure 1 text overwrite part of the figure, and there should be comments on how this is different from a general structure for a private sector company (if you look at stakeholders as shareholders).			admsci5030148_makarova	0
+Voluntas 22:566–586 R6 Round 1: Author Response to Reviewer 2 I copied and pasted the reviewer commentary, below.			Given the inclusion of Coule's paper, I would have already expected more sensitivity to divisions unitary and pluralist theories of governance but by reviting this (and reading Turnbull's Cornforth's and Ridley-Duff's precursor to Coule's work), this should be adequately addressed.			admsci5030148_makarova	0
+First, the principal-agent approach is only one of theoretical perspectives used to analyze nonprofit governance (Cornforth, 2004; Renz and Andersson, 2014).			R7 I am also not convinced by the definition of co-operatives mentioned in the paper.			admsci5030148_makarova	0
+Given the case study, I would focus on the governance of co-operatives (Cornforth, 2004).			As such, applying the idea of the Ostrom design principles to nonprofit governance and illustrating this with a case study may more clearly show the contribution of the manuscript to the current literature.			admsci5030148_makarova	0
+This is unclear in the paper - a methodology chapter/discussion is needed.			- The cooperative firm removes adversarial aspects of market contracting (page 11) - what adversarial aspects?			admsci5030148_makarova	0
+This is unclear in the paper - a methodology chapter/discussion is needed.			The abstract of the manuscript mentions that dominant modes of third sector organizational governance adhere to a narrow principal-agent orientation.			admsci5030148_makarova	0
+Given the case study, I would focus on the governance of co-operatives (Cornforth, 2004).			R8 Note that the paper no longer claims this to be a case study, and instead positions itself as a discussion paper, performing a secondary archival analysis of media accounts.			admsci5030148_makarova	0
+), Nonprofit Governance: Innovative Perspectives and Approaches.			I do not understand the purpose of the paper.			admsci5030148_makarova	0
+Voluntas 22:566–586 R6 Round 1: Author Response to Reviewer 2 I copied and pasted the reviewer commentary, below.			R12 The author is mixing up several concepts and ignore the abundant literature on governance in nonprofits not dealing with the agency theory.			admsci5030148_makarova	0
+Stick to commenting on the usefulness and value of ODPs, and draw out any theoretical contributions regarding the design principles themselves, or their value as a theoretical perspective for governance research.			Nonprofit and Voluntary Sector Quarterly 41(3): 431-451.			admsci5030148_makarova	0
+However, I cannot see how chapter 2.2. is a sub-theme under the Ostrom Principles?			I think it needs linking back to the findings more clearly and to be elaborated and slightly more length (cut some literature if needed to stay within the word length limits).			admsci5030148_makarova	0
+The basic premise of this article is good: the application of Ostrom's design principles as a way of analysing / evaluating governance in third sector organisations is quite innovative, particularly combined with a discussion of third sector / cooperative governance principles.			Cornforth (2004) has already analyzed the governance of co-operatives from multiple theoretical perspectives.			admsci5030148_makarova	0
+The authors conclude that they found ‘consistent combinations of the types of national policies, local clusters, and cluster management’.			Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.			admsci5040213_boyarkin	0
+The relevance of the information presented here for the following discussion and conclusion remain far from clear.			• Page 10, line 19 – ‘several cluster policies’: I would address this family of related programmes as one policy rather than many.			admsci5040213_boyarkin	0
+Given the conceptual framework outlined in figure 1, one might also wonder why national policymakers, i.e.			(…) A conceptual and/or theoretical perspective is needed to shed light at the differences found.			admsci5040213_boyarkin	0
+in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up.			As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing.			admsci5040213_boyarkin	0
+‘Public research-led cluster’ might be more to the point, as it appears difficult to judge if these are more or less ‘intellectual’ than other forms of clusters.			When outlining their research methodology, the authors should state precisely how many interviews they conducted.			admsci5040213_boyarkin	0
+Section 4: The presentation of the six case studies is very systematic and clear.			Broad Comments The paper fills an important research deficit by focusing on the interdependence between national cluster policy programmes and their implementation by cluster management on the ground.			admsci5040213_boyarkin	0
+Since Japan, France and Germany are all classified as co-ordinated market economies in the original VoC concept, more nuanced differentiations of capitalisms – or other institutional frameworks – might be recommended for the paper.			Second Round of Evaluation Round 2: The paper now looks but			admsci5040213_boyarkin	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			The performance of clusters is also affected by many exogenous influences not captured in the simple model.			admsci5040213_boyarkin	0
+o P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably R15 taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?			Broad Comments • However, the conceptual framework appears simplistic, technocratic and static, and it is not sufficiently rooted in literature as evident from the scarcity of references in section 2.2.			admsci5040213_boyarkin	0
+• Section 2 contains a number of “expectations”, which could be flagged out and numbered as hypotheses.			The performance of clusters is also affected by many exogenous influences not captured in the simple model.			admsci5040213_boyarkin	0
+As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing.			; BOSCHMA, R.; COOKE, P., 2011: Constructing regional advantage: Platform policies based on related variety and differentiated knowledge bases.			admsci5040213_boyarkin	0
+Also sound profiling of the different cluster programs and initiatives.			R2 Round 1: Author Response to Reviewer 1 We are very grateful for your valuable comments and suggestions, which help improve our manuscript significantly.			admsci5040213_boyarkin	0
+KOSCHATZKY, K., 2000: A river is a river – Cross-border networking between Alsace and Baden.			We provided minor corrections to our manuscript based on the following comments.			admsci5040213_boyarkin	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			R14 Round 2: I wish to congratulate the authors for their careful and thoughtful revision.			admsci5040213_boyarkin	0
+We tried to consider the comments and suggestions as far as possible in revising our manuscript.			In sum, I recommend that the paper be reconsidered after a major revision.			admsci5040213_boyarkin	0
+• The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies.			There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti).			admsci5040213_boyarkin	0
+As Kiese (2009) showed for three European Countries including Germany and France, these differences leave an imprint on the design and implementation of national cluster policies.			The varieties of capitalism (VoC) concept developed by HALL/SOSKICE (2011) may be a suitable concept here, as it has been applied to cluster policies in the U.S. and Germany by STERNBERG ET AL.			admsci5040213_boyarkin	0
+When outlining their research methodology, the authors should state precisely how many interviews they conducted.			1992: National Systems of Innovation: Towards a Theory of Innovation and Interactive Learning.			admsci5040213_boyarkin	0
+Round 2: Author Response to Reviewer 2 I wish to congratulate the authors for their careful and thoughtful revision.			Since this may be managed by the editorial office, I do not see the need for another review.			admsci5040213_boyarkin	0
+• Furthermore, ‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance.			Round 1: Author Response to Reviewer 2 We are very grateful for your detailed, insightful comments and suggestions, which help improve our manuscript significantly.			admsci5040213_boyarkin	0
+R10 · Section 4: The presentation of the six case studies is very systematic and clear.			• Page 15, line 27: Repetition – the IZB incubator was already mentioned on line 9.			admsci5040213_boyarkin	0
+As a consequence, I can now recommend the paper for publication with minor corrections, including some language editing.			If ‘few’ means ‘no’, then please say so straightaway.			admsci5040213_boyarkin	0
+R14 Round 2: I wish to congratulate the authors for their careful and thoughtful revision.			; Soete, L. (eds): Technical Change and Economic Theory.			admsci5040213_boyarkin	0
+BRETTE, O.; CHAPPOZ, Y., 2007: The French competitiveness clusters: Toward a new public policy for innovation and research?			Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique.			admsci5040213_boyarkin	0
+For the German cases, the authors may have underestimated public agency, since they did not investigate the role of state governments (Bavaria, Baden-Württemberg) within the country’s system of multilevel governance, which is key to understand cluster policies in Germany (cf.			A substantial body of literature has grown on the back of seminal contributions by Edquist (1997), Freeman (1987, 1988, 1995) with reference to Japan, Lundvall (1992), and Nelson (1993).			admsci5040213_boyarkin	0
+In: European Review of Industrial Economics and Policy (5).			• Finally, the paper is static as it does not account for cluster evolution and (policy) learning over time.			admsci5040213_boyarkin	0
+P. 14: On section 4.5 in the first draft, I commented that “Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that Koschatzky (2000) had identified long before.			R14 Round 2: I wish to congratulate the authors for their careful and thoughtful revision.			admsci5040213_boyarkin	0
+Such limitations do not necessarily disqualify the paper, but should be discussed reflexively in the paper’s conclusions.			According to the definition on the top of page 4, the term denotes a R9 cluster led by a public research organisation, which is rather specific and I doubt it applies to the six case studies outlined in the paper.			admsci5040213_boyarkin	0
+In the following, we will reply to each of your comments, using blue letters for our responses (underlines in the comments by the authors).			• Page 15, lines 22-23: ‘Large firms are on the whole not active in clusters’ – As a general statement, this is certainly not true.			admsci5040213_boyarkin	0
+First Round of Evaluation Round 1: Interesting idea to look at the impact of cluster policy/program design on the cluster (initiative management).			o P. 22: “differences might be attributed to those in innovation systems as basic conditions of clusters.” This is central, so the discussion should be summarised in the conclusions, preferably R15 taking up the three hypotheses: To what extent and how can differences in cluster policies and cluster management be linked to differences in national systems of innovation?			admsci5040213_boyarkin	0
+If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences.			We tried to consider the comments and suggestions as far as possible in revising our manuscript.			admsci5040213_boyarkin	0
+We tried to consider the comments and suggestions as far as possible in revising our manuscript.			• Section 4.5: Unlike the regional cluster initiatives in Alsace and Baden, the trinational BioValley initiative has hardly had any impact in terms of intensifying cross-border interaction, confirming problems that KOSCHATZKY (2000) had identified long before.			admsci5040213_boyarkin	0
+In: European Review of Industrial Economics and Policy (5).			When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!			admsci5040213_boyarkin	0
+In: European Review of Industrial Economics and Policy (5).			KOSCHATZKY, K., 2000: A river is a river – Cross-border networking between Alsace and Baden.			admsci5040213_boyarkin	0
+; Soete, L. (eds): Technical Change and Economic Theory.			The conceptual framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included.			admsci5040213_boyarkin	0
+KIESE, M., 2009: National styles of cluster promotion: Cluster policies between variety and convergence.			Even so, the term ‘intellectual’ does not look fully appropriate here, which may be due to its translation from a Japanese original.			admsci5040213_boyarkin	0
+In: Journal of Economic Issues, 41(2): 391-398.			This appears hardly surprising given the co-evolution of policies and cluster management within a common institutional environment.			admsci5040213_boyarkin	0
+Would suggest to use these case studies as a source of ideas on how policies and cluster initiative management might relate to each other; that would make this a more interesting paper.			In the following, we will reply to each of your comments, using blue letters for our responses.			admsci5040213_boyarkin	0
+At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.			• Finally, the paper is static as it does not account for cluster evolution and (policy) learning over time.			admsci5040213_boyarkin	0
+KIESE, M., 2009: National styles of cluster promotion: Cluster policies between variety and convergence.			The performance of clusters is also affected by many exogenous influences not captured in the simple model.			admsci5040213_boyarkin	0
+Round 1: Referee Report for Administrative Sciences Management of cluster policies for innovation: Comparative case studies of Japanese, German, and French biotechnology clusters Manuscript no.			In: European Review of Industrial Economics and Policy (5).			admsci5040213_boyarkin	0
+If the underlying institutional differences would have been revealed as suggested, this section would be even more valuable for understanding the differences.			In: European Review of Industrial Economics and Policy (5).			admsci5040213_boyarkin	0
+The authors conclude that they found ‘consistent combinations of the types of national policies, local clusters, and cluster management’.			"There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."			admsci5040213_boyarkin	0
+KIESE, M., 2009: National styles of cluster promotion: Cluster policies between variety and convergence.			Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.			admsci5040213_boyarkin	0
+• Finally, the paper is static as it does not account for cluster evolution and (policy) learning over time.			The conceptual framework (figure 1) is very similar to the 'cluster initiative performance model' in the Cluster Initiative Greenbook (Solvell et al., 2003), adding the hypothesis that there might be an interaction between policy context and management and being somewhat more narrow on the elements included.			admsci5040213_devetyaro	0
+Round 2: Author Response to Reviewer 2 I wish to congratulate the authors for their careful and thoughtful revision.			It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.			admsci5040213_devetyaro	0
+How can these be ‘representative’ for whatever population of biotech clusters in these countries?			P. 16: Sources should be stated underneath table 2.			admsci5040213_devetyaro	0
+• It is simplistic and technocratic because it views cluster performance as an outcome of initial conditions (i.e., the state of cluster development), cluster management and national polices.			At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.			admsci5040213_devetyaro	0
+This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation.			regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.			admsci5040213_devetyaro	0
+Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			In the following, we will reply to each of your comments, using blue letters for our responses.			admsci5040213_devetyaro	0
+There are also existing reviews of cluster policies in Europe that should be considered (Clusters are Individuals, 2012, BMWi; VDI/VDE; Dasti).			For instance, what does it mean if a regional cluster organisation is pre-dating the national policy programme it is supposed to implement, as evident from some of the cases?			admsci5040213_devetyaro	0
+in terms of the role of the state versus private initiative, governance traditions and philosophies, centralist vs. federal set-up.			It seems that the most prominent or successful cases have been selected, at least for Germany and France.			admsci5040213_devetyaro	0
+Section 4: The presentation of the six case studies is very systematic and clear.			Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.			admsci5040213_devetyaro	0
+• Section 5 does not include any reference and is purely descriptive and sometimes even superficial, lacking any comparison, explanation or assessment.			R7 LONGHI, C.; ROCHHIA, S., 2013: Cluster policy for innovation and competitiveness: Lessons from the French experience.			admsci5040213_devetyaro	0
+The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies.			In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.			admsci5040213_devetyaro	0
+• Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.			The performance of clusters is also affected by many exogenous influences not captured in the simple model.			admsci5040213_devetyaro	0
+Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique.			Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique.			admsci5040213_devetyaro	0
+• Page 12, lines 11-12: Table 2 provides a descriptive summary, but no ‘explanation’ – see major comments.			Section 4: The presentation of the six case studies is very systematic and clear.			admsci5040213_devetyaro	0
+Is there comparable evidence on policy learning between Germany and France?			Compared to the Japanese cases, the French and German cases are clearly more bottom-up, but they still display clear – if not dominant – elements of top-down governance and public agency.			admsci5040213_devetyaro	0
+Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			• Methodology: The choice of clusters is generally well explained on pp.			admsci5040213_devetyaro	0
+), 1993: National Systems of Innovation: A Comparative Study.			For the German cases, the authors may have underestimated public agency, since they did not investigate the role of state governments (Bavaria, Baden-Württemberg) within the country’s system of multilevel governance, which is key to understand cluster policies in Germany (cf.			admsci5040213_devetyaro	0
+KIESE, M., 2013: Regional cluster policies in Germany – A multi-level governance perspective on policy learning.			Round 1: Author Response to Reviewer 2 We are very grateful for your detailed, insightful comments and suggestions, which help improve our manuscript significantly.			admsci5040213_devetyaro	0
+(…) The varieties of capitalism (VoC) concept developed by HALL/SOSKICE (2011) may be a suitable concept here, as it has been applied to cluster policies in the U.S. and Germany by STERNBERG et al.			Indeed, many INTERREG projects fail to overcome the differences in national languages, legislations and cultures, even though the regions are neighbours.” In response, the authors now cite this paper in footnote 14 in Section 3.2. to support the view that BioValley has hardly had any impact in terms of intensifying cross-border interaction.			admsci5040213_devetyaro	0
+The authors might find the following major and minor comments helpful when revising their paper.			I would therefore suggest to drop this footnote altogether.			admsci5040213_devetyaro	0
+When elaborating on the choice of nations, one might wonder why the United States have not been included – possibly because there is no comparable national cluster policy?!			This should be harmonised, or preferably conversions given in brackets or footnotes for the reader’s convenience.			admsci5040213_devetyaro	0
+Furthermore, the state government of R5 Bavaria may be a regional government, but it is certainly not a local authority (like the city of Munich; page 15, line 29) in Germany’s system of multilevel governance.			However, it remains rather descriptive and does not systematically attempt to explain the differences found between the three countries.			admsci5040213_devetyaro	0
+• Furthermore, ‘initial conditions’ fail to capture the complexity of clusters if they are reduced to a dichotomy of private vs. public sector dominance.			regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.			admsci5040213_devetyaro	0
+; Soete, L. (eds): Technical Change and Economic Theory.			It develops a conceptual frame for the comparative analysis of cluster policies and applies it to six case studies in three countries.			admsci5040213_devetyaro	0
+How can these be ‘representative’ for whatever population of biotech clusters in these countries?			Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_devetyaro	0
+(…) The varieties of capitalism (VoC) concept developed by HALL/SOSKICE (2011) may be a suitable concept here, as it has been applied to cluster policies in the U.S. and Germany by STERNBERG et al.			Freeman, C., 1988: Japan: A new National System of Innovation?			admsci5040213_devetyaro	0
+The varieties of capitalism (VoC) concept developed by HALL/SOSKICE (2011) may be a suitable concept here, as it has been applied to cluster policies in the U.S. and Germany by STERNBERG ET AL. (2010).			Furthermore, the state government of R5 Bavaria may be a regional government, but it is certainly not a local authority (like the city of Munich; page 15, line 29) in Germany’s system of multilevel governance.			admsci5040213_devetyaro	0
+STERNBERG, R.; KIESE, M.; STOCKINGER, D., 2010: Cluster policies in the US and Germany: Varieties of capitalism perspective on two high-tech states.			; SOSKICE, D., 2001: An introduction to varieties of capitalism.			admsci5040213_devetyaro	0
+It certainly helps preventing, partly prevents, or reduces the risk of these problems occurring.			The authors focus on what they call ‘intellectual clusters’, a term that appears 21 times throughout the paper although I cannot recall having ever seen it in a decade or so of studying clusters and cluster policies.			admsci5040213_devetyaro	0
+), 1993: National Systems of Innovation: A Comparative Study.			How can these be ‘representative’ for whatever population of biotech clusters in these countries?			admsci5040213_devetyaro	0
+Freeman, C., 1995: The National System of Innovation in Historical Perspective.			• The paper contributes to comparative cluster policy research and thus helps overcoming the prevailing focus on individual case studies.			admsci5040213_devetyaro	0
+"There is also a growing literature on cluster initiative management in Europe (see the EU's ""Excellence Initiative' with the benchmarking of cluster initiatives) that should be considered."			), 1993: National Systems of Innovation: A Comparative Study.			admsci5040213_devetyaro	0
+How can these be ‘representative’ for whatever population of biotech clusters in these countries?			• Page 14, line 21: How can this be a cluster if there are ‘only a few biotech ventures’?			admsci5040213_devetyaro	0
+the interface between national cluster policies and cluster management on the ground, and thus makes an original contribution to our understanding of cluster policies and their implementation.			In my humble opinion, this is the weakest part of the paper that might well be omitted and replaced by a suitable conceptual frame for international comparison, as outlined above.			admsci5040213_devetyaro	0
+Timing and content suggest that French national policies may well have been inspired by the German BioRegio contest.			Critical issues such as the cluster management organisation’s budget and finance have not been addressed.			admsci5040213_devetyaro	0
+At the least, it would appear sound to qualify these six cases as science-based clusters, but then this would apply to the biotech industry and all its clusters in general.			KIESE, M., 2009: National styles of cluster promotion: Cluster policies between variety and convergence.			admsci5040213_devetyaro	0
+Although ‘top-down’ and ‘bottom-up’ have been replaced by public-driven and ‘private-driven’, the dichotomist nature remains as the focus of my critique.			It develops a conceptual frame for the comparative analysis of cluster policies and applies it to six case studies in three countries.			admsci5040213_devetyaro	0
+Since Japan, France and Germany are all classified as co-ordinated market economies in the original VoC concept, more nuanced differentiations of capitalisms – or other institutional frameworks – might be recommended for the paper.			The authors should include a discussion on how these R4 countries differ e.g.			admsci5040213_devetyaro	0
+regarding their specialisation within biotechnology, their maturity, size (number of firms and employee) and firm size structure.			This section now uses the term ‘innovation system’ at the national scale, but there is no discussion of or reference to the concept of national systems of innovation.			admsci5040213_devetyaro	0
+In: Journal of Economic Issues, 41(2): 391-398.			In: Environment and Planning C, 28(6): 1063-1082.			admsci5040213_devetyaro	0
+Compared to the Japanese cases, the French and German cases are clearly more bottom-up, but they still display clear – if not dominant – elements of top-down governance and public agency.			; BOSCHMA, R.; COOKE, P., 2011: Constructing regional advantage: Platform policies based on related variety and differentiated knowledge bases.			admsci5040213_devetyaro	0
+At present, the final section is merely a summary with the exception of the very last sentence briefly sketching the need for including evaluation in comparative cluster policy research.			BRETTE, O.; CHAPPOZ, Y., 2007: The French competitiveness clusters: Toward a new public policy for innovation and research?			admsci5040213_devetyaro	0
+Would suggest to use these case studies as a source of ideas on how policies and cluster initiative management might relate to each other; that would make this a more interesting paper.			I would still like to see a qualification stating that R16 private and public initiative often interacts in cluster development, and that the degree of private and public sector involvement usually differs from case to case.			admsci5040213_devetyaro	0
+It seems that the most prominent or successful cases have been selected, at least for Germany and France.			http://revel.unice.fr/eriep/index.html?id=3495, accessed June 13, 2015.			admsci5040213_devetyaro	0
+Methodology (a) Though the paper is exploratory, I am not convinced that it aligns with the “grounded theory” tradition.			That being said, I believe there is an opportunity to improve the paper by developing the conceptual foundations more thoroughly and my being more transparent and clear in the methodology section.			admsci6020005_makarova	0
+Though such studies are rare, good interpretive and quantitative examples can be found in the literature.			While Jungian approaches have been used by marketing and consumer researchers in the past (e.g., Hirschman), the reasons for this choice need to be justified in the context of other alternatives.			admsci6020005_makarova	0
+At a minimum, this exploratory paper should set a more specific and ambitious plan for systematic future research on the topic.			After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity.			admsci6020005_makarova	0
+Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes.			This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.			admsci6020005_makarova	0
+"For example the fact that some ""heroes"" can move across categories - e.g., Tiger Woods for Buick."			The recommendations were very constructive and we hope that we have addressed all.			admsci6020005_makarova	0
+These ideas in developed with (some of) the relevant literature and are demonstrated using a simple two-firm case study.			Time of message delivery and personal relevance to the targeted audience may be ( I believe) of more importance than the creative.			admsci6020005_makarova	0
+Linda M. Scott and Rajeev Batra, Mahwah, NJ: Lawrence Erlbaum Associates, 223-45.			Personally, I agree with you – brand personality is a key facet of brand identity.			admsci6020005_makarova	0
+After all, the research focuses on the capacity of an ad to convey archetypal and personality-based aspects of brand identity.			We are delighted that our reviewer sees the link (as the authors do) between Aaker and Jung’s images of Freedom, Social, Order and Ego.			admsci6020005_makarova	0
+Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).			Please don’t expect them to consult original sources by Jung!			admsci6020005_makarova	0
+Are there any other limitations or criticisms the reader should be informed of?			Why do you not report reliability measures for the items and scale?			admsci6020005_makarova	0
+The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.			I hope my suggestions are given full consideration, as I believe that they will assist them in achieving this important objective.			admsci6020005_makarova	0
+The authors have selected Jung’s archetypes as the theoretical foundation for the paper.			However, if you included a storyboard (set of screenshots) for each ad, that would help the reader (for examples, see: Mulvey and Medina 2003; Scott 1994).			admsci6020005_makarova	0
+Reviewing the famous Marlbro ads or thinking about the attention David Beckham's appearance in ads for Adidas receives can be understood to represent viewer's interpretation and unconscious assignment of famous archetypes.			Imagine a conversation with these authors: what do you contribute to the ongoing conversation?			admsci6020005_makarova	0
+Page 4, line 124 states: “European students did not know these two hotel chains and therefore were not pre-conditioned in any way.			The results would have been more compelling if the study measured the impact of ad campaigns (sets of related ads) in delivering a consistent (reliable) impression.			admsci6020005_makarova	0
+We are delighted that our reviewer sees the link (as the authors do) between Aaker and Jung’s images of Freedom, Social, Order and Ego.			Author Response Author Response File: Author Response.docx			admsci6020005_makarova	0
+Whereas the BT commercials features beautiful pictures that are sequence-wise interchangeable, the BT commercial unfolds and has it climax at the end with the wolves warming the traveler.			Most Western executives stay at well-known global hotel chains (e.g., Accor, Hilton) because they have a corporate contract.			admsci6020005_makarova	0
+"Padgett, Dan and Douglas Allen (1997), ""Communicating Experiences: A Narrative Approach to Creating Service Brand Image,"" Journal of Advertising, 26 (4), 49-62."			The recommendations were very constructive and we hope that we have addressed all.			admsci6020005_perova	0
+Time of message delivery and personal relevance to the targeted audience may be ( I believe) of more importance than the creative.			Linda M. Scott and Rajeev Batra, Mahwah, NJ: Lawrence Erlbaum Associates, 223-45.			admsci6020005_perova	0
+At a minimum, this exploratory paper should set a more specific and ambitious plan for systematic future research on the topic.			As mentioned at the onset of my review, I fully support the authors’ aim to revitalize and blend archetypal and personality-based approaches to brand identity.			admsci6020005_perova	0
+I think that the relevance of the research to practitioners as well as academics needs to include some current findings about ads, their placement and effectiveness.			Author Response Author Response File: Author Response.docx			admsci6020005_perova	0
+"--- (1989), ""Literary Criticism and Consumer Research: Overview and Illustrative Analysis,"" Journal of Consumer Research, 16 (3), 322-34."			Personally, I agree with you – brand personality is a key facet of brand identity.			admsci6020005_perova	0
+(h) Factor analysis: Have you reported principal components, or a rotated solution?			"Padgett, Dan and Michael S. Mulvey (2007), ""Differentiation Via Technology: Strategic Positioning of Services Following the Introduction of Disruptive Technology,"" Journal of Retailing, 83 (4), 375-91."			admsci6020005_perova	0
+The authors have selected Jung’s archetypes as the theoretical foundation for the paper.			"Padgett, Dan and Michael S. Mulvey (2007), ""Differentiation Via Technology: Strategic Positioning of Services Following the Introduction of Disruptive Technology,"" Journal of Retailing, 83 (4), 375-91."			admsci6020005_perova	0
+The recommendations were very constructive and we hope that we have addressed all.			The general and growing aversion to ads - ad blocker or the increased use of native advertising - sponsored branded content - that appears in desired media channels is a major issue for ad agencies.			admsci6020005_perova	0
+Author Response Author Response File: Author Response.docx			Aaker’s approach is nested within these – it specifies a way brands can establish relevance in the eyes of consumers (via establishing a human identity or character).			admsci6020005_perova	0
+I think there are some ways in which you may tighten up the description of the experimental design.The comparisons between Jung and Aaker are always interesting - although the small sample size may be troubling to some.			Though such studies are rare, good interpretive and quantitative examples can be found in the literature.			admsci6020005_perova	0
+Imagine a conversation with these authors: what do you contribute to the ongoing conversation?			(b) Sample size: A critical view would state that the present research relies on a very small sample of advertisements (n=2) to make its claims.			admsci6020005_perova	0
+The claim is very absolute – I wonder if any of them ever travelled internationally – presumably EMBAs would be more worldly or cosmopolitan than this…			"Padgett, Dan and Michael S Mulvey (2009), ""Experiential Positioning: Strategic Differentiation of Customer-Brand Relationships,"" Innovative Marketing, 5 (3), 87-95."			admsci6020005_perova	0
+"Padgett, Dan and Douglas Allen (1997), ""Communicating Experiences: A Narrative Approach to Creating Service Brand Image,"" Journal of Advertising, 26 (4), 49-62."			"Mulvey, Michael S. and Carmen Medina (2003), ""Invoking the Rhetorical Power of Character to Create Identifications,"" in Persuasive Imagery: A Consumer Response Perspective, ed."			admsci6020005_perova	0
+The recommendations were very constructive and we hope that we have addressed all.			Author Response Author Response File: Author Response.docx			admsci6020005_perova	0
+This paper’s potential impact will be enhanced if it can find a way to align itself with this growing branch of advertising research.			One could make the argument that using archetypes in advertising has greater affinities to mythology, literature and communications.			admsci6020005_perova	0
+These ideas in developed with (some of) the relevant literature and are demonstrated using a simple two-firm case study.			There clearly is a psychological component to the effectiveness an ad may have - although in some cases the appeal of the media selected and the surprising creative are major variables in terms of attention and engagement.			admsci6020005_perova	0
+A close reading of some of the cited work (i.e., Mark and Pearson 1991) recognizes that archetypes can be blended.			At a minimum, this exploratory paper should set a more specific and ambitious plan for systematic future research on the topic.			admsci6020005_perova	0
+This was unfortunately not possible in the present iinnovation project.			The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			agriculture12060868_makarova	0
+There are only minor comments with very specific issues:			The following changes could improve the quality of the paper.			agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			There are only minor comments with very specific issues:			agriculture12060868_makarova	0
+As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			However, it would be more elegant to have more batches.			agriculture12060868_makarova	0
+Author Response Figure 1 was missed from the last MS.			The writing and data presenting need to be improved.			agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			This was unfortunately not possible in the present iinnovation project.			agriculture12060868_makarova	0
+The reviewer is completely right in adressing this point.			The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			The writing and data presenting need to be improved.			agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			There are only minor comments with very specific issues:			agriculture12060868_makarova	0
+The reviewer is completely right in adressing this point.			The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+Author Response Figure 1 was missed from the last MS.			The reviewer is completely right in adressing this point.			agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			This was unfortunately not possible in the present iinnovation project.			agriculture12060868_makarova	0
+Finally, thank you for being so patient with our manuscript			Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			agriculture12060868_makarova	0
+The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			agriculture12060868_makarova	0
+Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			The writing and data presenting need to be improved.			agriculture12060868_makarova	0
+As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			As mentioned earlier, we did not have resources t continue the data collection within the time period we had.			agriculture12060868_makarova	0
+There are only minor comments with very specific issues:			The following changes could improve the quality of the paper.			agriculture12060868_makarova	0
+Andersen and Marko have analyzed the design of farrowing pens and present the first production results of the “SowComfort farrowing pen”.			The reviewer is completely right in adressing this point.			agriculture12060868_makarova	0
+Author Response Figure 1 was missed from the last MS.			The paper is clearly written, with nice figures and with a discussion supported by the results obtained.			agriculture12060868_makarova	0
+This was unfortunately not possible in the present iinnovation project.			The paper deals with an interesting topic and totally up to date to provide farmers with solutions for free farrowing (which in Europe may become mandatory under The End of the Cage Age initiative).			agriculture12060868_makarova	0
+line 115: Did one repetition mean one plant?line 130: How many fruits were used as a sample of representative fruits?			Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			agronomy12061433_makarova	0
+It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			Changes in respect to the previous version are in track changes.			agronomy12061433_makarova	0
+There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			2) In figure 3: why CP cultivars in purple lines are clustered with CC and MG in green?			agronomy12061433_makarova	0
+It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			agronomy12061433_makarova	0
+Additional corrections through the text have been done.			See the attachment Comments for author File: Comments.pdf			agronomy12061433_makarova	0
+#Answer: Again, we would like to thank You for your attention and for carefully reading the manuscript.			We also thank you for the positive and encouraging comment.			agronomy12061433_makarova	0
+See the attachment Comments for author File: Comments.pdf			Line 245: would be better to show the value of each agronomic trait mentioned here			agronomy12061433_makarova	0
+Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			agronomy12061433_makarova	0
+Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			line 135: How many fruits were used as a bulk of representative fruits?			agronomy12061433_makarova	0
+We have carefully considered comments and thoughtful suggestions, revising the manuscript accordingly.			This work is quite interesting and is well-written.			agronomy12061433_makarova	0
+We have carefully considered comments and thoughtful suggestions, revising the manuscript accordingly.			Changes in respect to the previous version are in track changes.			agronomy12061433_makarova	0
+There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			agronomy12061433_makarova	0
+Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			Changes in respect to the previous version are in track changes.			agronomy12061433_perova	0
+See the attachment Comments for author File: Comments.pdf			Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			agronomy12061433_perova	0
+This work is quite interesting and is well-written.			This work is quite interesting and is well-written.			agronomy12061433_perova	0
+It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			See the attachment Comments for author File: Comments.pdf			agronomy12061433_perova	0
+There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			agronomy12061433_perova	0
+#Answer: Again, we would like to thank You for your attention and for carefully reading the manuscript.			 The results are described and discussed in detail.			agronomy12061433_perova	0
+1) The introduction part is a bit long, although it states the importance of this local variety of sweet pepper and of tracing its origin.			It presents interesting original results on the assessment of the diversity existing for the sweet pepper local variety ‘Peperone Cornetto di Pontecorvo’.			agronomy12061433_perova	0
+There are just a few minor comments: lines 102 and 113: Were the seeds collected in 2000 and sown in 2021?			line 115: Did one repetition mean one plant?line 130: How many fruits were used as a sample of representative fruits?			agronomy12061433_perova	0
+See the attachment Comments for author File: Comments.pdf			See the attachment Comments for author File: Comments.pdf			agronomy12061433_perova	0
+#Answer: Again, we would like to thank You for your attention and for carefully reading the manuscript.			Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			agronomy12061433_perova	0
+This work is quite interesting and is well-written.			Below point by point response to concerns Reviewer 2 Tripodi et al., seeks to set a basis of authenticity of ‘Peperone Cornetto di Pontecorvo’ sweet pepper by using a combinatory approach of biochemical, genetic, and agronomic markers/traits.			agronomy12061433_perova	0
+ We hope that these revisions successfully address the raised concerns, remaining available for any other requests regarding the manuscript.			 We hope that these revisions successfully address the raised concerns, remaining available for any other requests regarding the manuscript.			agronomy12061433_perova	0
+Author Response Dear Reviewer We would like to thank You for your attention to our work and for carefully reading our manuscript.			#Answer: Again, we would like to thank You for your attention and for carefully reading the manuscript.			agronomy12061433_perova	0
+It is perfectly feasible that some owners did not leave their homes because they were for example, ill, but they did so during the pandemic.			We have addressed your comments and responded in the attached file.			ani12040482_perova	0
+L22 and elsewhere: dogs’ “leaving time” or “leaving hours” I think this expression is misleading because it is not the dogs that have left home, but the owners.			According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			ani12040482_perova	0
+Aa little more mention of issues of SRBs from owner perspective (e.g.			half of the dogs from the SRB group moved to the Clear group in October.			ani12040482_perova	0
+There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing?			Line 131: There was an error with referencing wanted to flag.			ani12040482_perova	0
+I do think this is of interest to a range of stakeholders.			Line 96-97- some more detail on the findings of these various surveys would be beneficial similarly to Bowen's study, just so provide a clear background/discussion of literature setting scene for your study.			ani12040482_perova	0
+You note that the pseudo-R2 was small but I think you could go into more details and note the possibility that it was unrelated too.			This information might be presented in your other paper, but it would be good to include this basic information here as well, so readers can better understand your methods and findings.			ani12040482_perova	0
+Materials and Methods: Line 151: how was the criterion of “at least 5 minutes” chosen?			The sample size is good, and the results are interesting.			ani12040482_perova	0
+If other people were present, did all of them have to be away or just the dog’s owner?			However, I don't think the figures were referred to in the text (unless that was the errors highlighted below?)			ani12040482_perova	0
+I hope this review helps – thank you for the opportunity.			According to the authors, the main finding is that dogs whose leaving time reduced the most during the lockdown were most at risk of developing new SRBs.			ani12040482_perova	0
+Author Response Our replies to these comments are given in italics below each comment.			Impact of the first COVID-19 lockdown on management of pet dogs in the UK.			ani12040482_perova	0
+Larger issues: I would like to see the authors address a few different angles in the discussion 1.			Line 382-384 „To our knowledge, this is the first empirical evidence to show a link between changes in dogs’ leaving routines and risk of SRBs.” The authors take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed.			ani12040482_perova	0
+Study periods: It may be useful to spell out a little more here why these survey periods were chosen here, this is covered elsewhere but think could be made clearer here and earlier.			The whole section is not relevant to the main question.			ani12040482_perova	0
+Impact of the first COVID-19 lockdown on management of pet dogs in the UK.			noise complaints and issues with housing; property damage; decline of human-pet bond) could be beneficial (line 56-57).			ani12040482_perova	0
+As I wrote in the previous manuscript review: “Impact of changes in time left alone on separation-related behaviour in UK pet dogs” is one more paper on the behavior of dogs in the COVID-19 pandemic developed in the last two years.			This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			ani12040482_perova	0
+There would be evidence for some behaviors, such as destructive behavior or defecation, but how would owners know about pacing or tail-chasing?			We have addressed your comments and responded in the attached file.			ani12040482_perova	0
+The article, “Impact of Changes in Owner Leaving Pattern on Separation-Related Behaviour in Pet Dogs in the UK” investigated whether the prevalence of SRB in dogs changed with changes in dog management during and after COVID lockdowns.			I would suggest highlighting and discussing this outcome more.			ani12040482_perova	0
+The sample size is good, and the results are interesting.			This comparison seems essential—if some dogs gets worse but some get better, than the argument that changes due to COVID might bring about new SRB becomes weaker.			ani12040482_perova	0
+Impact of the first COVID-19 lockdown on management of pet dogs in the UK.			Discussion: The results are appropriately discussed though perhaps more discussion about further study would be beneficial.			ani12040482_perova	0
+Line 29: change “dogs whose leaving hours reduce most” to “dogs whose time left alone decreased most” Thank you, this edit has been applied.			Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing.			ani12040482_perova	0
+The authors gathered data (via owner surveys) about the same dogs before the pandemic, at the beginning of the pandemic and approx.			It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g justification for this/why this time duration used.			ani12040482_perova	0
+Can you address the dogs that improved between Feb and Oct?			Conclusions: I believe Animals requires a Conclusions section.			ani12040482_perova	0
+L127-137 The dates of data collections have been already mentioned in L121-126, so there is no need for repeating them.			Results: These were generally well-detailed with clear layout considering scope of results.			ani12040482_perova	0
+L127-137 The dates of data collections have been already mentioned in L121-126, so there is no need for repeating them.			P6 L176 I am assuming you excluded dogs that were not left alone in February (or were there none?)			ani12040482_perova	0
+My only suggestion is that making it clear why February and October were chosen as survey-points may be beneficial.			Results: These were generally well-detailed with clear layout considering scope of results.			ani12040482_perova	0
+Line 366-369= I'm not sure it's totally fair to interpret this (the dog not always bringing toy when greeting them) as an anthropomorphic interpretation on the part of the owner- the anthropomorphic interpretation seems to be from authors of manuscript not owner?			But this is the opinion of a person using English as a second language.			ani12040482_perova	0
+Journal of the American Veterinary Medical Association, 219(4), 460-466.			Can you address the dogs that improved between Feb and Oct?			ani12040482_perova	0
+P6 L176 I am assuming you excluded dogs that were not left alone in February (or were there none?)			This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			ani12040482_perova	0
+Manuscript has been slightly improved, but my comments were not included.			Can you address the dogs that improved between Feb and Oct?			ani12040482_perova	0
+However, the authors do not focus on this result and do not explain this finding.			"The authors have hardly studied all the publications on how dogs behave when daily routines were changed, so the term ""first study"" is exaggerated."			ani12040482_perova	0
+We have addressed your comments and responded in the attached file.			It seems that the authors of this study have already tackled this topic exhaustively in their previous articles			ani12040482_perova	0
+Here are some suggestions that can be made throughout the paper:			In other words, 1407/1807 (78%) of dogs were clear of SRB in February and 90.1% in October.			ani12040482_perova	0
+L127-137 The dates of data collections have been already mentioned in L121-126, so there is no need for repeating them.			Minor issues: Line 35: insert “of” after 9.9%			ani12040482_perova	0
+Risk factors and behaviors associated with separation anxiety in dogs.			It seems that the authors of this study have already tackled this topic exhaustively in their previous articles			ani12040482_perova	0
+That dogs showing SRB prior to lockdown were more likely to show it after.			This is a well written and clear manuscript which would be of interest to academics, practitioners and dog owners.			ani12040482_perova	0
+Therefore, the detailed description of the pandemic is unnecessary and could be moved to the Supplemental material, together with Figure			Introduction: This is clear and in general provides a good background to the topic (bar a couple of points where more expansion would be beneficial).			ani12040482_perova	0
+When looking at SRB status in October, of 1,187 dogs who were left alone in October, 117 (9.9%) were reported to have shown at least one SRB”.			It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g justification for this/why this time duration used.			ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			The authors examined how separation-related behaviors in pet dogs changed in relation to time spent away from home by UK owners in February 2020 (pre-pandemic), May-July 2020 (first lockdown during the pandemic), and October-November 2020 (local tier-system for restrictions; follow-up).			ani12040482_perova	0
+This is good news, and it is unclear for me why is it reported as bad news.			(*Christley, R.M.; Murray, J.K.; Anderson, K.L.; Buckland, E.L.; Casey, R.A.; Harvey, N.D.; Harris, L.; Holland, K.E.; McMillan, 523 K.M.; Mead, R.; et al.			ani12040482_perova	0
+Some more detail on the application of the findings to practitioners/owners would be beneficial.			Animals 2021, 11, 5, 524 doi:10.3390/ani11010005; *Holland, K.E.; Owczarczak-Garstecka, S.C.; Anderson, K.L.; Casey, R.A.; Christley, R.M.; Harris, L.; McMillan, K.M.; Mead, 529 R.; Murray, J.K.; Samet, L.; et al.			ani12040482_perova	0
+I have minor comments below and a few larger comments:			The sample size is good, and the results are interesting.			ani12040482_perova	0
+Line 382-384 „To our knowledge, this is the first empirical evidence to show a link between changes in dogs’ leaving routines and risk of SRBs.” The authors take priority in the empirical demonstration of changes in dogs when the routine of the day is disturbed.			Line 30: change “leaving hours increase again” to “time left alone increased again” Thank you, this edit has been applied.			ani12040482_perova	0
+“More Attention than Usual”: A Thematic Analysis of Dog Ownership Experiences in the 530 UK during the First COVID-19 Lockdown. Anim. 2021, Vol. 11, Page 240 2021, 11, 240, doi:10.3390/ANI11010240).			L314 Table 2: bold is not used here although it was used in Table 1			ani12040482_perova	0
+Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			Discussion: The results are appropriately discussed though perhaps more discussion about further study would be beneficial.			ani12040482_perova	0
+This Figure should be moved to the Supplement material as it tells nothing about the dogs’ behaviour.			The sample size is good, and the results are interesting.			ani12040482_perova	0
+Introduction: This is clear and in general provides a good background to the topic (bar a couple of points where more expansion would be beneficial).			 Line 461: change “onto” to “on to” Thank you, we’ve corrected the typo.			ani12040482_perova	0
+Seems a comparison between SRB- (baseline) → SRB + (post lockdown) and SRB + (baseline) → SRB- (post lockdown) dogs is critical and missing.			However, an explanation would be useful for those readers who are not familiar with this type of depicting.			ani12040482_perova	0
+The sample size is good, and the results are interesting.			But the questionnaire only asked how long the dogs had been alone and what they were doing during this time.			ani12040482_perova	0
+This is a great figure, thank you for including it.			Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+It seems that the authors of this study have already tackled this topic exhaustively in their previous articles			The article, “Impact of Changes in Owner Leaving Pattern on Separation-Related Behaviour in Pet Dogs in the UK” investigated whether the prevalence of SRB in dogs changed with changes in dog management during and after COVID lockdowns.			ani12040482_perova	0
+L148 Please explain why a periof of 7 days were asked to be reported			My main question when reading this section concerned how owners would know that certain separation-related behaviors occurred when they were away?			ani12040482_perova	0
+Line 214: if possible (might not be possible if this was how it was phrased in the survey), change “different to before” to “different from before”			Figure 1- Perhaps just double-check this is referred to in the text.			ani12040482_perova	0
+I do think this is of interest to a range of stakeholders.			The article is generally well-written, the topic is timely, important, and interesting from a dog welfare aspect.			ani12040482_perova	0
+Author Response Our replies to these comments are given in italics below each comment.			The article is generally well-written, the topic is timely, important, and interesting from a dog welfare aspect.			ani12040482_perova	0
+Line 166: define “devolved nations” for readers unfamiliar with the term.			What is surprising though is that 55.7% of the dogs who were reported to show SRB in February (before the pandemic) were clear in October which is good news.			ani12040482_perova	0
+Line 30: change “leaving hours increase again” to “time left alone increased again” Thank you, this edit has been applied.			Results: These were generally well-detailed with clear layout considering scope of results.			ani12040482_perova	0
+It may be beneficial to make it clear why cut off of 'at least 5 mins' was used, e.g justification for this/why this time duration used.			The authors examined how separation-related behaviors in pet dogs changed in relation to time spent away from home by UK owners in February 2020 (pre-pandemic), May-July 2020 (first lockdown during the pandemic), and October-November 2020 (local tier-system for restrictions; follow-up).			ani12040482_perova	0
+“More Attention than Usual”: A Thematic Analysis of Dog Ownership Experiences in the 530 UK during the First COVID-19 Lockdown. Anim. 2021, Vol. 11, Page 240 2021, 11, 240, doi:10.3390/ANI11010240).			In other words, 1407/1807 (78%) of dogs were clear of SRB in February and 90.1% in October.			ani12040482_perova	0
+An error message occurred in several places (e.g., line 131, line 246, line 265)			Materials and Methods: Line 151: how was the criterion of “at least 5 minutes” chosen?			ani12040482_perova	0
+Lines 383-384: Change “dogs’ leaving routines” to “dogs’ time left alone” Thank you, this edit has been applied.			L118 Materials and Methods This section (after the Ethics) should begin with the Subjects section.			ani12040482_perova	0
+This information would have been interesting if data on the mood, stress level, general mental health of the owners had also been collected in connection with the pandemic and the links with dog behaviour have been investigated.			Author Response Thank you for your time in reviewing this manuscript and for your constructive, helpful feedback.			ani12040482_perova	0
+The authors examined how separation-related behaviors in pet dogs changed in relation to time spent away from home by UK owners in February 2020 (pre-pandemic), May-July 2020 (first lockdown during the pandemic), and October-November 2020 (local tier-system for restrictions; follow-up).			However, currently, the paper seems too long, and it goes into too much detail about how the pandemic took place in the UK.			ani12040482_perova	0
+The curve in the plot is not due to a compensation issue, it is demonstrating the up-regulation of CD36, which is followed by up-regulation of CD11b.			The manuscript is well written, the results support the hypotheses.			antiox11040683_makarova	0
+We have included what we have written below for your convenience: “The U937 cell line was originally isolated from the pleural effusion of a patient with histiocytic lymphoma [42].			Authors addressed all my requests and suggestions to my fullest satisfaction.			antiox11040683_makarova	0
+We have included what we have written below for your convenience: “The U937 cell line was originally isolated from the pleural effusion of a patient with histiocytic lymphoma [42].			We agree with your comments and have addressed these to the best of our ability in the revised manuscript.			antiox11040683_makarova	0
+Thus, authors should discuss limitations in interpreting their findings.			The black population illustrates the isotype control for the FITC and PE conjugated antibodies.			antiox11040683_makarova	0
+Taken together, the U937 cell line presented as a good cellular model to study the putative antioxidant effect of the BCAT1 CXXC motif in myeloid differentiation, which is a defining feature of AML [50].			Taken together, the U937 cell line presented as a good cellular model to study the putative antioxidant effect of the BCAT1 CXXC motif in myeloid differentiation, which is a defining feature of AML [50].			antiox11040683_makarova	0
+10 + description) steady state inverstigation is acceptable.			Author Response Thanks for your encouragement and suggestion.			app12052642_perova	0
+[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			Author Response Thanks a lot for your encouragement and suggestion.			app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			Chen, G.; Ma, F.; Hao, G.; Zhu, W. Modeling large deflections of initially curved beams in compliant mechanisms using 277 chained Beam-Constraint-Model, ASME J. Mech.			app12052642_perova	0
+Robot., 2019, 11: 011002, https://doi.org/10.1115/1.4041585.			[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			app12052642_perova	0
+"A theoretical model is derived for the DL-LET joint and verified by FEA and experimental results."""			Author Response Thanks for your encouragement and suggestion.			app12052642_perova	0
+Author Response Thanks a lot for your encouragement and suggestion.			[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			app12052642_perova	0
+10 + description) steady state inverstigation is acceptable.			"At the beginning of the last paragraph of Introduction: ""In this work, based on the design concept of improving the tensile stiffness of LET joints in Ref."			app12052642_perova	0
+Following the reported physical experiments (Fig.			Author Response We thanks the reviewer for the encouragement and valuable comments.			app12052642_perova	0
+The current paper proposes a novel lamina emergent torsional (LET) joint named double-laminated lamina emergent torsional joint (DL-LET) joint which utilizes a layer of flexible H18 aluminum foil to increase the tensile stiffness of LET joint.			Author Response We thanks the reviewer for the encouragement and valuable comments.			app12052642_perova	0
+Following the reported physical experiments (Fig.			Chen, G.; Ma, F.; Hao, G.; Zhu, W. Modeling large deflections of initially curved beams in compliant mechanisms using 277 chained Beam-Constraint-Model, ASME J. Mech.			app12052642_perova	0
+[19], this paper proposed a DL-LET joint by fixing a layer of flexible H18 aluminum foil a LET joint.			Robot., 2019, 11: 011002, https://doi.org/10.1115/1.4041585.			app12052642_perova	0
+Following the reported physical experiments (Fig.			Nice piece of research but one aspect needs to be addressed.			app12052642_perova	0
+Chen, G.; Ma, F.; Hao, G.; Zhu, W. Modeling large deflections of initially curved beams in compliant mechanisms using 277 chained Beam-Constraint-Model, ASME J. Mech.			The flow of the paper or the organization of the paper has to be presented at the end of the introductory section.			app12052642_perova	0
+Author Response We thanks the reviewer for the encouragement and valuable comments.			Author Response Thanks a lot for your encouragement and suggestion.			app12052642_perova	0
+10 + description) steady state inverstigation is acceptable.			Author Response Thanks a lot for your encouragement and suggestion.			app12052642_perova	0
+Following the referee’s suggestion we have added the reference to Mekhov’s paper.			Concerning the question of radiationpressure forces, we take them into account on the atoms but not on the mirrors.			atoms4010002_makarova	0
+(b) There will be heating limiting the available measurement time.			Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.			atoms4010002_makarova	0
+Following the referee’s suggestion we have added the reference to Mekhov’s paper.			I support publication, but I have a few comments that the authors hopefully can address before publication.			atoms4010002_makarova	0
+Author Response The authors present a new interpretation to study Bloch oscillations in a cavity sustained optical lattice and the potential use of this system for very sensitive force measurement.The idea is closely related to previous work as Ref.			Indeed, we point out a possible way forward in the last section that builds on the insight gained from the optomechanics analogy.			atoms4010002_makarova	0
+The sensitivity of their measurements, after many years of development, are only one order of magnitude above our estimated sensitivity, and their measurements take one hour whereas ours should take one second which means it might have applications in rather different situations (e.g.			Nevertheless, the Bloch oscillation frequency is robust against this depth modulation because it only depends on the lattice period, not its depth.”			atoms4010002_makarova	0
+However, a change in the depth of a periodic potential does not change the Bloch frequency because this depends on the lattice’s spatial period, not the depth.			(3) The authors claim there should be no backaction of the field on the oscillation frequencies.			atoms4010002_makarova	0
+(3) The authors claim there should be no backaction of the field on the oscillation frequencies.			To my knowledge this was first discussed in: Gangl, M., EPJD, 8(1), 29-40.			atoms4010002_makarova	0
+(2) It is very helpful to connect cavity output spectra and forces.			Author Response The authors present a new interpretation to study Bloch oscillations in a cavity sustained optical lattice and the potential use of this system for very sensitive force measurement.The idea is closely related to previous work as Ref.			atoms4010002_makarova	0
+Some rough estimate on the time scales should be added.			Hence, in principle I consider it suitable for publication.There are however a few things to be clarified before publication:			atoms4010002_makarova	0
+The auhtors investigate the motion of atoms in a periodic potential inside he cavity taking into account the effect of continuous measurement of photons leaking out of the cavity.			Nevertheless the presentaion sheds new light onto the underlying microscopic dynamics and connects the dynamics to proven optomechanical models of cavity BEC dynamics.			atoms4010002_perova	0
+There are however a few things to be clarified before publication: (1) It should be somehow mentioned more clearly (maybe in the abstract) that the idea is not completely new but work presents a new and more intutive picture of the system dynamics.			Hence, in principle I consider it suitable for publication.			atoms4010002_perova	0
+B: Atomic, Molecular and Optical Physics, 45(10), 102001).			Author Response The authors present a new interpretation to study Bloch oscillations in a cavity sustained optical lattice and the potential use of this system for very sensitive force measurement.The idea is closely related to previous work as Ref.			atoms4010002_perova	0
+This will generate extra noise, but might also help as it reduces atom number uncertainty in the cavity.			Some rough estimate on the time scales should be added.			atoms4010002_perova	0
+Rev. A 88, 033830 (2013). I. B. Mekhov and H. Ritsch, Phys. Rev. Lett. 102, 020403 (2009). J. F. Corney and G. J. Milburn, Phys. Rev. A 58, 2399 (1998). I. D. Leroux, M. H. Schleier-Smith, and V. Vuletić, Phys. Rev. Lett. 104, 073602 (2010).			The idea is closely related to previous work as Ref.			atoms4010002_perova	0
+Rev. A 88, 033830 (2013). I. B. Mekhov and H. Ritsch, Phys. Rev. Lett. 102, 020403 (2009). J. F. Corney and G. J. Milburn, Phys. Rev. A 58, 2399 (1998). I. D. Leroux, M. H. Schleier-Smith, and V. Vuletić, Phys. Rev. Lett. 104, 073602 (2010).			Also the BEC will lead to an effective change of the cavity length and lattice constant, which should renormalize the wavelength and thus om_recoil.			atoms4010002_perova	0
+Hence, in principle I consider it suitable for publication.There are however a few things to be clarified before publication:			Hence, in principle I consider it suitable for publication.There are however a few things to be clarified before publication:			atoms4010002_perova	0
+M. D. Lee, S. Rist, and J. Ruostekoski, New J. Phys. 14, 073057 (2012).			This is somehow in contradiction with theeigenfrequency calculations (see Fig.4) in Horak, P.,Physical Review A, 61(3), 033609, 2000.			atoms4010002_perova	0
+· Line 138: insert “the” before “TDCS” also inserting “even at relativistic energies” would strengthen this intent of this work · Line 139: reference 30 should be replaced with a theoretical reference or the text changed to not imply that ref 30 is a theoretical work · Line 140: delete “one” and insert “the” before “non” plus change “that this would enhance” to “this enhances” Also lines 141-143 are probably meant to be a single sentence.			Also, round the numbers in Fig 6 to whole keV units.			atoms4010010_makarova	0
+The main point in this work is to theoretically study relativistic ionization.			Again, we appreciate all your insightful comments.			atoms4010010_makarova	0
+At least the authors should mention the differences of the two methods and how far the RPWBA can be compared to rDWBA?			-------------------------------------------------------------------------------------------------------------------------- Page 1: * Line 7: it is referenced to the recent theories given in [2] and [3] which are from years 1994 and 2002 respectively.			atoms4010010_makarova	0
+Author Response %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Answer to the third referee’s report of the Manuscript ID atoms-114670 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% The article presents triple differential cross sections (tdcs) calculations of hydrogen ionization by positron and electron impact, in the relativistic and non-relativistic limit, using relativistic plane-wave Born approximation and non relativistic approximations.			Explanation of the differences between the peaks corresponding to the different projectiles is given.			atoms4010010_makarova	0
+The authors did not mention these experimental works in the manuscript.			· Line 129: insert “the” before “TDCS” · Table 2: number notation · Line 134: replace “by” with “with” · Lines 135-137: delete this sentence as it says exactly the same as the previous sentence and Table 2.			atoms4010010_makarova	0
+Also, round the numbers in Fig 6 to whole keV units.			Evidences of this includes (i) some errors in English typing, e.g.			atoms4010010_makarova	0
+Author Response %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Answer to the first referee’s report of the Manuscript ID atoms-114670 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% In the paper, the authors report the theoretical calculation of triple differential cross sections of positron ionization of hydrogen atom at relativistic regime to understand the collision dynamics.			My overall recommendation is that after the listed items are addressed the manuscript be accepted for publication without further review.			atoms4010010_perova	0
+------------------------------------------- Thank you once again for your valuable comments and corrections.			------------------------------------------- Thank you once again for your valuable comments and corrections.			atoms4010010_perova	0
+------------------------------- Answer of authors ------------------------------- - First of all, we would like to thank you very much for this relevant and detailed report.			The following issues need to be addressed: 1)			atoms4010010_perova	0
+I find no major problems with the manuscript, either in content or presentation.			In my opinion, the primary contribution of this work is the prediction that even at relativistic velocities, the TDCS for positron impact is larger than for electron impact and that the difference increases with impact energy.			atoms4010010_perova	0
+------------------------------- 1) Answer of authors -------------------------------			Although no experimental data are available for comparison, the method is well explained and reported to agree with other electron impact calculations.			atoms4010010_perova	0
+Two processes are considered, one occurs during the interaction between the cold atom cloud and the Raman beams, the other occurs during the free evolution of the cold atom cloud in vacuum.			Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+1 Review Report 1 Originality/Novelty This paper treats an important question concerning the measurement of the gravitational constant G by atom interferometry.			The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			atoms4020014_perova	0
+Authors’ response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			atoms4020014_perova	0
+1 Review Report 1 Originality/Novelty This paper treats an important question concerning the measurement of the gravitational constant G by atom interferometry.			The correction phi_Q was not discussed in the previous papers I know on the subject and I think that the paper must be published but, as explained below, important improvements are needed.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			atoms4020014_perova	0
+Author response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			The results obtained seem reasonable, although I have no time to verify all the derivations.			atoms4020014_perova	0
+Author response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			These results are interesting to the people working in the field of atom interferometer, because the test mass are often used in the practical experiment.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+Authors’ response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+I recommend accepting this manuscript, provided the following points are clarified:			Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+The response to these comments and changes to the manuscript in response to these comments is given below: 1.			The response to these comments and changes to the manuscript in response to these comments is given below: 1.			atoms4020014_perova	0
+I recommend accepting this manuscript, provided the following points are clarified:			Review Report 2 In this manuscript, the influences of an external test mass on the phase of the signal of an atom interferometer are studied theoretically.			atoms4020014_perova	0
+Author response We would like to thank the referee for his/her careful reading of the manuscript and the helpful comments.			The treatment involves quantum corrections which are treated by calculating the Wigner representation of the density matrix and I am not aware of the use of such an extensive use of Wigner representation to describe atom interferometry experiments.			atoms4020014_perova	0
+How can the authors ascertain that mannose does not come from mannans in yeast extract?			The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_makarova	0
+Figure 6: Bacterial EPSs are normally composed of repeating units.			They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			biom12060834_makarova	0
+Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.			433, 434, 454, 458, and 507 : First section 3.6 (and its subsections) should be section 3.5.			biom12060834_makarova	0
+528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			Tick label values for glucose and yeast extract are different.			biom12060834_makarova	0
+433, 434, 454, 458, and 507 : First section 3.6 (and its subsections) should be section 3.5.			How can the authors ascertain that mannose does not come from mannans in yeast extract?			biom12060834_makarova	0
+Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			We will answer in the following paragraphs because all comments were related to the same topic.			biom12060834_makarova	0
+Basically, the work was done at a fairly high professional level, the material is presented clearly, well structured, the data obtained are discussed with the previously obtained literature data.			Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			biom12060834_makarova	0
+The evidences for the partial structure given are not convincing.			The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_makarova	0
+In short, they allow to clearly identify the D-mannose α-(1→2) and α-(1→4) linkages in EPS.			These results suggest they are the prevailing units of the EPS backbone making it a branched one.			biom12060834_makarova	0
+Sphingobactan, a new α-mannan exopolysaccharide from Arctic Sphingobacterium sp.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_makarova	0
+Comment 42: l. 683-684: Food-grade oils are not aliphatic and aromatic hydrocarbons.			The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_makarova	0
+Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_makarova	0
+Casillo, A., Fabozzi, A., Russo Krauss, I., Parrilli, E., Biggs, C. I., Gibson, M. I., … Corsaro, M. M. (2021).			Chatterjee, S., Mukhopadhyay, S. K., Gauri, S. S., & Dey, S. (2018).			biom12060834_makarova	0
+The studied EPS showed promising properties to use in the food industry.			528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			biom12060834_makarova	0
+Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			(I was referring to the chemical method by methylation.)			biom12060834_makarova	0
+They deleted erroneous information about mannose residues, but came up with new conclusions about mannose identity and linkages based solely on chemical shift of anomeric proton/carbon.			In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			biom12060834_makarova	0
+The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			Bacterial EPSs are normally composed of repeating units.			biom12060834_makarova	0
+Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			biom12060834_makarova	0
+However, there is no report that the mannan came from yeast extract (DOI: 10.1016/j.ijbiomac.2019.09.139, DOI: 10.1023/B:WIBI.0000033068.45655.2a).			IITKGP-BTPF3 capable of biological response modification.			biom12060834_makarova	0
+According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			According to Figure 7. the activity of the ascorbic acid is higher than the activity of EPS produced by CamB6.			biom12060834_makarova	0
+Response 2: Dear reviewer, thanks for your valuable comments for improving the quality of this paper.			In addition, the yeast mannan have β-(1→4) linkage and the one found in this work have α-linkage type.			biom12060834_makarova	0
+Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			→ Comment 1: The English language was improved, but there are still errors.			biom12060834_makarova	0
+Comment 4: Acceptability for food applications was not discussed.			Units are not specified on the axes labels or in the legend.			biom12060834_makarova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.)			biom12060834_makarova	0
+→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			The information contained in the other 2D spectra was not exploited to provide structural information.			biom12060834_makarova	0
+Manuscript biomolecules-1691308 In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			biom12060834_makarova	0
+Units are not specified on the axes labels or in the legend.			Answer 1: To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript.			biom12060834_makarova	0
+IITKGP-BTPF3 capable of biological response modification.			(I was referring to peak intensities that are not uniform.			biom12060834_makarova	0
+Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			Comment 15: l. 169-170: “thirty” instead of “thirteen”?			biom12060834_makarova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			biom12060834_makarova	0
+The evidences for the partial structure given are not convincing.			Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.			biom12060834_makarova	0
+Answer 1: To better understand EPS chemical structure, further 2D NMR experiments were performed and results are presented inside the manuscript.			Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			biom12060834_makarova	0
+Author Response Please see the attachment to see the responses.			(I was referring to peak intensities that are not uniform.			biom12060834_makarova	0
+Comment 32: l. 480: Why was linkage analysis not performed?			Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			biom12060834_makarova	0
+These results suggest they are the prevailing units of the EPS backbone making it a branched one.			Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			biom12060834_makarova	0
+Comment 14: l. 165: Table 2 where Table 1 expected Answer 14:			Chatterjee, S., Mukhopadhyay, S. K., Gauri, S. S., & Dey, S. (2018).			biom12060834_makarova	0
+Comment 6: EPS vs. EPSs in plural form should be consistent throughout.			Units are not specified on the axes labels or in the legend.			biom12060834_makarova	0
+→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			Comment 1: Unfortunately, its structure was not fully determined.			biom12060834_makarova	0
+Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			50-51: “improvement of rheological” Answer 8:			biom12060834_makarova	0
+It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.			Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			biom12060834_makarova	0
+), as well as two-dimensional NMR spectroscopy.			The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			biom12060834_makarova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			EPC is isolated from a thermal spring of volcanic origin, which contains a set of various elements, including heavy metals.			biom12060834_makarova	0
+TOCSY and HMBC are mentioned in methods, but no results are presented.			Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.			biom12060834_makarova	0
+528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			Comment 3: Optimization of production constitutes an important part not mentioned in the title.			biom12060834_perova	0
+The new spectra emphasize this even more: for example, I count at least 10 H1/H2 cross peaks of different intensities on the COSY.)			528 and 531: “radical” Answer 36: Suggestion has been included in the revised version of the manuscript Comment 37: l. 535 and 536: “activity” Answer 37: Suggestion has been included in the revised version of the manuscript Comment 38: l. 537: No section 3.6.2 Answer 38:			biom12060834_perova	0
+Comment 2: The manuscript is very long: it contains a lot of information and many references not always pertinent to EPS characterization.			Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+Comment 42: l. 683-684: Food-grade oils are not aliphatic and aromatic hydrocarbons.			The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			biom12060834_perova	0
+This EPS was carefully purified according to standard purification protocols for EPS.			Answer 29: As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript Comment 30: l. 442: “C), whereas the” Answer 30: Suggestion has been included in the revised version of the manuscript Comment 31: l. 471, 600, 678, and 718: Reference format is different; corresponding references could be missing in the list.			biom12060834_perova	0
+Comment 43: l. 711: “to be significantly” Answer 43: Suggestion has been included in the revised version of the manuscript Comment 44: l. 725: “possibility of future” Answer 44: Suggestion has been included in the revised version of the manuscript			Basically, the work was done at a fairly high professional level, the material is presented clearly, well structured, the data obtained are discussed with the previously obtained literature data.			biom12060834_perova	0
+These results suggest they are the prevailing units of the EPS backbone making it a branched one.			Probably, to establish its structure, it is necessary to use chemical methods (hydrolysis, methylation, etc.			biom12060834_perova	0
+Comment 4: Acceptability for food applications was not discussed.			(I was referring to the chemical method by methylation.)			biom12060834_perova	0
+There are inconsistencies in singular- plural concordance between subject and verb as well as noun and pronoun.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+(I was referring to the chemical method by methylation.)			→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			biom12060834_perova	0
+Answer 29: As per the suggestion given by the reviewer, section numbering has been checked thoroughly in the revised version of the manuscript Comment 30: l. 442: “C), whereas the” Answer 30: Suggestion has been included in the revised version of the manuscript Comment 31: l. 471, 600, 678, and 718: Reference format is different; corresponding references could be missing in the list.			The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			biom12060834_perova	0
+Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.			Also, β-glucopyranose and β-galactopyranose structures were identified and the acetylation was confirmed.			biom12060834_perova	0
+Articles (mostly definite, but also indefinite) are often missing and sometimes superfluous.			The authors added a COSY spectrum but did not analyze it.			biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			→ Comment 1: The English language was improved, but there are still errors.			biom12060834_perova	0
+→ Comment 2: The following comments were not addressed correctly: Why was linkage analysis not performed?			Manuscript biomolecules-1691308 In this manuscript, the exopolysaccharide from Bacillus haynesii CamB6 was exhaustively characterized.			biom12060834_perova	0
+(I was referring to the chemical method by methylation.)			Comment 1: Unfortunately, its structure was not fully determined.			biom12060834_perova	0
+Figure 6: Bacterial EPSs are normally composed of repeating units.			Figure 6: Bacterial EPSs are normally composed of repeating units.			biom12060834_perova	0
+According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			According to the 1H NMR spectrum, it can be assumed that EPS has a complex branched structure.			biom12060834_perova	0
+The studied EPS showed promising properties to use in the food industry.			These results suggest they are the prevailing units of the EPS backbone making it a branched one.			biom12060834_perova	0
+Units are not specified on the axes labels or in the legend.			The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_perova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			The authors used the RSM based on central composite design technique to maximize the EPS production by B. haynesii, and also conducted a diverse study of the EPS properties.			biom12060834_perova	0
+The studied EPS showed promising properties to use in the food industry.			Tick label values for glucose and yeast extract are different.			biom12060834_perova	0
+Comment 15: l. 169-170: “thirty” instead of “thirteen”?			Tick label values for glucose and yeast extract are different.			biom12060834_perova	0
+Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			Comment 21: l. 284: No section 2.6.3 Answer 21:			biom12060834_perova	0
+Units are not specified on the axes labels or in the legend.			Comment 22: Figure 1: What is the significance of the tick mark labels on the maps?			biom12060834_perova	0
+Answer 25: Suggestion has been included in the revised version of the manuscript Comment 26: Figure 3: Units are not specified on the axes labels or in the legend.			50-51: “improvement of rheological” Answer 8:			biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			Comment 7: Use a consistent abbreviation (l or L) for liter (including milliliter and microliter) throughout.			biom12060834_perova	0
+Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			Comment 19: l. 247: Volume of 0.2 mM ethanolic DPPH solution?			biom12060834_perova	0
+There are inconsistencies in singular- plural concordance between subject and verb as well as noun and pronoun.			481-482: “spectrum” Answer 33: Suggestion has been included in the revised version of the manuscript Comment 34: I.			biom12060834_perova	0
+It is advisable, in my opinion, to provide information on the content of heavy metals in the EPS, since the authors plan to offer this product for the food industry in the future.			TOCSY and HMBC are mentioned in methods, but no results are presented.			biom12060834_perova	0
+Comment 39: l. 581: “Table S1A” Answer 39: Suggestion has been included in the revised version of the manuscript Comment 40: l. 603: “Figure S1” Answer 40: Suggestion has been included in the revised version of the manuscript Comment 41: l. 636: “Glc and Gal” Answer 41:			Comment 4: Acceptability for food applications was not discussed.			biom12060834_perova	0
+This EPS was carefully purified according to standard purification protocols for EPS.			The studied EPS showed promising properties to use in the food industry.			biom12060834_perova	0
+Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			Units are not specified on the axes labels or in the legend.			biom12060834_perova	0
+Answer 25: Suggestion has been included in the revised version of the manuscript Comment 26: Figure 3: Units are not specified on the axes labels or in the legend.			The presented manuscript is related to the study of an exopolysaccharide isolated from the extremophilic microorganism Bacillus haynesii CamB6.			biom12060834_perova	0
+Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			Comment 35: l. 500 : The chemical shift range is more than from 1 to 5.			biom12060834_perova	0
+Comment 28: Figure 4; This figure is redundant as it gives the same information as Figure 3.			Comment 32: l. 480: Why was linkage analysis not performed?			biom12060834_perova	0
+This EPS was carefully purified according to standard purification protocols for EPS.			Comment 17: l. 228: GPC defined on l. 231 (check) Answer 17:			biom12060834_perova	0
+Author Response Please see the attachment for the responses.			Author Response Please see the attachment to see the responses.			biom12060834_perova	0
+Author Response Thank you so much dear reviewer for your positive comments regarding the revised version of our manuscript.			Based on the 1H NMR spectrum, the purity and/or heterogeneity of the EPS is questioned.			biom12060834_perova	0
+In combination with a high yield of polysaccharide from the bacterial mass, this makes it attractive for practical use.			A 2D COSY spectrum would be necessary to confirm the assignments made on the 1D: e.g., H2- Man is not normally found in the range 3.2-3.5 ppm (the statement ll.			biom12060834_perova	0
+Comment 4: Line 526-527: The proposal should be reformulated.			Comment 43: l. 711: “to be significantly” Answer 43: Suggestion has been included in the revised version of the manuscript Comment 44: l. 725: “possibility of future” Answer 44: Suggestion has been included in the revised version of the manuscript			biom12060834_perova	0
+It could be argued that some RNA is hydrolyzed during extraction, but for this to be the case, one should assume that the cells were not well washed after being pelleted by centrifugation, which is not the case.			Author Response Author’s Response: We appreciated the reviewers’ comment.			biomedicines10030600_makarova	0
+https://www.sciencedirect.com/science/article/abs/pii/S0960894X08011086?via%3Dihub			"The authors presented the paper ""Human Serum Proteins and Susceptibility of Acinetobacter baumannii to Cefiderocol: role of iron transport""."			biomedicines10030600_makarova	0
+They actually show that adding these components to the growth medium does not modify the bactericidal nature of the action of the antibiotic.			"The authors presented the paper ""Human Serum Proteins and Susceptibility of Acinetobacter baumannii to Cefiderocol: role of iron transport""."			biomedicines10030600_makarova	0
+Author Response Author’s Response: We appreciate the reviewers’ comment and the suggestion.			Author Response Author’s Response: We appreciated the reviewers’ comment.			biomedicines10030600_makarova	0
+https://www.sciencedirect.com/science/article/abs/pii/S0960894X08011086?via%3Dihub			We agree that confirming that the effect of cefiderocol is bactericidal under the conditions tested is important.			biomedicines10030600_makarova	0
+3) A scheme summarizing how different molecular defects can converge on similar NMDAR-mediated EPSCs would be helpful in the discussion 4) A small commentary on the possible functional significance of the identified mutation properties for the pathogenesis of certain diseases would be interesting.			2) Please add a chart about tau changes in Figure 3.			brainsci12060789_makarova	0
+These findings highlight the complexity of interpreting effects of disease-causing NMDA receptor missense mutations in the context of neuronal function.			Particularly, GluN2B GoF and LoF mutants show similar phenotypes in the presence of GluN1/2A/2B triheteromers, while the LoF mutation, C456Y, shows much less rescue effect than GoF mutation in the absence of both GluN2A and GluN2B.			brainsci12060789_makarova	0
+The results presented are convincing and I only have a few minor comments.			These findings highlight the complexity of interpreting effects of disease-causing NMDA receptor missense mutations in the context of neuronal function.			brainsci12060789_makarova	0
+The abstract is too long, which does not actually abstract the whole content very well.			There are also a number of sentences that are so complicated for easy read.			brainsci12060789_makarova	0
+1) Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative.			In the absence of both native GluN2A and GluN2B subunits, functional incorporation into synaptic NMDA receptors was attenuated for GoF mutants, or almost eliminated for LoF GluN2B mutants.			brainsci12060789_makarova	0
+The authors conclude that the presence of triheteromeric NMDA receptors, and the types of the mutated GluN2 subunit are all important factors that can influence the function of the NMDA variants.			The results presented are convincing and I only have a few minor comments.			brainsci12060789_makarova	0
+Please verify if the time scale is correct in Fig. 3А.			Finally, we show that having a single null Grin2b allele has only a modest effect on NMDA-EPSC decay kinetics.			brainsci12060789_makarova	0
+Mechanistic insight was gained from experiments in HEK293T cells, which revealed that GluN2B GoF mutants slowed deactivation in diheteromeric GluN1/2B, but not triheteromeric GluN1/2A/2B receptors.			The abstract is too long, which does not actually abstract the whole content very well.			brainsci12060789_makarova	0
+Please find below our point-by-point response.			The authors conclude that the presence of triheteromeric NMDA receptors, and the types of the mutated GluN2 subunit are all important factors that can influence the function of the NMDA variants.			brainsci12060789_perova	0
+In the present study, the authors revealed how some mutations from patients with GRIN2B-related disorders result in defective synaptic transmission.			Using molecular replacement experiments in cultured hippocampal slices and functional properties for diheteromeric and triheteromeric NMDA receptors expressed in HEK293T cells, the authors demonstrated effects of GoF and LoF mutations.			brainsci12060789_perova	0
+1) Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative.			Author Response Thank you to the reviewer for their comments and helpful suggestions.			brainsci12060789_perova	0
+There are also a number of sentences that are so complicated for easy read.			Using molecular replacement experiments in cultured hippocampal slices and functional properties for diheteromeric and triheteromeric NMDA receptors expressed in HEK293T cells, the authors demonstrated effects of GoF and LoF mutations.			brainsci12060789_perova	0
+Please find below our point-by-point response.			Please find below our point-by-point response.			brainsci12060789_perova	0
+Even though fig.2aiii shows a comparable relative peak amplitude, which indicates both GOF and LOF rescue NMDA-EPSC in Grin2b-/- neurons, it looks to me that the absolute amplitude in fig.2aii shows a quite big difference in un-transferred groups which are supposed to be comparable, not as consistent as shown in fig.1ci.			The abstract is too long, which does not actually abstract the whole content very well.			brainsci12060789_perova	0
+Please find below our point-by-point response.			1) Since mutations can affect NMDA receptor localization, data on whether AMPA/NMDA ratios changed would be informative.			brainsci12060789_perova	0
+The discussion interprets the results as supporting evidence of exercise-induced hypoalgesia (EIH) in the patients with FM.			The results are clearly reported and adequately discussed.			brainsci6010008_makarova	0
+Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_makarova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			The methodology is sound and the manuscript is well-written.			brainsci6010008_makarova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			We have addressed each concern below and made changes in the manuscript accordingly.			brainsci6010008_makarova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			Author Response We would like to thank the reviewer for their positive and critical review of our work.			brainsci6010008_makarova	0
+I am uncertain why the authors only report the effect sizes of group differences in the first run.			Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_makarova	0
+The results are clearly reported and adequately discussed.			The methodology is sound and the manuscript is well-written.			brainsci6010008_makarova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			The results are clearly reported and adequately discussed.			brainsci6010008_makarova	0
+Following exercise in FM patients, activity was transiently increased in anterior insula and dorsolateral prefrontal cortex (DLPFC) while activity was transienly decreased following rest.			We have addressed each concern below and made changes in the manuscript accordingly.			brainsci6010008_makarova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			The results are clearly reported and adequately discussed.			brainsci6010008_makarova	0
+The methodology is sound and the manuscript is well-written.			However, in the latter case the authors need to address the issue of multiple correlations.			brainsci6010008_makarova	0
+Considering discussion the results in relation to previous studies on exercise and neuroimaging.			Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_makarova	0
+“ Each temperature was applied twice so I assume averages were used, but the authors should clarify.			Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_makarova	0
+“ Each temperature was applied twice so I assume averages were used, but the authors should clarify.			We have addressed each concern below and made changes in the manuscript accordingly.			brainsci6010008_makarova	0
+Considering discussion the results in relation to previous studies on exercise and neuroimaging.			The methodology is sound and the manuscript is well-written.			brainsci6010008_perova	0
+Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			brainsci6010008_perova	0
+In paralell pain sensitivity to heat was decreased by exercise.			Changes in pain sensitivity after exercise vs. quiet rest were significantly correlated with changes in activity in the DLPFC (exercise vs. rest).			brainsci6010008_perova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			This manuscript describes a cohort randomized crossover design comparing cutaneous heat pain ratings and brain activity during heat stimuli after conditions of quiet rest and cycling in patients with a physician-confirmed diagnosis of Fibromyalgia (FM) and age and sex matched controls.			brainsci6010008_perova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			brainsci6010008_perova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			In paralell pain sensitivity to heat was decreased by exercise.			brainsci6010008_perova	0
+Our responses to the reviewer’s specific concerns follow each point and are indented and in red font.			We have addressed each concern below and made changes in the manuscript accordingly.			brainsci6010008_perova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_perova	0
+In table 3, the subheading “Peak X, Y, X” needs correction.			However, in the latter case the authors need to address the issue of multiple correlations.			brainsci6010008_perova	0
+However, in the latter case the authors need to address the issue of multiple correlations.			Author Response We would like to thank the reviewer for their positive and critical review of our work.			brainsci6010008_perova	0
+A reference for the determination that a dosage of antidepressants was “high” should be provided.			We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_perova	0
+This manuscript describes a cohort randomized crossover design comparing cutaneous heat pain ratings and brain activity during heat stimuli after conditions of quiet rest and cycling in patients with a physician-confirmed diagnosis of Fibromyalgia (FM) and age and sex matched controls.			We feel that the manuscript has been significantly improved as a result of the suggested revisions.			brainsci6010008_perova	0
+Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			Elevations should be revised to higher so that readers do not erroneously believe that a pre-scan application of heat stimuli was administered.			brainsci6010008_perova	0
+We have addressed each concern below and made changes in the manuscript accordingly.			Considering discussion the results in relation to previous studies on exercise and neuroimaging.			brainsci6010008_perova	0
+Author Response We would like to thank the reviewer for their positive and critical review of our work.			This study has compared acute effects of exercise and inactivity in fibromyalgia (FM) and pain-free controls on changes in pain and cerebral activity in response to heat.			brainsci6010008_perova	0
+The writing is clear, the analysis is sound, and the presentation is excellent with a few exceptions.			How many were there that fit in this category?			brainsci6010009_makarova	0
+How many were there that fit in this category?			If so, how many and what would happen to the analysis if you removed them?			brainsci6010009_makarova	0
+"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."			The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			brainsci6010009_makarova	0
+On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			Are there better references to substantiate this point?			brainsci6010009_makarova	0
+These results are relevant and important as thimerosal is still included in global vaccines although they have largely been removed from childhood vaccine in the United States.			If so, how many and what would happen to the analysis if you removed them?			brainsci6010009_makarova	0
+This is a straightforward analysis of an important database with profound implications for public health.			This is an important study that deserves to be published.			brainsci6010009_makarova	0
+If so, how many and what would happen to the analysis if you removed them?			"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."			brainsci6010009_perova	0
+The writing is clear, the analysis is sound, and the presentation is excellent with a few exceptions.			This is a straightforward analysis of an important database with profound implications for public health.			brainsci6010009_perova	0
+On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			Can the authors address this point: is this database publicly available?			brainsci6010009_perova	0
+In the exposed and control groups were there other related diseases such as Autism?			On page 11 line 4 you say levels of Hg in the environment and in humans are increasing but your reference #26 doesn't speak to that point.			brainsci6010009_perova	0
+"In table 1, the ""-"" is used to signify absent values but that should be footnoted in the table precisely what this signifies."			If so, how many and what would happen to the analysis if you removed them?			brainsci6010009_perova	0
+Can the authors address this point: is this database publicly available?			The Odds ratios presented are dose dependent and, above 37.5 uG of Hg, they reach over 4.5 which are tremendous associations between an exposure and a disease outcome.			brainsci6010009_perova	0
+A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).			Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.			bs5040518_makarova	0
+That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth.			Round 1: and Author Response The goal of this study was to distinguish subgroups of relationally aggressive females (i.e., reactively aggressive-only subgroup, reactively-and-proactively aggressive-combined subgroup, non-aggressive subgroup), on the basis of nine factors reflecting individual characteristics, peer-related variables and R3 parental control.			bs5040518_makarova	0
+Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.			This work, as with other recent work, makes clear that the most severely aggressive youth tend to engage in high R2 levels of both proactive and reactive aggression and are otherwise also generally more distressed and dysregulated, as compared with youth who are more moderately aggressive and tend to engage in reactive aggression only, and youth who have low levels of aggression.			bs5040518_makarova	0
+Below are a few comments that could hopefully help improve a solid piece of work.			Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.			bs5040518_makarova	0
+For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.			bs5040518_makarova	0
+Given the severe limitations of their cross-sectional design, the authors need to revise their comments with respect to directionality and their use of causality terms throughout the manuscript.			In my view, this manuscript will make a nice contribution to the literature.			bs5040518_makarova	0
+In my view, this manuscript will make a nice contribution to the literature.			First, it is recommended that the authors provide more information about the selection of the cluster solution.			bs5040518_makarova	0
+Round 1: and Author Response The goal of this study was to distinguish subgroups of relationally aggressive females (i.e., reactively aggressive-only subgroup, reactively-and-proactively aggressive-combined subgroup, non-aggressive subgroup), on the basis of nine factors reflecting individual characteristics, peer-related variables and R3 parental control.			Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model.			bs5040518_makarova	0
+In my view, this manuscript will make a nice contribution to the literature.			In particular, and contrary to what the authors seem to believe, it is not possible to know whether differences between the two aggressive groups reflect differences in levels of relational aggression or in type of relational aggression (reactive only vs. combined), given that the combined group is obviously much more aggressive than the reactive-only group.			bs5040518_makarova	0
+Creating groups based on a cluster analysis may have created unnecessary problems.			Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model.			bs5040518_makarova	0
+Below are a few comments that could hopefully help improve a solid piece of work.			Second, the manuscript should be carefully edited as there are a fair number of grammatical and typographical errors, including lack of subject-verb agreement in places (e.g., using “were” and “was” in the same sentence, both referring to the same measure - CU traits), omitted words, and so forth.			bs5040518_makarova	0
+Some measures such as self-reports of peers’ delinquency are also flawed because of a possible projection bias.			For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			bs5040518_perova	0
+For example, how to explain that the combined group is not more delinquent than the reactively aggressive group although they report more CU traits and more delinquent peers, two known factors in regard to delinquent behaviors?			Round 1: Author Response to Reviewer 3 First, some factors (i.e., CU traits) may be predictors whereas others (i.e., delinquency) may be correlates or consequences of relational aggression.			bs5040518_perova	0
+There are a number of grammatical and lexical problems throughout the manuscript that need to be taken care of.			This work, as with other recent work, makes clear that the most severely aggressive youth tend to engage in high R2 levels of both proactive and reactive aggression and are otherwise also generally more distressed and dysregulated, as compared with youth who are more moderately aggressive and tend to engage in reactive aggression only, and youth who have low levels of aggression.			bs5040518_perova	0
+First, it is recommended that the authors provide more information about the selection of the cluster solution.			There are two areas to address to strengthen the paper a bit more.			bs5040518_perova	0
+There are two areas to address to strengthen the paper a bit more.			The study is strengthened by the person-centered approach to data analysis, by the large sample, by focusing on girls who have been much less studied than boys, and by the clear rationale for the study.			bs5040518_perova	0
+These findings are in contrast to earlier work, which assumed that proactive and reactive aggression represented distinct typologies.			Round 1: and Author Response Interesting paper!!			bs5040518_perova	0
+Second, the authors do not mention why they implicitly selected an additive model (i.e., each factor makes a unique and independent contribution) instead of, for example, an interactional model.			That is, no information is available in the present manuscript about alternative cluster solutions from the analysis, for example, the extent to which BIC and the silhouette coefficient differed for other solutions and so forth.			bs5040518_perova	0
+A better strategy would have been to predict each type of aggression while controlling its overlap with the other type (i.e., include reactive aggression when predicting proactive aggression and vice-versa).			The study is consistent with other recently published work in this area in identifying subgroups that are characterized by their severity of aggression.			bs5040518_perova	0
+Round 1: and Author Response Interesting paper!!			Round 1: and Author Response Interesting paper!!			bs5040518_perova	0
+Round 1: and Author Response Interesting paper!!			Overall, this is a nice paper that looks to make a contribution.			bs5040518_perova	0
+There are two areas to address to strengthen the paper a bit more.			Below are a few comments that could hopefully help improve a solid piece of work.			bs5040518_perova	0
+Below are a few comments that could hopefully help improve a solid piece of work.			Despite the use of a relatively large sample and of sound measures, this study falls short of making an important contribution to the current literature for several reasons.			bs5040518_perova	0
+However, my main concern with the study is that there were too many analyses presented to interpret in a meaningful way.			Participants completed two waves of data collection approximately 1-year apart.			bs6010004_makarova	0
+Participants completed two waves of data collection approximately 1-year apart.			On p. 4 the first sentence of second full paragraph, “Regarding sexual subjectivity and its link to greater sexual exploration and experience, one cross-sectional with a sample of females” is missing the word “study.” The authors could provide more theoretical explanation of why there might be gender differences in sexual subjectivity.			bs6010004_makarova	0
+There is an increase in Type I error with the multiple t-tests conducted thatshould be addressed.			Also, a more thorough description of the measurement of the construct and the validity of the measurement could be described in that section.			bs6010004_makarova	0
+Participants completed two waves of data collection approximately 1-year apart.			**We also expanded the first paragraph of the Limitations to say: Although this study provided insight into how aspects of sexual subjectivity differed over one year in young men and women and uncovered associations of age and sexual behavior with sexual subjectivity, there were two limitations worthy of note.			bs6010004_makarova	0
+"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion."			**We did not keep an exact count of the number of students approached about the study, but we estimate that we approached about 375 students.			bs6010004_makarova	0
+Although an exact count was not available, we estimate that 375 students were approached to participate, resulting in a participation rate of approximately 79%.			Using a longitudinal design with two assessments, we investigated patterns of entitlement to sexual partner pleasure and self-efficacy to achieve sexual pleasure among 295 young men and women aged 17-25 years attending one Australian university.			bs6010004_makarova	0
+That seems to be the model implied by the analyses, but it is never explicitly specified.			** We added a comment that data were collected at one Australian university to the abstract, and added a summary statement and note about limitations related to generalizability.			bs6010004_makarova	0
+We also tested whether entitlement and efficacy differed by gender, and hypothesized that entitlement and efficacy would be higher in older participants and those with more sexual experience.			"Finally, on a selfish note, I wrote an extensive review piece on this topic (Harden, 2014, ""A Sex-Positive Framework for Reseach on Adolescent Sexuality"" in Perspectives on Psychological Science), and I would, of course, be happy if the authors included a reference to my paper in their Introduction or Discussion."			bs6010004_makarova	0
+Important limitations of the study including that this was a study of students at one university only should be included in the Abstract.			At T1, 225 of the participants (76%) reported a history of sexual intercourse and 9% reported having had no sexual experience apart from kissing.			bs6010004_makarova	0
+Moreover there is a comment in the Methods that 80% of the subjects reported being only attracted to members of the same sex.			Why would behavior that has already happened contribute to a re-ordering of individuals over the course of the next year?			bs6010004_makarova	0
+Some more specific suggestions are provided below.			From the relatively high percentage of non-heterosexual participants, there seems to be some selection bias.			bs6010004_makarova	0
+A low response rate will seriously limit the validity of the findings and this should be discussed as an important limitation also.			Moreover there is a comment in the Methods that 80% of the subjects reported being only attracted to members of the same sex.			bs6010004_makarova	0
+There might be some interesting data collected in the present study.			Using a longitudinal design with two assessments, we investigated patterns of entitlement to sexual partner pleasure and self-efficacy to achieve sexual pleasure among 295 young men and women aged 17-25 years attending one Australian university.			bs6010004_makarova	0
+This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.			The authors should revise the paper providing more theoretical background and accompanying analyses.			bs6010004_makarova	0
+The reporting of group differences in the participants section might be better placed in the results section after the measures are described.			With better theoretical framing and a more limited set of analyses, the value of the study results should be more clear.			bs6010004_makarova	0
+However, my main concern with the study is that there were too many analyses presented to interpret in a meaningful way.			We also tested whether entitlement and efficacy differed by gender, and hypothesized that entitlement and efficacy would be higher in older participants and those with more sexual experience.			bs6010004_makarova	0
+There is also the possibility that individuals more interested in sex and relationships chose to participate.			Predictions about how the specific subscales of the inventory might relate to age and experience might be provided to make the results more meaningful (see the comment about theory relating to analyses discussed above).			bs6010004_makarova	0
+That is, any one who did not complete both assessments was not included in the analyses.			What were participants told about the purpose of the study?			bs6010004_makarova	0
+A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			Author Response There might be some interesting data collected in the present study.			bs6010004_perova	0
+Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.			This contradiction underscores my concern about attrition (point #1); I wonder if the seeming decrease from Time 1 to Time 2 is an artifact of attrition.			bs6010004_perova	0
+The combining of the male and female versions of the sexual subjectivity inventories should be described in more detail, and the validity of creating the subscales should be explained.			Third, although longitudinal data can be a quite powerful tool for understanding change and development, the current analyses squander some of that power.			bs6010004_perova	0
+This seems to contrast with later statements and would also seem to be an unlikely finding.			Perhaps factor analysis could be used to determine appropriate subscales?			bs6010004_perova	0
+"Overall, 91% were white/Caucasian, 3% were Asian, 1 was Aboriginal/Pacific Islander, and the remaining participants indicated an ""Other"" sociocultural background."			The reporting of group differences in the participants section might be better placed in the results section after the measures are described.			bs6010004_perova	0
+The authors should revise the paper providing more theoretical background and accompanying analyses.			The abstract now reads: Abstract: Many scholars have called for an increased focus on positive aspects of sexual health and sexuality.			bs6010004_perova	0
+At the very least, the Discussion should consider how generalizable these results are.			Although an exact count was not available, we estimate that 375 students were approached to participate, resulting in a participation rate of approximately 79%.			bs6010004_perova	0
+Please note - we have also shortened the title, found some other typographical errors to fix, and fixed some references.			A sense of entitlement to sexual partner pleasure increased significantly over the year of the study, whereas, on average, there was no change in self-efficacy over time.			bs6010004_perova	0
+**We did not keep an exact count of the number of students approached about the study, but we estimate that we approached about 375 students.			That seems to be the model implied by the analyses, but it is never explicitly specified.			bs6010004_perova	0
+Therefore, in order to capture sexual subjectivity as it emerges and develops most rapidly along with sexual behavior, future studies may need to begin with a younger population.			Please note - we have also shortened the title, found some other typographical errors to fix, and fixed some references.			bs6010004_perova	0
+The abstract now reads: Abstract: Many scholars have called for an increased focus on positive aspects of sexual health and sexuality.			Fifth, one set of results seems quite contradictory: sense of entitlement to sexual self-pleasure decreased, on average, from Time 1 to Time 2, but sense of entitlement to sexual self-pleasure was positively correlated with age.			bs6010004_perova	0
+Most lived with their parents (61%) and reported being only attracted to the other sex (80%).			The abstract now reads: Abstract: Many scholars have called for an increased focus on positive aspects of sexual health and sexuality.			bs6010004_perova	0
+There was no evidence that any association differed between young men and young women.			What percentage of people who were approached agreed to participate?			bs6010004_perova	0
+The authors should revise the paper providing more theoretical background and accompanying analyses.			Perhaps factor analysis could be used to determine appropriate subscales?			bs6010004_perova	0
+Some more specific suggestions are provided below.			Age was positively associated with T1 entitlement, and experience with a wider range of partnered sexual behaviors was concurrently associated with more entitlement and efficacy and was also associated with increased entitlement to partner pleasure and increased self-efficacy in achieving sexual pleasure at T2 relative to T1.			bs6010004_perova	0
+Third, although longitudinal data can be a quite powerful tool for understanding change and development, the current analyses squander some of that power.			Similarly, there are many correlation tests performed which increases Type I error that needs to be addressed.			bs6010004_perova	0
+It seems that a more interesting and direct test of the longitudinal effects of sexual behavior on sexual subjectivity would test whether new sexual behaviors (that is, sexual behaviors experienced between Time 1 and Time 2) predict change in sexual subjectivity from Time 1 to Time 2.			The reporting of group differences in the participants section might be better placed in the results section after the measures are described.			bs6010004_perova	0
+International Journal of Environmental Research and Public Health.			Response 9: The supplemental Information (i.e., office temperature, number of employees) are added in the revised version.			buildings12040417_makarova	0
+Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018.			1， Figure 1 should be improved regarding the arrow.			buildings12040417_makarova	0
+Response 17: Spearman’s correlation test was conducted, and the revised contents are shown in the updated version.			Point 2: The illustrations in the article are small and a bit vague, some pictures can shrink a little, not to the top to the border, these can be optimized.			buildings12040417_makarova	0
+Mollazadeh, M., Zhu, YM., Application of Virtual Environments for Biophilic Design: A Critical Review.			The questionnaire results reveal that the employees from the understudy companies hold a relatively positive opinion on wellbeing, nature-relatedness, indoor environmental quality, and biophilic design for their health promotion.			buildings12040417_makarova	0
+Response 9: The supplemental Information (i.e., office temperature, number of employees) are added in the revised version.			In terms of the reviewer think that the listed attributes span across different domains and the communal features are not that apparent, we believe it is due to the research perspective of biophilic design is different from the perspectives of building science and traditional POE studies (please find the detailed explanations in the respond for Comment #9).			buildings12040417_makarova	0
+Response to Reviewer 3 Comments (Round 2) Buildings-1613867.			Concerning the self-related evaluation scales GH and NR, the score range of GH and HR are minimum 4 to maximum 20, and the obtained results illustrate moderately high opinions, the values of 15.00 (2.00) and 14.00 (3.00) for GH and NR.			buildings12040417_makarova	0
+After reviewing their revised manuscript, I felt that their responses were on-point and addressed my earlier thoughts.			Prospect and refuge Prospect: long views of surrounding settings Refuge: sites of safety and security 20.			buildings12040417_makarova	0
+Similarly, the Green Mark system uses the green plot ratio, assigning credits to greenery provision to enhance biodiversity and visual relief.			The detailed description of the demographic information is in Section 3.1.			buildings12040417_makarova	0
+Practical methodologies to establish biophilic design frameworks for specific workplace typology.			Response to Reviewer 2 Comments Point 1: The study about the evaluation of biophilic attributes in the workplace for improving health and wellbeing is actual and interesting and therefore the paper is fulfilling the scientific criteria to be published.			buildings12040417_makarova	0
+Complexity & Order adheres to a spatial hierarchy that similar to those encountered in nature 11.			Light Glass walls and clerestories, reflecting colors and materials 2.			buildings12040417_makarova	0
+Transitional spaces hallways, thresholds, doorways, gateways, and areas that link the indoors and outdoors especially porches, patios, courtyards, colonnades 23.			Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].” Point 6:			buildings12040417_makarova	0
+A data-driven analysis of occupant workspace dissatisfaction.			There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays.			buildings12040417_makarova	0
+Practical methodologies to establish biophilic design frameworks for specific workplace typology.			Although I appreciated the thoroughness to which the descriptive statistical was explained, I wasn’t convinced the mean was the best indicator for the data, considering that evaluation scores were collected on a 5-point scale and not a continuous linear one.			buildings12040417_makarova	0
+An image showing the indoor conditions and outdoor façade for each might be beneficial.			Response 9: The supplemental Information (i.e., office temperature, number of employees) are added in the revised version.			buildings12040417_makarova	0
+And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version.			There were two very minor points that I came across, which they may want to reconsider before proceeding with their article.			buildings12040417_makarova	0
+Studies proofed that biophilic design benefits workers’ health and productivity in an office environment [60, 61, 62, 63].” Citations are added in the Reference list: 59.			Risk/Peril An identifiable able threat coupled with a reliable safeguard.			buildings12040417_makarova	0
+Complexity & Order adheres to a spatial hierarchy that similar to those encountered in nature 11.			c) Furthermore, the questionnaire results enhance our knowledge on the practical ap-plication of biophilic design frameworks for the workplace and contributed to more framework design consideration.			buildings12040417_makarova	0
+First, to simplify the image, we put all the definitions into a new table in the appendix (Appendix A. Definitions of biophilic design attributes and patterns).			For instance, in most cases, the employers would not create an office environment that makes the workers feel “Prospect”, “Refuge”, “Mystery”, and “Risk”.			buildings12040417_makarova	0
+Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).			And the corresponding figures (Figure 1 in Page 13 Line 359-361 and Figure 4 in Page 16-17 of the original manuscript) are removed in the revised version.			buildings12040417_makarova	0
+The investigation explains that combined literature review and POE results are one of the practical methodologies to establish biophilic design frameworks for specific workplace typology.			Additionally, the effectiveness of such design in practical design projects for user wellbeing still requires confirmation.			buildings12040417_makarova	0
+Physiological and cognitive performance of exposure to biophilic indoor environment.			The same parameters (e.g., daylight, thermal comfort, and air-quality, office layout and building form) can be classified in different classifications (i.e., the traditional POE frameworks and the biophilic design frameworks) by different perspectives.			buildings12040417_makarova	0
+The Cronbach’s Alpha with an Alpha>0.6 considered acceptable internal consistency in this study.			Yin, J., Zhua, S., MacNaughton, P., Joseph, G., Allen, J.G., Spengler, J.D., 2018.			buildings12040417_makarova	0
+), Innovative Developments in Architecture, Engineering and Construction.			Nevertheless, only 47.8% of them selected agreed/strongly agree regarding the statements that their ideal spot for vacation would be a wilderness area (NR1-Q12).			buildings12040417_makarova	0
+The sentence was rewritten to demonstrate this argument.			Intercorrelations between responses of three subscales.			buildings12040417_makarova	0
+d) The correlation results support the importance of biophilic design from the user perspectives.			However, further research on developing building typology-based biophilic design guide-lines and assessment methods are necessary.			buildings12040417_makarova	0
+Although this I don’t believe this would be a large undertaking, the analysis would be better served by non-parametric tests due to the level of measurement used (i.e.			There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace.			buildings12040417_makarova	0
+d) The correlation results support the importance of biophilic design from the user perspectives.			There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace.			buildings12040417_makarova	0
+Response 16: Thanks for the reminder, the error has been corrected in the revised version.			Biophilia: Does Visual Contact with Nature Impact on Health and Well-Being?			buildings12040417_makarova	0
+Effects of biophilic indoor environment on stress and anxiety recovery: a between-subjects experiment in virtual reality.			This would make it easier for the reader; 2) Round the percentages to the nearest whole number.			buildings12040417_makarova	0
+This led me to believe that this office was much larger than the building studied in China.			Cheung, T., Schiavon, S., Graham, L.T., Tham, K.W., 2021.			buildings12040417_makarova	0
+Revision in Page 15-17 Line 399-433: The analysis of individual items provides more details into the works' responses.			In the future study, the research scope should be narrow down for intensive investigation.			buildings12040417_makarova	0
+b) Additionally, the study provides novel design guidelines for designers with emphasizing on weight for workplace design practices.			In-lieu of the mean, please consider using the median and inter-quartile range as the central tendency and dispersion indicators.			buildings12040417_makarova	0
+"Revision in Page 1 Line 28:“The term ""Biophilia"" is evolved from human evolution research and is coined to de-scribe humans' inherent love affinity for the living things in the natural world [1,2].” Point 2: P1, L39-41: Please provide references to these frameworks."			While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.			buildings12040417_makarova	0
+Physiological and cognitive performance of exposure to biophilic indoor environment.			These are exploited to show a new biophilic design framework for the workplace according to the users’ points of view (based on the POE results).			buildings12040417_perova	0
+There are overlaps between the nine biophilic design attributes and these eight influential factors for the workplace.			Both classifications validated by previous literatures.			buildings12040417_perova	0
+The median (IQR) value of the POE scale BDE is evaluated as 42.00 (5.00) (range of total value: min.			Environmental Quality and the Productive Workplace.			buildings12040417_perova	0
+2) However, these patterns are not common in most offices.			Hence, all the means (SD) are replaced as medians (IQR).			buildings12040417_perova	0
+In my view, POE information had more utility diagnosing operation problems, which can be solved when running the building, identifying prominent sources of dissatisfaction that can prompt action to resolve these issues.			Revision in Page 3 Line 109: “It has helped experts to obtain user’s feedbacks over the last five decades [31,32].” Point 6: P3, L106: While I generally agree with, questions could be raised to whether POE scales should be used to evaluate biophilic design evaluation.			buildings12040417_perova	0
+Response 6: The different genders and different ages are included in the study.			As mentioned in the Conclusion, in the future study, we will include more offices and locations as experiment samples.			buildings12040417_perova	0
+how satisfied are you with the biophilic features) may not accurately depict every beneficial nuance they offer (e.g.			According to the research objectives of this study (i.e., evaluate the subjective health impacts of biophilic design in workplace), we need to refer the well-developed scales from other disciplinary (e.g., Environmental Psychology).			buildings12040417_perova	0
+25,39], and the 14 patterns of biophilic design [ref.40].			A quantitative study for indoor workplace biophilic design to improve health and productivity performance.			buildings12040417_perova	0
+Perhaps this makes this article completer and more credible.			And the questionnaire can be applied in future biophilic design research for investigation.			buildings12040417_perova	0
+I think psychological factors will also affect human physiological comfort.			across subsets, leading to no significant differences across subsets.			buildings12040417_perova	0
+Dynamic & Diffuse Light Leveraging varying intensities of light and shadow that change over time to create conditions that occur in nature.			"The introduction of the term ""Biophilia"" can be more detailed and easier to understand."			buildings12040417_perova	0
+Revision in Page 13 Line 381-394: The Cronbach’s α coefficient value of the main scale is 0.72, while those of the subscales GH, NR, and BDE in order are obtained as 0.68, 0.79, and 0.63, indicating that the questionnaire is reliable (i.e., an acceptable reliability: Cronbach's α>0.6) [57, 58] (Table 8).			There are general biophilic design frameworks (e.g., the 24 Biophilic Design Attributes and the 14 Patterns of Biophilic Design) and green building, healthy building standards (e.g., WELL v2 and Singapore Green Mark) that include biophilia into the certification systems in nowadays.			buildings12040417_perova	0
+Both classifications validated by previous literatures.			), Innovative Developments in Architecture, Engineering and Construction.			buildings12040417_perova	0
+outside of POE studies) have been adopted for this reason, and a short sentence explaining the rationale supporting the lack of scales for biophilic design could be provided to briefly mentioned this.			More than 60 percent (approximately 60.7%) of respondents believe that greenery is a biophilic design that benefits office wellbeing (BDE6-Q21).			buildings12040417_perova	0
+According to the quantitative results presented in Table 8, the medians (Interquartile Range, IQR) of the assessment show moderately high opinions toward the health and wellbeing of biophilic offices (HWBO), at 71.00 (8.00) (the score range from min. 20 to max. 100).			Point 3: P2, 47-53: Although in the past there were few guidelines, nowadays, there may be more standards that focus on nature integration within the built environment.			buildings12040417_perova	0
+Author Response Point 1: The introduction can be optimized appropriately.			Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design.			buildings12040417_perova	0
+Hence, the questions in the final section are designed based on the selection of the biophilic design elements/attributes that typically applied in the office design, which are not mentioned in the previous scales.			Revision in Page 6 Line 215-220: “Table 2 made a comparison between the selected nine biophilic design attributes for workplace in this study and the validated eight factors that affect workers’ satisfaction and productivity.			buildings12040417_perova	0
+Further studies could include more offices and locations as experiment samples.			Point 7: Something I felt would useful would at the beginning would be a clear definition for what “biophilic attributes” refers to.			buildings12040417_perova	0
+Point 4: The result of biophilic design frameworks for specific workplace typology is of certain value.			While this is somewhat accurate, it might be more appropriate to elucidate this as an “inherent affinity”.			buildings12040417_perova	0
+The Conclusion is rewritten to highlight the contribution and implementation of the results: Revision in Page 23 Line 516-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing.			#2: P1, L39-41: Please provide references to these frameworks.			buildings12040417_perova	0
+As mentioned in the Conclusion, in the future study, we will include more offices and locations as experiment samples.			9. material Connection with nature Material and elements from nature that, through minimal processing, reflect the local ecology or geology to create a distinct sense of place.			buildings12040417_perova	0
+Additionally, the effectiveness of such design in practical design projects for user wellbeing still requires confirmation.			Questionnaire Results focus on illustrating the questionnaire results.			buildings12040417_perova	0
+Therefore, the weighting results of this experiment are not employed to deny the ranking in the 14 Patterns of Biophilic Design.			Response 6: The different genders and different ages are included in the study.			buildings12040417_perova	0
+Mollazadeh, M., Zhu, YM., Application of Virtual Environments for Biophilic Design: A Critical Review.			Stacked graph with percentage responses for individually arranged items (from the bottom with a high percentage of disagreement to the top with high percentage of agreement).			buildings12040417_perova	0
+If this was the case, then please better articulate its overarching utility in this study, considering that half of the patterns were not relevant to the research scope.			Response 19: The Conclusion is rewritten to highlight the relationship between biophilic design and occupant health and wellbeing: Revision in Page 23 Line 514-540: “The significant research outputs from the present scrutiny are shown as following: a) The authors develop a POE questionnaire for evaluating the biophilic design for workplace health and wellbeing.			buildings12040417_perova	0
+Observation Results-Biophilic Design Attributes in the Selected Offices is moved to be Section 2.3.2.			Practical methodologies to establish biophilic design frameworks for specific workplace typology.			buildings12040417_perova	0
+Response 5: The sentence was rewritten in the updated version.			Although this I don’t believe this would be a large undertaking, the analysis would be better served by non-parametric tests due to the level of measurement used (i.e.			buildings12040417_perova	0
+Second, the study will provide a new biophilic design guidelines for workplaces, which can effectively assist researchers and designers to improve office biophilic design practices and decision-making on design attributes selection.			#2: P1, L39-41: Please provide references to these frameworks.			buildings12040417_perova	0
+The major scale of the questionnaire consists of three parts (subscales): general health (GH), nature relatedness (NR), and biophilic design evaluation (BDE).			This becomes an issue later, since some aspects referring to biophilic design become unclear.			buildings12040417_perova	0
+That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or).			As we all know surgery of chordomas, especially of recurrent chordomas, is challenging and the authors should be congratulated for their thoroughly surgical planning and approaches, using elaborate pre- and intraoperative imaging, which is well described throughout the manuscript.			cancers14040966_makarova	0
+All that is needed is easy intra-operative access to CT and MR imaging and the ability to import and reconstruct the data real time.			The authors are to be commended for pursing this approach in a small series of patients with difficult diagnoses.			cancers14040966_makarova	0
+What means Volume reduction 0 and 0% in Table 4?			However, the PET scan does not appear to have actually been used for intraoperative repeat imaging to assess resection, and fluoroscopy and 3D reconstructions using Brain Lab software are standard fare in many operating rooms.			cancers14040966_makarova	0
+There is not much to add and publication can be recommended.			3D reconstructions of newly acquired data is a capability of the software, and is available in many places.			cancers14040966_makarova	0
+Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions.			Making this clear is important for others who either use the technique or who want to do so.			cancers14040966_makarova	0
+However, patients’ symptoms, tumour location, surgical approach, imaging, complications and outcome are well documented in the tables, and additionally illustrated by two case reports.			As we all know surgery of chordomas, especially of recurrent chordomas, is challenging and the authors should be congratulated for their thoroughly surgical planning and approaches, using elaborate pre- and intraoperative imaging, which is well described throughout the manuscript.			cancers14040966_perova	0
+Intraoperative angiography might have been used in the case of vascular injury but this also is available in many institutions.			That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or).			cancers14040966_perova	0
+As we all know surgery of chordomas, especially of recurrent chordomas, is challenging and the authors should be congratulated for their thoroughly surgical planning and approaches, using elaborate pre- and intraoperative imaging, which is well described throughout the manuscript.			That is plenty and a challenge for many, but the other imaging modalities used are either standard fare in many centers (x-ray imaging, endoscopy) or do not need to be in adjacent rooms (PET scanning and angiography (can be done in the primary Or).			cancers14040966_perova	0
+Muliti-modality intraoperative imaging is an important adjunct for the neurosurgeon and in this regard, the paper is valuable.			However, chordoma is a rare disease and reports on large patient series are rare.			cancers14040966_perova	0
+If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			Comments to authors: Major: Based on the cell line RNA and protein data the expression level of MACC1 is highly variable rather than being present or not.			cancers14071773_makarova	0
+The results are interesting and may have clinical relevance.			The concentration used from selumetinib both for the cell line experiments (10 microM) and for the animal experiments (50 mg/kg) are quite high.			cancers14071773_makarova	0
+Based on these comments we herewith respond to all questions raised in a point-by-point manner, as follows: Reviewer 1: 1.			[2] Abstract says 266 patients were analyzed (line 36), while result says 360 patients were analyzed (line 249).			cancers14071773_makarova	0
+In this paper the authors represents that MACC1 is a negative prognostic marker in esophageal and gastric adenocarcinomas in a large Caucasian cohort.			The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models.			cancers14071773_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			cancers14071773_makarova	0
+Answer: We thank the reviewer for this comment.			It would be nice to know the selumetinib sensitivity of the two used cell lines.			cancers14071773_makarova	0
+FLO1 EV FLO1 MACC1 0 50 100 150 AUC (72h) Percent of Controll MACC1 related Proliferation over 72h p = 0.113 p=0.113			I recommend the following edits before publication.			cancers14071773_makarova	0
+[3] Throughout the paper, 10^x wrongly displays as 10x (e.g.			The authors´answer regarding the cut off is sufficient, please include this argument either in the methods section or as supplementary information.			cancers14071773_makarova	0
+I recommend the following edits before publication.			In the pdf I reviewed, the caption is there, but the figure is not.			cancers14071773_makarova	0
+[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion.			[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).			cancers14071773_makarova	0
+Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?			In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.			cancers14071773_makarova	0
+Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?			Please provide the distribution of the IRS scores of the patient cohort and explain how was IRS 5 chosen as cut off?			cancers14071773_makarova	0
+If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			In the pdf I reviewed, the caption is there, but the figure is not.			cancers14071773_makarova	0
+In the pdf I reviewed, the caption is there, but the figure is not.			Dear Professor Mok, We appreciate that you give us the possibility to revise our manuscript and we thank the reviewers again for their valuable comments.			cancers14071773_makarova	0
+They found that MACC1 expression does not influence cell proliferation but is correlated with cell migration.			"""MACC1 is an independent negative prognostic marker in AGE/S patients."""			cancers14071773_makarova	0
+[7] In line 272/273: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values), (iii) please report both overall survival result and disease-specific survival result (since you showed both in Figures).			Is there a similar variability present among the tissue samples?			cancers14071773_makarova	0
+However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours.			If the difference is statistically significant, please briefly explain/speculate why.			cancers14071773_perova	0
+FLO1 EV FLO1 MACC1 0 50 100 150 AUC (72h) Percent of Controll MACC1 related Proliferation over 72h p = 0.113 p=0.113			It would be nice to know the selumetinib sensitivity of the two used cell lines.			cancers14071773_perova	0
+Inhibition of MEK1 with selumetinib also reduced cell migration only in MACC1 expressing cells.			They demonstrate that MACC1 is expressed in 79% of the investigated samples and its presence is coupled with reduced overall survival even in patients with good prognosis.			cancers14071773_perova	0
+The results are interesting and may have clinical relevance.			"The authors refer to a data ""Proliferation of FLO1 EV and FLO1 MACC1 over 72 h"", however I couldn´t find the figure either in the manuscript file or here in the answer."			cancers14071773_perova	0
+In the in vitro analysis they investigated MACC1 expression in five cell lines and generated a cell line with MACC1 overexpression and a MACC1 knock-down cell line.			In this study, the authors have shown that the same is also true for gastric/esophageal cancer.			cancers14071773_perova	0
+The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models.			"and before ""To analyze the role of MACC1 in vitro the cell lines FLO-1 and OE33 were lentivirally manipulated."""			cancers14071773_perova	0
+Please discuss how the applied concentrations are related to the clinically used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.			However, cell proliferation was analyzed only in a 24-hour long period even though most cancer cells have a doubling time around 24 hours.			cancers14071773_perova	0
+[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			[9] In line 277-279: (i) please report median survival in each group (instead of mean survival), (ii) please specify Hazard ratio with 95% confidence interval (in addition to p values).			cancers14071773_perova	0
+If the journal requires an unstructured abstract instead, please reorganize the current abstract so that the content flows uninterrupted (e.g.			Answer: We thank the reviewer for this comment.			cancers14071773_perova	0
+[3] Throughout the paper, 10^x wrongly displays as 10x (e.g.			[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion.			cancers14071773_perova	0
+Dear Professor Mok, We appreciate that you give us the possibility to revise our manuscript and we thank the reviewers again for their valuable comments.			Inhibition of MEK1 with selumetinib also reduced cell migration only in MACC1 expressing cells.			cancers14071773_perova	0
+It would be nice to know the selumetinib sensitivity of the two used cell lines.			[10] If the journal allows it, please break up the abstract into Introduction, Methods, Results, Conclusion.			cancers14071773_perova	0
+Author Response Dear Professor Mok, We appreciate that you give us the possibility to revise our manuscript and we thank the reviewers for their valuable comments.			[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			cancers14071773_perova	0
+In an in vivo xenograft model using a control and a MACC1 overexpressing cell line pair they found that MACC1 expression increased metastasis formation and this was reduced by selumetinib treatment.			The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models.			cancers14071773_perova	0
+In order to draw a conclusion about the proliferation effect a longer (48 or 72 hours) measurement is necessary.			[5] If possible, please use a higher resolution version of Figures 3F and 3I (the legends are blurry and hard to read in the current version).			cancers14071773_perova	0
+Please discuss how the applied concentrations are related to the clinically used dosage (75 mg administered orally twice https://clinicaltrials.gov/ct2/show/NCT02448290) and why not the more efficient MEK1 inhibitor trametinib was used for the experiments.			The authors argue that MACC1 expression did not significantly influenced cell proliferation in the two cell line models.			cancers14071773_perova	0
+The materials and methods section are elaborated in the details.			ANOVA = analysis of variance; HCC = hepatocellular carcinoma; RMS = root mean square; SEM = standard error of the mean; TTFields = Tumor Treating Fields.			cancers14122959_makarova	0
+Is that timing enough to get protein expression?			The manuscript is an interesting work related to a potential new therapy of hepatocellular carcinoma (HCC) by using TTFields in combination with a TKI (Sorafenib).			cancers14122959_makarova	0
+In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			Maybe the cytotoxicity data will reveal a different better frequency.			cancers14122959_makarova	0
+Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F.			cancers14122959_makarova	0
+*p < 0.05 relative to control; one-way ANOVA.			Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			cancers14122959_makarova	0
+When compared with sorafenib alone there is practically no difference.			Moreover, in the Discussion section, the authors concluded that “TTFields display efficacy for treatment of HCC in vitro and in vivo, with an optimal frequency of 150 kHz”.			cancers14122959_makarova	0
+However, the limited experimental design and paucity of strong data ask for more experiments to proof the feasibility of this combination for treating HCC.			*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			cancers14122959_makarova	0
+ANOVA = analysis of variance; HCC = hepatocellular carcinoma; RMS = root mean square; SEM = standard error of the mean; TTFields = Tumor Treating Fields.			GRP78 levels in the groups treated with TTFields or sorafenib alone remained unchanged from the control, but were elevated 2-fold in the TTFields plus sorafenib group (Figure 4f).			cancers14122959_makarova	0
+Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F.			Point 1: What is the rationale of using 150 kHz TTFields?			cancers14122959_makarova	0
+Overall, the authors believe that this research demonstrates potential for concomitant TTFields and sorafenib application in the treatment of HCC.			While I do not know if the authors have the technology to perform TTFields in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells.			cancers14122959_makarova	0
+Also, is there any evidence that TTF may prevent the pretty common resistance to sorafenib observed in clinic?			Since the authors missed to add Figure 4A for timeline, based on Methods section the rats were treated for 5 days with TTFields and or sorafenib and a day later the rats were sacrificed.			cancers14122959_makarova	0
+Response to Reviewer 1 Comments In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			Point 2: Despite the expression of cleaved PARP was very low in the tumors treated with TTFields or sorafenib alone (as shown in IHC-images in Figure 4F), the authors declare about ~ 20% of positive cells, as show in the graphs below IHC-staining.			cancers14122959_makarova	0
+Response 5: We thank the reviewer for this question.			In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			cancers14122959_makarova	0
+Point 4: Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib (when compared to the cells treated with TTFields and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 (for both HCC cell lines).			Several data have shown very high error bars in each group (especially Fig Can the author provide the tumor images which were harvested from mice?			cancers14122959_makarova	0
+Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.			Response to Reviewer 1 Comments In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			cancers14122959_makarova	0
+Response 6: We thank the reviewer for this comment.			I read the manuscript with interest and commend the authors for the work done in the area of liver cancer treatment.			cancers14122959_makarova	0
+Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group.			cancers14122959_makarova	0
+Author Response Please see attachment Author Response File: Author Response.docx			Why did the authors use high frequency of TTFields though the cytotoxicity was also observed in lower dose (Fig 1A).			cancers14122959_makarova	0
+There is a big difference between a therapy which kills vs a therapy which induces a cellular arrest.			Therefore, I expect further additional examination in the future.			cancers14122959_makarova	0
+I have raised the following concerns which is necessary to make this manuscript more scientifically interesting.			What are the p53 status and the apoptosis signaling pathway function in N1S1 cells?			cancers14122959_makarova	0
+This is an interesting scientific study, as it concerns the issue of hepatocellular carcinoma, it also has a clinical aspect.			TTFields frequency scan in rat N1S1 HCC cells.			cancers14122959_makarova	0
+The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			Author Response Please see attachment Author Response File: Author Response.docx			cancers14122959_makarova	0
+Response 7: We thank the reviewer for this question.			In the animal study we have now added IHC examination of beclin-1 and of GRP78, a marker for ER stress, as described in results sub-section 3.4.			cancers14122959_makarova	0
+In addition to IHC, I suggest performing western blot using PARP antibody where the full length and cleaved bands are observed in the same blot.			Point 6: Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.			cancers14122959_makarova	0
+I read the manuscript with interest and commend the authors for the work done in the area of liver cancer treatment.			Overall, the authors believe that this research demonstrates potential for concomitant TTFields and sorafenib application in the treatment of HCC.			cancers14122959_makarova	0
+GRP78 levels in the groups treated with TTFields or sorafenib alone remained unchanged from the control, but were elevated 2-fold in the TTFields plus sorafenib group (Figure 4f).			 Point 7: Why there is no error bar in the control group of all bar graphs?			cancers14122959_makarova	0
+In this manuscript, the authors, Davidi et al have investigated the effect of TTFields in HCC cells and an animal model, alone or in combination with sorafenib.			*p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 relative to time-respective control; two-way ANOVA.			cancers14122959_makarova	0
+HepG2 cells were treated for 6, 24, or 48 hours with 150 kHz TTFields, 3 µM sorafenib, or the two treatments combined, followed by Western blot examination of the autophagy markers beclin-1 and LC3 (d), the ER stress marker GRP78 (e), and the apoptosis marker cleaved PARP (f).			This might be helpful and make the in vitro data more relevant with the data shown in vivo.			cancers14122959_makarova	0
+Response 4: We thank the reviewer for this comment.			Response 7: We thank the reviewer for this question.			cancers14122959_makarova	0
+Point 1: In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells).			Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			cancers14122959_makarova	0
+As shown in the Figure 4 C and D, TTFields were found less effective in terms of reducing the tumors volume and weight when compared with sorafenib.			Were these western blots (Figure 3D-F) repeated 3 times as the material and method mentions?			cancers14122959_perova	0
+ Point 7: Why there is no error bar in the control group of all bar graphs?			Point 4: Since Annexin V/7-ADD data was not convincing and the authors observed the minor increase of apoptotic cells after HCC cells were treated with combination of TTFields and sorafenib (when compared to the cells treated with TTFields and sorafenib alone), I suggest to run the WBs to examine the expression of the cleaved forms of PARP and caspase-3 (for both HCC cell lines).			cancers14122959_perova	0
+While I do not know if the authors have the technology to perform TTFields in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells.			However, I feel that the experiments around this concept were not designed well enough to support their new findings.			cancers14122959_perova	0
+We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results.			Point 9: Was Mycoplasma testing done routinely?			cancers14122959_perova	0
+Similarly, low evidence of apoptosis (expression of cleaved PARP) was found in these groups, as shown in Figure 4F.			Overall, the authors believe that this research demonstrates potential for concomitant TTFields and sorafenib application in the treatment of HCC.			cancers14122959_perova	0
+The materials and methods section are elaborated in the details.			However, the exact mechanism of the combination therapy induced cell death is not yet known as the activation of autophagy in the combination therapy was not increased.			cancers14122959_perova	0
+These additions may be seen in Figure 3 and are described in results sub-section 3.4, Autophagy-apoptosis Interplay For Treatment with Concomitant TTFields and Sorafenib: “In order to investigate the mechanism of action of TTFields-sorafenib co-application, HepG2 and Huh-7D12 cells were treated for 6, 24, or 48 hours with TTFields, sorafenib (3µM), or the two modalities together, and then examined for expression levels of various proteins.			As a rule of thumb, when a housekeeping protein gives problems in immunoblotting, there are many other classic protein to switch the investigation (b-actin, vinculin, Ku, etc.).			cancers14122959_perova	0
+However, the limited experimental design and paucity of strong data ask for more experiments to proof the feasibility of this combination for treating HCC.			Is this indeed an anti-tumor effect or a very toxic effect to the liver (including tumor).			cancers14122959_perova	0
+While I do not know if the authors have the technology to perform TTFields in mice, where for sure they should have done xenograft models with the two human cell lines, why the rat cell line was not studied in vitro using the same experimental strategies as for human cells.			Also mention the ‘n’ number in all the experiments involved for invitro and invivo.			cancers14122959_perova	0
+Majority of rodent study in cancer research lose the control group early but the treated groups are followed up for much longer to delineate indeed the efficacy vs. toxicity.			The authors suggested that the therapeutic effects of the combination were apoptosis via autophagy.			cancers14122959_perova	0
+However, I feel that the experiments around this concept were not designed well enough to support their new findings.			The higher changes in expression levels and faster kinetics when TTFields and sorafenib were applied together rather than alone indicate higher stress levels imposed on the cells in the former case.” Figure 3.			cancers14122959_perova	0
+Similar, the graphs illustrating the LC3 expression are not in a proper fit with the images shown in Figure 4D.			Point 9: Was Mycoplasma testing done routinely?			cancers14122959_perova	0
+The in vivo model appears to be performed only time which question the validity of data.			Author Response Please see attachment Author Response File: Author Response.docx			cancers14122959_perova	0
+The apoptosis marker cleaved PARP displayed increased expression in the combined group already after 24 hours, elevating even further after 48 hours of treatment.			Point 3: It will be much better to provide the data to explain the mechanisms illustrating why the monotherapy of TTFields or sorafenib induced autophagy, whereas the tumors treated with combination developed the substantial apoptotic death of tumor cells.			cancers14122959_perova	0
+The rigor science recommends at least 2 independent experiments with at least 7 animals randomized per group.			At least one different dose should have been studied for comparison since this is a completely different tumor environment than the in vitro one.			cancers14122959_perova	0
+We thank the reviewer for this comment, as these additions add much clarity to the mechanism of action of TTFields in combination with sorafenib and provide a more coherent explanation for the in vivo results.			Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is totally different?			cancers14122959_perova	0
+The authors indicated that TTField had potential to be a new treatment option of hepatocellular carcinoma.			I have raised the following concerns which is necessary to make this manuscript more scientifically interesting.			cancers14122959_perova	0
+For the monotherapies, cleaved PARP increase was only evident at 48 hours of treatment, and to a lower extent than that in the co-treatment group (Figure 3f).			Response: We thank the reviewer for the positive review.			cancers14122959_perova	0
+Were these western blots (Figure 3D-F) repeated 3 times as the material and method mentions?			Can this dose observed in vitro on only tumor cells be translated to in vivo work where the tumor microenvironment is totally different?			cancers14122959_perova	0
+Were the mice perfused before collecting the tumors?			Point 8: Since sorafenib acts also on angiogenesis, did the authors investigate if TTFields may interfere with anti-angiogenic effect sorafenib-mediated?			cancers14122959_perova	0
+Overall, the authors believe that this research demonstrates potential for concomitant TTFields and sorafenib application in the treatment of HCC.			Point 6: Any explanation for why not using cloroquine in vivo to integrate better the in vitro data.			cancers14122959_perova	0
+The reference protein should have the same intensity since this is the control for equal protein loading.			The rigor science recommends at least 2 independent experiments with at least 7 animals randomized per group.			cancers14122959_perova	0
+This might be achieved by using the combination of so-called Tumor Treating fields (TTFields) with targeted drug, I have the following suggestions about this manuscript: Point 1: The authors demonstrate the efficacy of TTFields in vivo even when used as monotherapy.			Point 1: In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells).			cancers14122959_perova	0
+Were the cells from supernatant counted (where are probably the majority of dead cells)?			Response 6: We thank the reviewer for this comment.			cancers14122959_perova	0
+Moreover, there is a discrepancy between the fold changes in tumor weight vs. volume in the combination group vs. untreated group.			I read the manuscript with interest and commend the authors for the work done in the area of liver cancer treatment.			cancers14122959_perova	0
+Point 1: In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells).			Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.			cancers14122959_perova	0
+Point 1: In vitro experiments sub-section in Methods sections lacks many experimental details (e.g., type of plate/flask, plating overnight or not before experiment, number of plated cells).			Concomitant TTFields with Sorafenib Enhances Treatment Efficacy in Vivo: “Tumor histology and immunostaining for beclin-1 and LC3, GRP78, and cleaved PARP were performed to examine autophagy, ER stress, and apoptosis levels, respectively.			cancers14122959_perova	0
+Author Response Please see attachment Author Response File: Author Response.docx			Is that timing enough to get protein expression?			cancers14122959_perova	0
+Moreover, many bands belonging to all investigated proteins are truncated, fractured and I identified a lot of troubleshooting in bands due the presence of bubbling when running the blots.			The autophagy marker LC3 also displayed such bi-phasic characteristics, but with a somewhat slower kinetics, showing some elevation at 6 hours of treatment, but higher elevation at the 24 hours time point (Figure 3d).			cancers14122959_perova	0
+The assay results enabled direct measurement of the percentage of the isoforms generated thus facilitating a better understanding of the dominant isoform generated by each variant and their effect on protein coding.			As such, in order to correlate the discussion in this paragraph with the information in table 2, the reader also needs to cross reference Table 1 or 3.			cancers14122960_makarova	0
+I would also stress a little more that FL is almost undetectable.			Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively.			cancers14122960_makarova	0
+The authors further used the minigene splicing assay results, and understanding of RAD51C protein biology, to evaluate each variant using the most current ACMG-AMP frameworks for variant classification.			Author Response Thank you very much for the positive comments 1-			cancers14122960_makarova	0
+Is it known whether there is a threshold of RAD51C deficiency that is tolerated before associated cancer risks become increased?			"The manuscript entitled ""Minigene splicing assays identify 20 spliceogenic variants of the breast/ovarian cancer susceptibility gene RAD51C authored by Sanoguera-Miralles et al., is a very interesting study."			cancers14122960_makarova	0
+However, the corresponding tables that summarizes variant classification according to the ACMG/AMP-based criteria (Table 2), does not include these transcript isoform names.			Although a reference is provided so that the reader can look up what these ad-hoc rules are, it would also be helpful to briefly describe these in the current manuscript.			cancers14122960_makarova	0
+Leaking splicing was also noted in two instances resulting in detection of the canonical transcript in 26.3% and 21.3% of transcripts, respectively.			Although a reference is provided so that the reader can look up what these ad-hoc rules are, it would also be helpful to briefly describe these in the current manuscript.			cancers14122960_makarova	0
+Is there evidence from another source that these beta strands are critical to protein function and that their loss is deleterious (rather than resulting in normal or slightly reduced protein activity)?			The transcript analysis results demonstrated splice complexity with several variants resulting in two or more transcript isoforms.			cancers14122960_makarova	0
+While in silico prediction tools can provide insights to possible impacts of nucleotide variants on splicing, these prediction tools do not replace the empirical evidence that the minigene assay can provide.			These variant interpretations were generally evidence based and well thought out.			cancers14122960_makarova	0
+Overall manuscript is well written, very relevant for variant analysis and is suitable for publication if they address the following concerns: 1)			The transcript analysis results demonstrated splice complexity with several variants resulting in two or more transcript isoforms.			cancers14122960_makarova	0
+present an interesting manuscript that utilizes a minigene splicing assay to examine twenty RAD51C variants catalogued through ClinVar.			The manuscript is very well written, clear and well discussed; I have just few comments: in the Introduction in the sentence 61-69 the authors do not mention anything about BRCA1 (they mention MLH1 though); I would expect that as BRCA1 splice variants are deeply studied.			cancers14122960_makarova	0
+The manuscript is very well written, clear and well discussed; I have just few comments: in the Introduction in the sentence 61-69 the authors do not mention anything about BRCA1 (they mention MLH1 though); I would expect that as BRCA1 splice variants are deeply studied.			Lines 342-359 – This paragraph discusses two RAD51C variants, c.404+3A>G and c.705+3A>G, for which mg-FL transcripts were detected in 26.3% and 21.3% or transcripts, respectively.			cancers14122960_makarova	0
+Results section (Page 6, line 221): We investigated the association of NR2F1 expression with metastasis.			Cell proliferation-related gene sets were suppressed and MKi67 expression was lower in high NR2F1 BC.			cancers14122962_makarova	0
+Discussion section (Page 13, line 427): The current study demonstrated that NR2F1 expression in the bulk tumor of primary breast cancer is associated with decreased cell proliferation and cancer stem cell-like characteristics.			In addition, it shows that NR2F1 is primarily expressed in CAFs, particularly in inflammatory CAFs.			cancers14122962_makarova	0
+Conclusions: NR2F1 expression in breast cancer is associated with tumor dormancy traits and it is predominantly ex-pressed in CAFs in the tumor microenvironment.			Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs.			cancers14122962_makarova	0
+Can the authors do some analyses using data generated from both primary and metastatic tumors to check if there is any difference of NR2F1 expression, and how NR2F1 expression is correlated with metastasis?			Expression of NR2F1 between endocrine therapy responder and non-responder in hormone-positive primary breast cancer.			cancers14122962_makarova	0
+Kaplan-Meier curves of overall survival (OS), disease- specific survival (DSS), and disease-free survival (DFS) based on the high and low NR2F1 expression in triple-negative breast cancer of three large cohorts.			(A) Boxplots showing various scores based on high and low NR2F1 expression in triple-negative breast cancer of TCGA; Intratumoral heterogeneity, homologous recombination deficiency (HRD), silent/non- silent mutation rate, SNV/Indel neoantigen, Interferon gamma response, fraction altered, tumor-infiltrating lymphocytes (TIL) fraction, and stromal fraction.			cancers14122962_makarova	0
+Kaplan-Meier curves of overall survival (OS), disease- specific survival (DSS), and disease-free survival (DFS) based on the high and low NR2F1 expression in triple-negative breast cancer of three large cohorts.			Comment 2: The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels.			cancers14122962_makarova	0
+Previous studies mentioned that NR2F1 was highly expressed in DTCs [67].			Comment 3: Line 170, “Figure 1E” should be “Figure 1D”.			cancers14122962_makarova	0
+In order to prove that CAF-expressed NR2F1 regulates breast tumor dormancy, one needs to analyze the single-cell sequencing data and show whether the late recurrence in other organs, such as lungs, bones, and brain, other than lymph nodes, is correlated with NR2F1 expression in the CAFs.			We found that NR2F1 is most predominantly expressed in CAFs in the TME of primary breast cancer.			cancers14122962_makarova	0
+Comment 2: The authors choose chemotherapy as a treatment option to compare it with NR2F1 levels.			Methods: A total of 6758 transcriptomes of bulk tumors from multiple breast cancer patient cohorts and two single-cell sequence cohorts were analyzed.			cancers14122962_makarova	0
+Results section (Page 11, line 346): There was no difference in NR2F1 expression between responder and non-responder to neoadjuvant endocrine therapy (Supplementary Fig. 9).			Thus, we investigated the clinical relevance of the expression of NR2F1, a known dormancy biomarker.			cancers14122962_perova	0
+However, the findings from our study are not ample to substantiate that CAF-expressed NR2F1 regulates breast tumor dormancy.			Please find our point-by-point responses below.			cancers14122962_perova	0
+Association of NR2F1 with immunity within the tumor microenvironment of HER2 positive breast cancer.			We found that NR2F1 is most predominantly expressed in CAFs in the TME of primary breast cancer.			cancers14122962_perova	0
+Discussion section (Page 13, line 427): The current study demonstrated that NR2F1 expression in the bulk tumor of primary breast cancer is associated with decreased cell proliferation and cancer stem cell-like characteristics.			Man-Whitney U test was used to compare the two groups and p-values are shown in bold for significant results (p < 0.05).			cancers14122962_perova	0
+Also, dot plots show the expression of NR2F1, TGFB1, SOX9, and RARB by cell type in each immunohistological subtype in single-cell Cohort 2.			To this end, we believe that NR2F1 expression in the bulk tumor does not reflect the expression in the cancer cells, thus its value as a biomarker is in doubt.			cancers14122962_perova	0
+Results section (Page 11, line 346): There was no difference in NR2F1 expression between responder and non-responder to neoadjuvant endocrine therapy (Supplementary Fig. 9).			Kaplan-Meier curves of overall survival (OS), disease- specific survival (DSS), and disease-free survival (DFS) based on the high and low NR2F1 expression in triple-negative breast cancer of three large cohorts.			cancers14122962_perova	0
+Log-rank test was used for the analysis, and significant p values are shown in bold.			Nat Cell Biol 2017), it is puzzling how NR2F1 in CAFs of the primary tumor would contribute to dormancy of DTCs.			cancers14122962_perova	0
+Comment 2: The authors should clearly explain the innovation and importance of their work on the introduction of the manuscript.			They should justify the value of the work and compare their work with previously similar published papers.			catal12030290_makarova	0
+According to the reviewer suggestion, the SEM images with same scale are provided in the revised manuscript.			Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+Comment 5: Why authors did not use XRD technique for sample characterization?			A point-by-point response to the reviewer-2 comments is appended below for your convenience.			catal12030290_makarova	0
+Synthesis of bioadsorbent and evaluation of its structure using SEM and FT-IR spectroscopy.			Comment 2: The authors should clearly explain the innovation and importance of their work on the introduction of the manuscript.			catal12030290_makarova	0
+: catalysts-1584262 Title: Bio-stimulated adsorption of Cr(VI) from aqueous solution by Groundnut Shell Activated Carbon@Al embedded material Response to Reviewer-2 Comments We appreciate the efforts of the reviewers for their detailed and insightful comments, which have helped us to improve the quality of our manuscript.			All the modifications are shown in yellow color in the revised manuscript.			catal12030290_makarova	0
+We hope the reviewer understand the experimental deficiencies at the stage of the present experiments.			There are some points which must be edited or clarified by providing additional information or comments:			catal12030290_makarova	0
+Comments to the Author Manuscript Catalysts-1584262 The manuscript entitled ‘Bio-stimulated adsorption of Cr(VI) from aqueous solution by Groundnut Shell Activated [email protected] embedded material’ by Rao et al focuses on the synthesis of bioadsorbent aluminum metal blended with groundnut shell activated carbon material (Al-GNSC) and it practical application for Cr(VI) adsorption from waste aqueous solutions.			Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+(ppm) pH Contact Time (min) Adsorbent dosage (g l-1) Maximum adsorption capability (mg g-1) References Activated carbon (AC) prepared from coconut tree sawdust 10 3.0 180 0.2 3.46 [29] Raw coconut fiber 250 1.0 270 10 18.60 [30] Sugarcane bagasse 100 2.0 90 10 1.76 [31] Canadian peat Coconut fiber 50 2.0 4320 25 4.61 4.71 [32] peanut shell (P. Shell), sawdust (S. Dust) and Cassia fistula leaves (C.F.			Comment 2: The porosity of the sample should be measured.			catal12030290_makarova	0
+There are some points which must be edited or clarified by providing additional information or comments:			After making all required minor changes in article it could be recommended for publication.			catal12030290_makarova	0
+Please add the discussion which peak shift is related to Cr, the authors can combine these with the discussion of the mechanism.			The authors are very thankful to the Reviewer for their valuable suggestions for the improvement of the manuscript.			catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			First of all, authors have to attach a EDX mapping images before/after sorption of chromium ions.			catal12030290_makarova	0
+Response: We acknowledge the reviewer’s opinion.			The authors are very thankful to the Reviewer for their valuable suggestions for the improvement of the manuscript.			catal12030290_makarova	0
+We hope the reviewer understand the experimental deficiencies at the stage of the present experiments.			Page 2 of 2 Comment 5: In the experiment, H2SO4 is added in the first step, what is the purpose?			catal12030290_makarova	0
+Comment 9: The conclusion section should be elaborated and improved.			 Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+Comment 8: In order to confirm proposed mechanism of Cr(VI) adsorption (illustrated on the fig 6) Authors should provide data on adsorption capacity of pristine groundnut shell activated carbon (not modified with Al).			Response: We acknowledge the reviewer’s opinion.			catal12030290_makarova	0
+Examination of prepared adsorbents for Cr(VI) ions removal.			However, we regret that we were not able to investigate the BET analysis due to pandemic situation, which could definitely give us additional information.			catal12030290_makarova	0
+The authors are very thankful to the Reviewer for their valuable suggestions for the improvement of the manuscript.			Is the products suitable for this real condition?			catal12030290_makarova	0
+In such form is rather difficult to make adequate comparison.			Please add the discussion which peak shift is related to Cr, the authors can combine these with the discussion of the mechanism.			catal12030290_makarova	0
+Furthermore, it should be justified why this difference does not influence the comparison of the results with the different cell lines (THP1, Jurkat, Raji).			– the activity of PLY/SLO was prevalent at the initial times of incubation and at relatively high amounts (volume) of supernatants, whereas SLS activity fully developed only after initial lag period but was prevalent when relatively low amounts of supernatants were used in the assays.			cells11010166_perova	0
+Point-by-point response to reviewers: Reviewer 1 comments 1.			Its exact mode-of-action is still not yet fully clarified.			cells11010166_perova	0
+However, given the poor characterization of SLS, its unavailability from commercial providers and its extremely tedious purification, the mechanistic details of the SLS neutralization will require an extensive a project of its own and are beyond the scope of the current study.			This control represents 100% survival and allows us to normalize our survival data.			cells11010166_perova	0
+In one control we challenged the cells with bacterial supernatant without adding liposomes, to determine that the baseline cytotoxicity is in line with the results displayed in the Figure 1.			Figure 2 (for referee inspection only) demonstrates that Alamar blue cell viability assay and Trypan blue live/dead quantification provide results that are identical to those obtained in the cell proliferation protocol used in our study.			cells11010166_perova	0
+The following text was added to the material and method section: The data were normalized to a control incubated with PBS instead of bacterial supernatant (considered as 0% cell death).			We hope that with these changes our manuscript will be considered for publication and we are looking forward to hearing from you.			cells11010166_perova	0
+However, as the immune cell lines have different sensitivities depending on the toxin profile of the tested strain, a similar lethal dose corresponds to different supernatant volumes.			However, authors did not make any attempt to explore this aspect in mechanistic detail.			cells11010166_perova	0
+Figure 2 (for referee inspection only) demonstrates that Alamar blue cell viability assay and Trypan blue live/dead quantification provide results that are identical to those obtained in the cell proliferation protocol used in our study.			It is not clear why they are calling the preparations as the nanotraps.			cells11010166_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_makarova	0
+Second Round of Evaluation Round 2: and Author Response			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+Second Round of Evaluation Round 2: and Author Response			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_makarova	0
+The article is well written, and the overall study was well planned and professionally conducted.			However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_makarova	0
+Second Round of Evaluation Round 2: and Author Response			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_makarova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+Second Round of Evaluation Round 2: and Author Response			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+All the experiments shown are necessary and sufficient to support the conclusions.			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+The article is well written, and the overall study was well planned and professionally conducted.			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			All the experiments shown are necessary and sufficient to support the conclusions.			cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			However, I think that the article can be substantially improved if the following points are addressed: (1)			cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+However, I think that the article can be substantially improved if the following points are addressed: (1)			Second Round of Evaluation Round 2: and Author Response			cosmetics2030248_perova	0
+Round 1: and Author Response The article titled “Thermal behavior and free-radical-scavenging activity of phytic acid alone and incorporated in galenic emulsions” is focused on the characterization of phytic acid stability and use in cosmetic formulas.			The article is well written, and the overall study was well planned and professionally conducted.			cosmetics2030248_perova	0
+This makes the unconditional correlations uninteresting.			References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			economies3040150_perova	0
+More generally, we venture the highly subjective view that despite the immense mathematical sophistication of cointegration tests and their application in perhaps thousands of studies, it is not obvious how much additional substantive insight has been gained in the wide variety of contexts in which these tests have been used.”			Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			economies3040150_perova	0
+The suggestions are listed below in no particular order of importance: 1.			This makes the unconditional correlations uninteresting.			economies3040150_perova	0
+Penn World Table Revisions and Their Impact on Growth Estimates” NBER Working Paper No.			Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+Round 1: The paper analyzes the validity of Wagner’s law in six East Asian countries during the period 1960- 2008.			Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			economies3040150_perova	0
+Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			economies3040150_perova	0
+Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			This makes the unconditional correlations uninteresting.			economies3040150_perova	0
+More generally, we venture the highly subjective view that despite the immense mathematical sophistication of cointegration tests and their application in perhaps thousands of studies, it is not obvious how much additional substantive insight has been gained in the wide variety of contexts in which these tests have been used.”			While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			economies3040150_perova	0
+Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			economies3040150_perova	0
+Penn World Table Revisions and Their Impact on Growth Estimates” NBER Working Paper No.			Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+This makes the unconditional correlations uninteresting.			Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			economies3040150_perova	0
+Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			economies3040150_perova	0
+At any rate, it is not obvious that such sophisticated and complex tests of cointegration provide any useful additional insight regarding the empirical status of the hypothesis.			Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			economies3040150_perova	0
+Round 1: The paper analyzes the validity of Wagner’s law in six East Asian countries during the period 1960- 2008.			Round 2: Author Response to Reviewer 2 Thank you.			economies3040150_perova	0
+While interesting, generally well written and scientifically sound, several suggestions should be considered before further consideration.			Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			economies3040150_perova	0
+Second Round of Evaluation Round 2: The paper is much better and I only have some minor comments.			References: Cameron A. Shelton (2007) “The size and composition of government expenditure” Journal of Public Economics, 91 (11–12) Simon Johnson, William Larson, Chris Papageorgiou, Arvind Subramanian.			economies3040150_perova	0
+Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			Moreover, most studies that add variables to the models, such as the dependency ratio, argue that Wagner’s law holds conditional on it.			economies3040150_perova	0
+General comments The authors apply Latent Growth Curve Model (LGCM) approach to characterize the growth path of China’s agricultural labor productivity between 2000 and 2010 using a balanced panel data.			Maybe, the footnote 5 could go to the table 2’s note.			economies4030013_makarova	0
+Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.			Maybe, the footnote 5 could go to the table 2’s note.			economies4030013_makarova	0
+In general, we amended the paper in accordance with the replies below.			This could have strong implications in one case or another, especially in the first case.			economies4030013_makarova	0
+- We would like to thank the reviewer’s scrutiny.			This could have strong implications in one case or another, especially in the first case.			economies4030013_makarova	0
+9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.			One of the most interesting issues of the paper is the political change in 2004.			economies4030013_makarova	0
+For this reason we use the provincial price indices to deflate the nominal series.			We summarize below our reactions to the points raised by the Reviewers following the same order in the lists of comments.			economies4030013_makarova	0
+So, if the agricultural output is in current prices, the growing prices will affect deeply the results.			We now include more recent literature in the introduction, as well as the government movements of the relevant policies in recent years.			economies4030013_makarova	0
+Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.			Issues to improve the paper: The introduction is welldone, but it could be improved in some points.			economies4030013_makarova	0
+Hence, all the prefectures within each province are deflated by the same index.			I think, this clear objective could improve the introduction about the target that the authors follow in this paper.			economies4030013_makarova	0
+Maybe, the footnote 5 could go to the table 2’s note.			Although suboptimal, this helps to preserve the comparability in the two estimations.			economies4030013_makarova	0
+Author Response We would like to thank the Reviewers for a very careful reading of the paper and a number of interesting, though challenging, suggestions and comments.			The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			economies4030013_makarova	0
+Are the annual agricultural output data are measured in their nominal value (at current price) or in real value (at constant price)?			General comments The authors apply Latent Growth Curve Model (LGCM) approach to characterize the growth path of China’s agricultural labor productivity between 2000 and 2010 using a balanced panel data.			economies4030013_makarova	0
+Appendix Figure A2: The figure presents the chain-linked index of output and input.			Besides, the authors must define better the data how the agricultural labor productivity was calculated.			economies4030013_perova	0
+General comments The authors apply Latent Growth Curve Model (LGCM) approach to characterize the growth path of China’s agricultural labor productivity between 2000 and 2010 using a balanced panel data.			For this reason we use the provincial price indices to deflate the nominal series.			economies4030013_perova	0
+Appendix Figure A2: The figure presents the chain-linked index of output and input.			Besides, the implications of this political reform are explained in the text, but more detail in these explanations would permit to understand better these changes.			economies4030013_perova	0
+The agricultural output is in current or constant prices?			The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			economies4030013_perova	0
+9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.			While the topic is interesting there are some major issues that need to be properly addressed first.			economies4030013_perova	0
+There is no price index at the prefectural level.			We would like to thank the Reviewer to recommend us to develop this part.			economies4030013_perova	0
+As a suggestion for this paper or another, having the data of output, labor and land, it is also interesting to analyze the agricultural labor productivity explained by the agricultural land productivity and land-labor ratios.			Appendix Figure A2: The figure presents the chain-linked index of output and input.			economies4030013_perova	0
+General comments The authors apply Latent Growth Curve Model (LGCM) approach to characterize the growth path of China’s agricultural labor productivity between 2000 and 2010 using a balanced panel data.			It is not clear whether the output is measured in real value (at constant price) or nominal value (at current price).			economies4030013_perova	0
+If this “first sector” includes the fish workers, the agricultural labor productivity measures are not correct in the regions with sea access.			The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			economies4030013_perova	0
+The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			We would like to thank the Reviewer to recommend us to develop this part.			economies4030013_perova	0
+If it’s based on the real value then what kind of deflators are used in the estimation, national level or regional level since agricultural prices may vary across regions?			The authors could enrich the results’ part with reference to the concepts of convergence of Barro and Sala-i-Martin, beta and sigma convergence.			economies4030013_perova	0
+9 lines 5-6, but they can point out the non existence in the case of Chinese prefectural level.			The context of this paper consists in growing agricultural prices, especially in the second half of this decade in the world panorama.			economies4030013_perova	0
+Comment) Conclusions: Future directions should be added to the Conclusion Section.			Some references do not contain enough information such as [16], [18] … etc.			electronics11050760_makarova	0
+The list of references requires an extensive check.			Fast DST-VII/DCT-VIII With Dual Implementation Support for Versatile Video Coding, In IEEE Transactions on Circuits and Systems for Video Technology, 2021, vol.			electronics11050760_makarova	0
+The revised manuscript was reviewed and substantially modified by a native speaker.			According to a reviewer’s comment, we included two new Tables, Table 3 and Table 4, so that the table indices were changed accordingly.			electronics11050760_makarova	0
+Dong, X.; Shen, L.; Yu, M.; Yang, H. Fast Intra Mode Decision Algorithm for Versatile Video Coding.			The results show a higher performance of the proposed computation of inverse transforms.” We would like to thank the reviewer for the thorough review and very helpful comments.			electronics11050760_makarova	0
+Ref [31] was removed because the paper was nothing to do with the proposed manuscript.			IEEE Transactions on Circuits and Systems for Video Technology, 2020, vol.			electronics11050760_makarova	0
+Inverse Transform Using Linearity Hyeonju Song*, and Yung-Lyul Lee* *Digital Media System Laboratory, Department of Computer Engineering, Sejong University, Seoul, Korea.			Han, J.; Saxena, A.; Melkote, V.; Rose, K. Jointly Optimized Spatial Prediction and Block Transform for Video and Image Coding, In IEEE Transactions on Image Processing, 2012, vol.			electronics11050760_makarova	0
+The authors should add the main contributions briefly at the end of the introduction.			It is better that they replace them with other keywords to increase the reach of the manuscript.			electronics11050760_makarova	0
+The authors should accurately address the below comments.			Koo, M.; Salehifar, M.; Lim, J.; Kim, S. -H. CE6-Related: 32 Point MTS Based on Skipping High Frequency Coefficients.			electronics11050760_makarova	0
+Also, they should clarify the limitations of the proposed method.			In Proceedings of the 13th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Marrakech, Morocco, 9-18 January 2019; 17.			electronics11050760_makarova	0
+We included the following sentence and Equation (14) on page 4: “For an (n×m) transform block which has N non-zero coefficients, the total number of additions in the proposed inverse transform using linearity is computed in Equation (14), even though Equation (14) is not used in the proposed method: We included the following Table 3 and 4 and sentences on page 6: Table 3.			We would like to thank the reviewer for the thorough review and very helpful comments.			electronics11050760_makarova	0
+In Proceedings of the 14th Meeting Joint Video Experts Team (JVET) of ITU-T SG 16 WP 3 and ISO/IEC JTC 1/SC 29/WG 11, Geneva, Swiss, 19-27 March 2019; 19.			Comment) Conclusions: Future directions should be added to the Conclusion Section.			electronics11050760_makarova	0
+The results show a higher performance of the proposed computation of inverse transforms.			Also, they should clarify the limitations of the proposed method.			electronics11050760_makarova	0
+IEEE Access, 2019, vol. 7, pp. 158075-158084, doi: 10.1109/ACCESS.2019.2950388.			• Discussion Section: The authors did not respond accurately to this comment.			electronics11050760_perova	0
+However, the proposed method can be considered in the next-generation video coding standards because it achieves decoding run-time saving, while maintaining average BD-rate.			Author Response Please find the attached file.			electronics11050760_perova	0
+The list of references requires an extensive check Answer)			Also, they should clarify the limitations of the proposed method.			electronics11050760_perova	0
+Koo, M.; Salehifar, M.; Lim, J.; Kim, S. -H. CE6-Related: 32 Point MTS Based on Skipping High Frequency Coefficients.			Answer) According to the reviewer’s comment, the revised manuscript was reviewed and substantially modified by an English native speaker.			electronics11050760_perova	0
+Also, they should clarify the limitations of the proposed method.			Also, they should clarify the limitations of the proposed method.			electronics11050760_perova	0
+We included the following sentence and Equation (14) on page 4: “For an (n×m) transform block which has N non-zero coefficients, the total number of additions in the proposed inverse transform using linearity is computed in Equation (14), even though Equation (14) is not used in the proposed method: We included the following Table 3 and 4 and sentences on page 6: Table 3.			Some links do not work in the reference list like [22] … etc.			electronics11050760_perova	0
+and others use only an uppercase letter in the first word (such as [2], [9] … etc.			Bjontegaard, G. Calculation of Average PSNR Differences Between Rd-Curves, Austin, TX, USA, pp.			electronics11050760_perova	0
+As this discussion and comparison can clarify the fairness and rationality of the results of the proposed method.			Inverse Transform Using Linearity Hyeonju Song*, and Yung-Lyul Lee* *Digital Media System Laboratory, Department of Computer Engineering, Sejong University, Seoul, Korea.			electronics11050760_perova	0
+In answer to the specific comments: Comment 1) Introduction Section: This comment still requires a response.			Available online: https://vcgit.hhi.fraunhofer.de/jvet/VVCSoftware_VTM/-/tree/VTM-8.2 (accessed on 6 September 2021) 23.			electronics11050760_perova	0
+• Figures and Tables: All figures and tables are shown before they are used in the text.			Please add information on the number of additions for different transform types and sizes in original and proposed variants.			electronics11050760_perova	0
+Simulation results are presented to show the effectiveness of the proposed method.			Unless the quality of figures is not improved.			en15031006_perova	0
+Point 5: It could be better to include base load ratings in the results section (Fig: 10) Response 5:			Diesel generators equal to the rating of base load is kept as backup to avoid total black out in worse conditions.			en15031006_perova	0
+K-mean clustering is used to make clusters of the data based on two factors: probability of turning ON a specific appliance at time t and priority number given by consumer to that specific appliance.			Point 4: A comprehensive deeper literature review is necessary to address the research issue, Also authors need to provide a literature survey in an organized way.			en15031006_perova	0
+Response 4: According to your suggestion, we have modified the literature review and included some latest references i.e., [22][29][30][32].			K-mean clustering is used to make clusters of the data based on two factors: probability of turning ON a specific appliance at time t and priority number given by consumer to that specific appliance.			en15031006_perova	0
+Point 6: Sizing of renewable energy sources and battery systems were not properly explored.			Point 4: A comprehensive deeper literature review is necessary to address the research issue, Also authors need to provide a literature survey in an organized way.			en15031006_perova	0
+Point 3: The quality of the figures needs to be improved.			Diesel generators equal to the rating of base load is kept as backup to avoid total black out in worse conditions.			en15031006_perova	0
+As the proposed system is in isolated mode, how continuity of power supply be maintained?			Point 5: All the figures need to be improved.			en15031006_perova	0
+As the proposed system is in isolated mode, how continuity of power supply be maintained?			Point 4: A comprehensive deeper literature review is necessary to address the research issue, Also authors need to provide a literature survey in an organized way.			en15031006_perova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			Battery storage bank is included in the system to provide the baseload power in the time when renewable sources are not suffice to meet the baseload power demand.			en15031006_perova	0
+Point 5: All the figures need to be improved.			Point 2: In such a study an experimental case must be presented in a specified region Response 2: Thank you for your comment.			en15031006_perova	0
+The main contribution of this paper include: A demand side management algorithm is proposed to fulfil the energy gap between generation and consumer’s demand for standalone renewable energy system.			The appliances are prioritized based on their need and demand to achieve optimum energy utilization.			en15031006_perova	0
+"""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH."			Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.			fermentation8050212_makarova	0
+Standardize, taking into account the rules of the journal.			": fermentation-1703121 Title: Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation Overview and general recommendation: The article entitled ""Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation"", and submitted to publication in ""Fermentation"", is within the scope of the journal and discusses an important topic."			fermentation8050212_makarova	0
+1703121)’ for publication in the Fermentation.			It appears on Page 4, Line 162; Explain the role of sodium humate.			fermentation8050212_makarova	0
+1703121)’ for publication in the Fermentation.			It consists of performing a solid-state rice bran fermentation using a bacteria mix.			fermentation8050212_makarova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			I ask that the authors specifically address each of my comments in their responses.			fermentation8050212_makarova	0
+Major comments Sometimes the document is different to follow due to English issues.			Page 6, Lines 217-218: Check the values presented in the following sentence: “As shown in Table 1, the water-soluble polysaccharide content of FRBE and RBE were 552.30±2.68 mg g-1 and 450.28±1.69 mg g-1 respectively.”, because they are opposite to the ones presented in Table 1.			fermentation8050212_makarova	0
+For your suggestion we have dealt with the comments as follows: Comments to the Author			Four experimental groups were added AAPH and 0.1, 0.2, 0.3, 0.4 mg/ mL sample, respectively.”; Page 5, Lines 188-190: “The optimum sodium humate content was found to be 1.0 %, which DPPH free radical scavenging rate of 81.22%.”; Page 6, Lines 218-219: “The significantly less polysaccharide content of FRBE than that of RBE (P＜05).”; Page 8, Lines 241-242: “Schmidt et al.			fermentation8050212_makarova	0
+It appears for the first time on Page 1, Line 17; The hot water extraction conditions were tested before?			It appears for the first time on Page 1, Line 17; The hot water extraction conditions were tested before?			fermentation8050212_makarova	0
+Author Response Honored reviewers, Thank you for your contribution to reviewing work.			Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_makarova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			It consists of performing a solid-state rice bran fermentation using a bacteria mix.			fermentation8050212_makarova	0
+"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			"Page 4, Line 133: Change ""VWD detector"" to ""Variable wavelength detector (VWD)""."			fermentation8050212_makarova	0
+Author Response Honored reviewers, Thank you for your contribution to reviewing work.			"Page 3, Line 124: Change ""ethyl acetate reagents"" to "" ethyl acetate reagent""."			fermentation8050212_makarova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			fermentation8050212_makarova	0
+In the reference list, write the names of the microorganisms in italic.			Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			fermentation8050212_makarova	0
+"Page 3, Line 124: Change ""ethyl acetate reagents"" to "" ethyl acetate reagent""."			1703121)’ for publication in the Fermentation.			fermentation8050212_makarova	0
+Since I have some doubts that I would like clarified, I recommend that a major revision be warranted.			For your suggestion we have dealt with the comments as follows: Comments to the Author			fermentation8050212_makarova	0
+Why did the water-soluble phenolic acids contents increase after the fermentation?			1703121)’ for publication in the Fermentation.			fermentation8050212_makarova	0
+"Examples: Page 1, Lines 9-11: ""In order to enhancement the antioxidant properties of rice bran, solid state fermentation of rice bran with mixed bacteria was adopted and optimized fermentation technology."			Study on the effect of sodium humate on solid fermentation of lactobacillus plantarum.			fermentation8050212_perova	0
+The application of the Duncan test should be justified because it is less sensitive in this kind of study.			Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_perova	0
+Study on the effect of sodium humate on solid fermentation of lactobacillus plantarum.			": fermentation-1703121 Title: Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation Overview and general recommendation: The article entitled ""Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation"", and submitted to publication in ""Fermentation"", is within the scope of the journal and discusses an important topic."			fermentation8050212_perova	0
+For your suggestion we have dealt with the comments as follows:			Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_perova	0
+Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			For your suggestion we have dealt with the comments as follows: Comments to the Author			fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			For your suggestion we have dealt with the comments as follows: Comments to the Author			fermentation8050212_perova	0
+Table 1: The number of decimal places of some constituents is excessive.			(2014) also found that ferulic acid content of rice bran water-soluble extracts significant improvements, up to 765 mg/g [16].”; Page 10, Lines 321-323: “At the concentration of 0.3 mg/mL and 0.4 mg/mL, CAT activity induced by AAPH of FRBE group significantly higher than that of AAPH and RBE.”.			fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			Table 1: The number of decimal places of some constituents is excessive.			fermentation8050212_perova	0
+This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			fermentation8050212_perova	0
+"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			fermentation8050212_perova	0
+The article is a fair enough approach, please include a more specific section about the statistical methods.			Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_perova	0
+Since I have some doubts that I would like clarified, I recommend that a major revision be warranted.			"Minor comments Page 1, Line 9: Change ""tastea"" to ""taste""."			fermentation8050212_perova	0
+1703121)’ for publication in the Fermentation.			The appropriate methodology considering parametric assumption for normality and homogeneity of variances before data analysis should be applied.			fermentation8050212_perova	0
+For your suggestion we have dealt with the comments as follows:			Study on the effect of sodium humate on solid fermentation of lactobacillus plantarum.			fermentation8050212_perova	0
+Page 10, Lines 306 and 307: check the 73.85 and 77.00 values.			Author Response Honored reviewers, Thank you for your contribution to reviewing work.			fermentation8050212_perova	0
+Page 2, Lines 66 and 67: Write the microorganisms' names in italic.			Table 1: The number of decimal places of some constituents is excessive.			fermentation8050212_perova	0
+Table 1: The number of decimal places of some constituents is excessive.			Table 1: The number of decimal places of some constituents is excessive.			fermentation8050212_perova	0
+"""; Page 4, Lines 141-143: “In the morning (on set light), breeding 9 fishes interbreed 1h, embryos were obtained from natural spawning collection of embryos were completes in petri Dishes; Page 4, Lines 153-155: “AAPH group only added AAPH."			This is the list of corrections of our manuscript entitled ‘Study on the Antioxidant Properties Enhancement of Rice Bran by Mixed Bacteria Solid-state Fermentation (No.			fermentation8050212_perova	0
+Response 1: It is our great pleasure to get your professional comments.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_makarova	0
+The references cited are relevant and current, mostly published within the last three years.			The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+The conclusions are consistent with the presented results of the study.			The conclusions are consistent with the presented results of the study.			gels8040212_makarova	0
+Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			Response 18: Thanks for your carefully check.			gels8040212_makarova	0
+The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			The conclusions are consistent with the presented results of the study.			gels8040212_makarova	0
+We take the concerns seriously and comments have been carefully addressed.			This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			gels8040212_makarova	0
+Response 1: It is our great pleasure to get your positive comments.			This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			gels8040212_makarova	0
+Author Response Point 1: This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			gels8040212_makarova	0
+We take the concerns seriously and comments have been carefully addressed.			Thanks very much for your responsible and efficient work in the process of our manuscript.			gels8040212_makarova	0
+The article is very well written and only requires few minor changes.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_makarova	0
+The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_makarova	0
+Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			The conclusions are consistent with the presented results of the study.			gels8040212_makarova	0
+Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			What equipment was used for the texture profile analysis?			gels8040212_makarova	0
+This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+One main request would be to make it clearer what was the formulations and conditions tested.			Figure 2 has been modified accordingly to make the legend clearer.			gels8040212_makarova	0
+One main request would be to make it clearer what was the formulations and conditions tested.			One main request would be to make it clearer what was the formulations and conditions tested.			gels8040212_makarova	0
+The manuscript is presented in a well-structured manner.The references cited are relevant and current, mostly published within the last three years.The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_makarova	0
+B1, B2, and B3 looks biphasic, was this the case?			Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			gels8040212_makarova	0
+Line 512: Was 1% found to be within the viscoelastic region?			The article is very well written and only requires few minor changes.			gels8040212_makarova	0
+The article is very well written and only requires few minor changes.			Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			gels8040212_perova	0
+Author Response Point 1: This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			Author Response Point 1: The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			gels8040212_perova	0
+This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			Figure 7-B: I suggest choosing just one shear rate, instead of providing both graphs one can provide the numerical values at a fixed shear rate Line 474: What is meant by optimize condition?			gels8040212_perova	0
+Response 18: Thanks for your carefully check.			The article is very well written and only requires few minor changes.			gels8040212_perova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			Line 500: Define PDI at first use Line 507: Clearance height or gap`?			gels8040212_perova	0
+Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			The article is very well written and only requires few minor changes.			gels8040212_perova	0
+Author Response Point 1: The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			Line 500: Define PDI at first use Line 507: Clearance height or gap`?			gels8040212_perova	0
+Line 60: define WPI at first use Line 84-88: This is a summary of what was carried out and not a discussion, please remove Figure 1: Include in Fig.1c what was the the % of WPI used Figure 2: Increase the font of the legend Line 166: English check Figure 3: rename using latin alphabet Figure 4: Increase legend as it is not visible.			Author Response Point 1: The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			gels8040212_perova	0
+Author Response Point 1: The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_perova	0
+B1, B2, and B3 looks biphasic, was this the case?			This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			gels8040212_perova	0
+Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels.			Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			gels8040212_perova	0
+The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			The obtained emulsion gels are well characterized in terms of their structure and rheological and mechanical properties.			gels8040212_perova	0
+The manuscript is presented in a well-structured manner.			The conclusions are consistent with the presented results of the study.			gels8040212_perova	0
+The results of the manuscript are reproducible using the information in the methods section, but some minor explanations are missing.			This article compares the production of emulsion gels by two different methods, namely heat-and TGase- induced emulsion gels.			gels8040212_perova	0
+The manuscript is presented in a well-structured manner.			The references cited are relevant and current, mostly published within the last three years.			gels8040212_perova	0
+This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			This section is a bit confusing so I would suggest authors include a Table with all the sample formulations/conditions tested.			gels8040212_perova	0
+B1, B2, and B3 looks biphasic, was this the case?			Line 478 and 485: Are the gel continuous phase and the protein-emulsion mixture the same thing?			gels8040212_perova	0
+One main request would be to make it clearer what was the formulations and conditions tested.			The article is very well written and only requires few minor changes.			gels8040212_perova	0
+The article is very well written and only requires few minor changes.			The aim of this work is to prepare emulsion gels based on whey protein isolate and gelatin by two different methods: an enzymatic and a physical method.			gels8040212_perova	0
+The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_perova	0
+The conclusions are consistent with the presented results of the study.			Droplet size and gelatin concentration are the factors that most affect the structure and texture of emulsion gels..The results of this study are important for structuring emulsion gels and adjusting the texture of food products.			gels8040212_perova	0
+Here, we focused on the biological function and potential molec- 20 ular mechanism of the response of IbmiR319a to drought stress in sweet potato.			Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier.			genes13030404_makarova	0
+We also found that the lignin content was reduced, which 25 led to increased brittleness in MIM319.			This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.			genes13030404_makarova	0
+RNA-Seq results need to be further discussed.			34 35 1. Introduction 36 Drought is a major environmental factor causing abiotic stress (Pearce and RS 2001, 37 Xie, Wang et al. 2017, Meng, Li et al. 2018).			genes13030404_makarova	0
+16 Abstract: MicroRNA319 (miR319) plays a key role in plant growth, development, and multiple re- 17 sistance by repressing the expression of targeted TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) 18 genes.			Ren et al.'s study focus on the characterization of IbmiR319 based on its effect on plant architecture and drought response.			genes13030404_makarova	0
+34 35 1. Introduction 36 Drought is a major environmental factor causing abiotic stress (Pearce and RS 2001, 37 Xie, Wang et al. 2017, Meng, Li et al. 2018).			Author Response Please see the attachment Author Response File: Author Response.docx			genes13030404_makarova	0
+Taken together, our findings suggest that IbmiR319a affects plant architecture 32 by targeting IbTCP11/17 to control the response to drought stress in sweet potato.			There are some small edits that need addressing for clarity.			genes13030404_makarova	0
+It is 39 therefore essential that crops with drought tolerance traits are produced.			There are some small edits that need addressing for clarity.			genes13030404_makarova	0
+45 Citation: Lastname, F.; Lastname, F.; Lastname, F. Title.			Here, we focused on the biological function and potential molec- 20 ular mechanism of the response of IbmiR319a to drought stress in sweet potato.			genes13030404_makarova	0
+Here authors capture the early response to PEG compared to the drought response (water withdrawal) assessed earlier.			This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.			genes13030404_perova	0
+It is 39 therefore essential that crops with drought tolerance traits are produced.			16 Abstract: MicroRNA319 (miR319) plays a key role in plant growth, development, and multiple re- 17 sistance by repressing the expression of targeted TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) 18 genes.			genes13030404_perova	0
+As the authors briefly mentioned, PEG stress is more severe than water withdrawal, so it will not give a solid verification provided there are two variables, inducer, and timing.			It is 39 therefore essential that crops with drought tolerance traits are produced.			genes13030404_perova	0
+This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.			Taken together, our findings suggest that IbmiR319a affects plant architecture 32 by targeting IbTCP11/17 to control the response to drought stress in sweet potato.			genes13030404_perova	0
+Taken together, our findings suggest that IbmiR319a affects plant architecture 32 by targeting IbTCP11/17 to control the response to drought stress in sweet potato.			This is essential given that no temporal expression baseline is established for IbTCP11/17 under water withdrawal stress and the presence of other regulatory mechanisms of IbTCP11/17.			genes13030404_perova	0
+There are some small edits that need addressing for clarity.			Please rephrase and indicate what other processes are being referred to.			genes13030404_perova	0
+"""sequester the normal expression"" is nonsensical."			Response to Reviewer 1 Comments Point 1: The authors look at the expression of IbTCP11/17 (targets of miR319) from lines 418-424.			genes13030404_perova	0
+Genes 2022, 13, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Last- name Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations.			The authors addressed some of the suggestions made.			genes13030404_perova	0
+The manuscript is well written and the presented evidence supports the conclusions being made.			While congratulating the authors on this interesting study, I have the following comments.			genes13030404_perova	0
+Clean up the legend: 1. a) Remove underscores,done 2. b) Correct use of small caps, large caps in labels.done 3. c) Correct spelling (e.g. frameworkboundaries to framework boundaries; populated places to Populated places/(Settlements?); PossibleFluidEscapeFeatures to Possible fluid-escape features etc. ).			Reviewer 1: The authors present an interesting case study integrating geophysical, geological and hydrochemical data to map potential pathways for water and gases across complex sedimentary formations in the Northern Territories, in Australia.			geosciences12010037_makarova	0
+done Figure 1B -Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin.			This output was then overlayed upon satellite and magnetic images where lineaments are evident for checking and quality control.			geosciences12010037_makarova	0
+- Line352: “However, the elevated helium concentrations show no systematic pattern of increase with flow distance (as expected for in-situ production along the flow path) but a rather patchy regional pattern (Figure 8).” This should refer to Figure 9.done -Line 582.			Author Response The authors would like to thank you the reviewer 2 for this second round of reviews.			geosciences12010037_makarova	0
+Interpretation of shallow horizons and faults using all publicly available geological and geophysical data of the Beetaloo region.			Helium isotope data do not provide an unequivocal answer to whether the gas originates from the unconventional plays or a deeper source.			geosciences12010037_makarova	0
+the seismic interpretation of figure 7G (cross) that were misleading the reader and add the interpretation of the fluid leakage features on the images • Figure 9: Remove ‘s’ from <10 yearss.			We may mention that in the text - but for such a signature to be discernible the bulk of the water needs to be geothermal, and what we see is only a small admixture.			geosciences12010037_makarova	0
+Figures, however, need to be significantly improved for the final manuscript (see detailed comments below).			Detailed comments Aeromagnetic survey data acquired over the last 50+ years were employed for mapping structural and lithological features.			geosciences12010037_makarova	0
+: Fault related fluid flow implications for unconventional hydrocarbon development, Beetaloo Sub-basin (Northern Territory, Australia) Summary of content: The study investigates potential connections between unconventional petroleum plays and water assets in the Beetaloo Sub-basin in the Northern Territory, Australia.			Clean up the legend: 1. a) Remove underscores,done 2. b) Correct use of small caps, large caps in labels.done 3. c) Correct spelling (e.g. frameworkboundaries to framework boundaries; populated places to Populated places/(Settlements?); PossibleFluidEscapeFeatures to Possible fluid-escape features etc. ).			geosciences12010037_makarova	0
+Why are these not classified in terms of signal quality?			Where does the fourth category, “Fair”, which also is a qualitative category, fit in, and why not include this category in the legend?			geosciences12010037_makarova	0
+If these items are present, please use a more contrasting signatures to make it more visible.			Review summary Scientific aims clearly stated.			geosciences12010037_makarova	0
+The authors present an interesting case study integrating geophysical, geological and hydrochemical data to map potential pathways for water and gases across complex sedimentary formations in the Northern Territories, in Australia.			- Line 303: “We reviewed the seismic data in detail at the intersections with north-northwest trending lineaments observed on the magnetic dataset (Figure 4).” No such lineaments are shown in Figure 4.			geosciences12010037_makarova	0
+The depth maps provided in Figure 5 should be supplemented by isochore maps for the rock volumes between pairs of mapped reflectors.			To my understanding this should correspond to Figure 2 a, which shows substantially more than 130 lineaments.			geosciences12010037_makarova	0
+These often highlight tectonic accommodation space creation better than maps.			Recent fault activity was identified using a combination of sub-surface imaging and comparison of fault orientations and surface drainage networks.			geosciences12010037_makarova	0
+One of the key issues in the Beetaloo Sub-basin, the risk of groundwater contamination from fracking-based production of the unconventional plays.			All comment avec been considerate: figures 1,4,7,8, 9 have been			geosciences12010037_makarova	0
+); PossibleFluidEscapeFeatures to Possible fluid-escape features etc.			We added: “We define as recent tectonic activity the events that are recorded at the surface and can alter the surface with active tectonic features, such as modifications of the drainage system” • Discussion: One general comment, could the use of other tracers (radon, of stable isotopes of water) yield also insight on the origin of the fluids sampled in this study?			geosciences12010037_makarova	0
+According to the new caption shows the unfiltered surface drainage network.			Consider removing labels not referred to in the text.			geosciences12010037_makarova	0
+Overall, the implications of this study are important not only for industrial purposes (reservoir exploration and operation) but also for our basic understanding of subsurface flow processes over geological timescales.			Consider using higher contrast colours or thicker lines to highlight different faults on the map.			geosciences12010037_makarova	0
+Development of the petroleum resources in this region is controversial and faces opposition from Traditional Owners as well as climate scientists, and environmentalists.			Conclusions appear supported by the observations made in this study, but presentation of results needs to be improved.			geosciences12010037_makarova	0
+However, there are still a couple of minor issues.			Helium content of the aquifer systems was sampled and measured.			geosciences12010037_makarova	0
+Methods are sound and clearly described, There are some shortcomings in documentation and presentation of results, which weaken the link between observations and conclusion.			Magnetic data and 2D seismic reflection profiles were used to image structural features, and fluid/gas leakage pathways.			geosciences12010037_makarova	0
+These often highlight tectonic accommodation space creation better than maps.			We separated the legends from figures 1A and 1B as to add clarity.			geosciences12010037_makarova	0
+Not possible to differentiate “ntspr_2M_gw” (whatever that is) and “PossibleFluidEscapeStructures” (same colour).			Helium isotope data do not provide an unequivocal answer to whether the gas originates from the unconventional plays or a deeper source.			geosciences12010037_makarova	0
+This figure could be improved by adding shaded satellite image to help the reader compare the mapped drainage systems (which were automatically mapped) and fault traces/lineaments with the real features observed.			If these items are present, please use a more contrasting signatures to make it more visible.			geosciences12010037_makarova	0
+We tried to improve the figures at best- attached is a detailed reply to reviews.			It doesn’t read well otherwise.done • Figure 7: What is special about the locations highlighted by the markers (crosses on Figure 7B and square markers on Figure 7C).			geosciences12010037_makarova	0
+Methods are sound and clearly described, There are some shortcomings in documentation and presentation of results, which weaken the link between observations and conclusion.			The approach used by the authors is innovative and allows to elaborate several hypotheses regarding the occurrence and movement of both paleo and more recent fluids.			geosciences12010037_makarova	0
+Alternatively the position of Figure 6 A, B and C should be indicated in Figure 4 as well as Figure 3. done Figure 5.			Review summary Scientific aims clearly stated.			geosciences12010037_makarova	0
+We followed those advises and updated the document consequently.			One of the key issues in the Beetaloo Sub-basin, the risk of groundwater contamination from fracking-based production of the unconventional plays.			geosciences12010037_makarova	0
+All comment avec been considerate: figures 1,4,7,8, 9 have been			Figure 2 - Poor quality figure (low resolution) in the version available to the reviewer.			geosciences12010037_makarova	0
+The present paper is thus part of an ongoing debate and the findings presented here are likely to influence discussions and decisions by stakeholders.			• Lines 271-272: Why is the signal quality so variable, and how were the categories (excellent, average and poor) defined?			geosciences12010037_makarova	0
+The replies to detailed comments are in purples, directly below each comment Frery et al.			Fault and surface drainage network mapping (tectonic geomorphology).			geosciences12010037_makarova	0
+Outline size modified and homogenised with other figures 4.			With respect to the helium measurements, no detailed documentation of where these samples were collected is provided beyond referring to Geological and Bioregional Assessment Program (2021a) Fact sheet 12, and Geological and Bioregional Assessment Program (2021b) Regional tracer results from the Cambrian Limestone Aquifer.			geosciences12010037_makarova	0
+what differentiates “FAULT” from a “BaseCambrianSeismic fault”?).			Why are these not classified in terms of signal quality?			geosciences12010037_makarova	0
+It is not clear if the surface drainage network line dataset is based on the “unfiltered” or “simplified” data shown in Figure 2.			Thank you, the details comments on those figures have been followed and new figures are provided Helium measurements from groundwater samples.			geosciences12010037_makarova	0
+The study concludes that there the shallow aquifers are connected to deep-seated gas source along faults, some of which show evidence of recent movement.			• Figure 6: Put the title of each subfigure above the plot.			geosciences12010037_makarova	0
+Reference is made to “…prominent […] magnetic lineaments which can be caused by faulting”, and “…several circular low magnetization anomalies, approximately two kilometers in diameter, aligned with northwest trending lineaments”.			done -Many well-name labels are hard difficult to read/illegible.			geosciences12010037_makarova	0
+done -Many well-name labels are hard difficult to read/illegible.			If the latter applies, make sure to provide a scale.			geosciences12010037_makarova	0
+It would be nice to include a map with these features and lineaments interpreted from the aeromagnetic datasets.			Reviewer 2: Summary of content: The study investigates potential connections between unconventional petroleum plays and water assets in the Beetaloo Sub-basin in the Northern Territory, Australia.			geosciences12010037_makarova	0
+done Figure 4 -Poor quality figure (low resolution) in the version available to the reviewer.			The depth maps provided in Figure 5 should be supplemented by isochore maps for the rock volumes between pairs of mapped reflectors.			geosciences12010037_makarova	0
+done Figure 1B -Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin.			the seismic interpretation of figure 7G (cross) that were misleading the reader and add the interpretation of the fluid leakage features on the images • Figure 9: Remove ‘s’ from <10 yearss.			geosciences12010037_makarova	0
+Depth conversion using check-shot velocities from 26 wells.			done Figure 1B -Consider adding a small inset table or schematic to showing the Cambrian Limestone Aquifer stratigraphy in the Georgina, Daly and Wiso basin.			geosciences12010037_makarova	0
+Helium content of the aquifer systems was sampled and measured.			what differentiates “FAULT” from a “BaseCambrianSeismic fault”?).			geosciences12010037_makarova	0
+The results are of interest to both surgeons and haematologists/haemostaseologists.			For example, the section begins with introducing inherited thrombophilias and VTE risk in these conditions, then proceeds to surgery, then back again to thrombophilia where it repeats itself and even states other numbers for prevalences and VTE risks than previously stated.			healthcare10030476_makarova	0
+Please explain, or consider omitting this statement altogether.			Answer to the Reviewer Point 1: The observation of the reviewer has been accepted and the table 1 was modified accordingly.			healthcare10030476_makarova	0
+The results are of interest to both surgeons and haematologists/haemostaseologists.			2-3% of what for the 20210 polymorphism and 3-8% of what for FV Leiden?			healthcare10030476_makarova	0
+Please explain, or consider omitting this statement altogether.			Point 4: In the Materials and methods section, several questions arise: A)			healthcare10030476_makarova	0
+Answer to the Reviewer point 7: The observation of the reviewer has been accepted and the manuscript was modified accordingly.			However, I have several concerns which must be adequately addressed before the manuscript is suitable for publication.			healthcare10030476_makarova	0
+Answer to the Reviewer Point 1: The observation of the reviewer has been accepted and the table 1 was modified accordingly.			We also declare that we have no conflict of interest in connection with this paper.			healthcare10030476_perova	0
+Author Response Reviewer(s)' Comments to Author: Reviewer 2: (x) Extensive editing of English language and style required Answer to the Reviewer: The new manuscript has been evaluated by an expert of English language.			D) Page 3, lines 140-141: “We collected data for all groups of patients in terms of drain permanence, thromboembolic events, and hematoma formation.” How did you define and verify a thromboembolic event?			healthcare10030476_perova	0
+Answer to the Reviewer Point 2: The new manuscript has been evaluated by an expert of English language.			Point 5: The Discussion section could benefit from a language revision, especially in the latter part, to Answer to the Reviewer point 5: The observation of the reviewer has been accepted and the manuscript was modified accordingly.			healthcare10030476_perova	0
+2-3% of what for the 20210 polymorphism and 3-8% of what for FV Leiden?			"The authors report experience from their surgical center with performing abdominoplasty in women with ""thrombophilia"" which is not clearly defined."			healthcare10030476_perova	0
+We thank the Editor and the Reviewers for helping us to improve our paper.			Point 2: Also, another round of language editing may be suitable.			healthcare10030476_perova	0
+I think, the manuscript benefits from a minor language editing.			Also, the research questions lack describing the study setting.			healthcare10040723_perova	0
+I suggest to remove the phrase ‘so called,’ that has been used in the conclusion section or any other section.			For example, the first sentence should describe the general aim or the overall objective of the study in a more general term.			healthcare10040723_perova	0
+However, the first sentence did not appear to serve this purpose.			Page 3, lines 117-118, the statement has two issues.			healthcare10040723_perova	0
+I suggest to re-write the aims and study questions more clearly.			I would ask, why did you use convenience sampling?			healthcare10040723_perova	0
+However, the first sentence did not appear to serve this purpose.			It needs the careful wording of the study participants.			healthcare10040723_perova	0
+Page 4, table 1, the caption and the table content do not match.			‘The items were then tested through a pilot study consisting of a 117 group of 4 pre- and clinical students, mediated by AG and JE’.			healthcare10040723_perova	0
+I suggest to re-write the aims and study questions more clearly.			I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.			healthcare10040723_perova	0
+It needs the careful wording of the study participants.			Also, the research questions lack describing the study setting.			healthcare10040723_perova	0
+However, the first sentence did not appear to serve this purpose.			Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement.			healthcare10040723_perova	0
+I am suggesting this because the logic of quantitative sampling and qualitative sampling are different.			I suggest the authors to replace this with appropriate phrase.			healthcare10040723_perova	0
+I think, the manuscript benefits from a minor language editing.			Also, the research questions are not clearly stated.			healthcare10040723_perova	0
+Around 38% of the students felt ill-prepared and could not answer AI-related questions because digitization in medicine and AI are not a formal part of the medical curriculum.			1 Review comments Title: Medical students' perceptions towards digitization and artificial intelligence: A mixed methods study Thank you for the opportunity to review this manuscript.			healthcare10040723_perova	0
+Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement.			Page 3, lines 117-118, the statement has two issues.			healthcare10040723_perova	0
+It needs the careful wording of the study participants.			Also, the research questions lack describing the study setting.			healthcare10040723_perova	0
+In the abstract, lines 23-27, I suggest to use the following expression or something like that.			Page 12, line 389, the statement which says, ‘The non-probabilistic sampling makes a generalization of the answers difficult,’ is confusing.			healthcare10040723_perova	0
+Page 4, table 1, the caption and the table content do not match.			I hope these suggestions are of use to the authors.			healthcare10040723_perova	0
+Dear Authors, Here attached, please find my comments for the improvement of your paper.			The paper is well written in terms of clarity of idea and the authors use of mixed-methods design was interesting.			healthcare10040723_perova	0
+Page 13, lines 409-410, in the conclusion section, the statement which says, ‘This study also found significant differences between those groups indicating differences in subgroups of students from the quantitative survey.’ This is a confusing statement.			The aim of the study as reported in the manuscript was to explore attitudes of 2020 medical students’ generation towards various aspects of eHealth technologies with the focus on AI using an exploratory sequential mixed method analysis.			healthcare10040723_perova	0
+In the abstract, lines 23-27, I suggest to use the following expression or something like that.			I suggest to remove the phrase ‘so called,’ that has been used in the conclusion section or any other section.			healthcare10040723_perova	0
+Response 10: Special thanks to you for your good suggestion.			"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_makarova	0
+In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China).			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_makarova	0
+This sentence has been changed to “To better understand why OfSPL11 transgenic lines exhibit enhanced abiotic stress tolerance, we analyzed the expression of known abiotic stress response genes in 3-week-old wild-type and transgenic Arabidopsis.” Point 4: In lines 271-272 and 395-396, how do you explain that salt induced promoter trans-activation?			Response 4: We feel sorry for the unclear statement.			horticulturae8050412_makarova	0
+In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions.			"Point 11: In line 100 replace ""ligated"" with was ligated and quote a reference for pORER4-GFP vector or mention brand if it is commercial."			horticulturae8050412_makarova	0
+In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions.			The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves.			horticulturae8050412_makarova	0
+Point 4: Line 267-268: Please, check this sentence.			Therefore, we suggest that salt stress can induce the activity of the OfSPL11 promoter.			horticulturae8050412_makarova	0
+"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			"Point 9: Line 360: remove the space after ""assay""."			horticulturae8050412_makarova	0
+"Point 14: In line 118 define ""symbiotic medium""."			"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			horticulturae8050412_makarova	0
+Results clearly describe the protein localization as well as its role in stress response.			In a previous study (Ding, 2021), the LlWRKY39 promoter was fused to a pGreenII-0800-LUC vector using a recombinant ligase (Vazyme, Nanjing, China).			horticulturae8050412_makarova	0
+"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			The LUC signal increased profoundly after subjection to HS (37℃), although under normal conditions (room temperature), the LUC signal was very low, which implied that HS activated the promoter activity of LlWRKY39.			horticulturae8050412_makarova	0
+The constructed vector and an empty vector (control) were transformed into A. tumefaciens strain GV3101 (pSoup), which was used to transform N. benthamiana leaves.			Results clearly describe the protein localization as well as its role in stress response.			horticulturae8050412_makarova	0
+"Point 23: In line 262 replace ""were treatment by"" with were treated with."			; Zhang, D.H.; Teng, N.J. LlWRKY39 is involved in thermotolerance by activating LlMBF1c and interacting with LlCaM3 in lily (Lilium longiflflorum).			horticulturae8050412_makarova	0
+Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			horticulturae8050412_makarova	0
+In this study, the experimental operations, reagents and plant materials were were the same as the previous study, except for the different treatment conditions.			"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_makarova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_makarova	0
+In this view, I've only some few questions for the authors.			tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 5:			horticulturae8050412_makarova	0
+Response 9: Thank for your kindly suggestion.			Results clearly describe the protein localization as well as its role in stress response.			horticulturae8050412_makarova	0
+"Point 7: In line 65 replace ""And"" with In addition."			"Point 25: In line 292 ""The"" must be lowercase."			horticulturae8050412_makarova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			horticulturae8050412_makarova	0
+Response 5: Thank for your kindly suggestion.			The paper by Zhu and coworkers deal with the functional deeeping on te role of gene OfSPL11 from Osmanthus fragrant.			horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_makarova	0
+"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			Results clearly describe the protein localization as well as its role in stress response.			horticulturae8050412_makarova	0
+"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			In this view, I've only some few questions for the authors.			horticulturae8050412_makarova	0
+Do the authors have performed an efficiency test before using it?			"Point 7: In line 65 replace ""And"" with In addition."			horticulturae8050412_makarova	0
+The LUC signal increased profoundly after subjection to HS (37℃), although under normal conditions (room temperature), the LUC signal was very low, which implied that HS activated the promoter activity of LlWRKY39.			We have revised the manuscript according to your comments (Line 274-277).			horticulturae8050412_makarova	0
+Results clearly describe the protein localization as well as its role in stress response.			We have revised the manuscript according to your comments (Line 274-277).			horticulturae8050412_makarova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			Results clearly describe the protein localization as well as its role in stress response.			horticulturae8050412_makarova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			"Point 25: In line 292 ""The"" must be lowercase."			horticulturae8050412_makarova	0
+Point 17: In line 131 delete the period after experiments.			"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+Therefore, we suggest that salt stress can induce the activity of the OfSPL11 promoter.			"It ends with ""then"" Response 4: Thank you for your comment."			horticulturae8050412_makarova	0
+Point 7: Line 311: plant leaves instead of plants leaves.			Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_makarova	0
+Point 4: Line 267-268: Please, check this sentence.			"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			horticulturae8050412_makarova	0
+"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			A deeeping on protein an nucleotide structures was also reported.			horticulturae8050412_makarova	0
+We have revised the manuscript according to your comments (Line 274-277).			"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			horticulturae8050412_makarova	0
+"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			"Line 315: maybe a semicolon after ""salt stress"" is appropriate"			horticulturae8050412_makarova	0
+Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_makarova	0
+; Bustin, S.A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR.			"Point 6: In line 63 replace ""trancroptoma"" with transcriptome."			horticulturae8050412_perova	0
+"Point 9: Line 360: remove the space after ""assay""."			We have revised the manuscript according to your comments (Line 274-277).			horticulturae8050412_perova	0
+"Point 27: In line 395 replace ""it revealed"" with revealed."			"Point 26: In line 318 ""We"" must be lowercase."			horticulturae8050412_perova	0
+; Zhang, D.H.; Teng, N.J. LlWRKY39 is involved in thermotolerance by activating LlMBF1c and interacting with LlCaM3 in lily (Lilium longiflflorum).			"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			horticulturae8050412_perova	0
+Furthermore, I reccomend a text revision since some errors are present in the main text.			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_perova	0
+"Point 27: In line 395 replace ""it revealed"" with revealed."			Response 9: Thank for your kindly suggestion.			horticulturae8050412_perova	0
+"Point 16: In line 129 replace ""strains and ""strain"" with lines and line."			Response 4: We feel sorry for the unclear statement.			horticulturae8050412_perova	0
+"Point 24: In line 264 replace ""indicate"" with indicated."			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_perova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			horticulturae8050412_perova	0
+Point 28: In line 406 delete period after stress.			Identification of Suitable Reference Genes for Gene Expression Normalization in the Quantitative Real-Time PCR Analysis of Sweet Osmanthus (Osmanthus fragrans Lour.).			horticulturae8050412_perova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			"Point 23: In line 262 replace ""were treatment by"" with were treated with."			horticulturae8050412_perova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			horticulturae8050412_perova	0
+Point 10: Figure 9 caption: measure instead of measuring.			Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_perova	0
+"Point 26: In line 318 ""We"" must be lowercase."			Point 10: Figure 9 caption: measure instead of measuring.			horticulturae8050412_perova	0
+"Point 6: In line 63 replace ""trancroptoma"" with transcriptome."			Response 3: Special thanks to you for your good suggestion.			horticulturae8050412_perova	0
+"Point 7: In line 65 replace ""And"" with In addition."			Response 7: Special thanks to you for your good suggestion.			horticulturae8050412_perova	0
+"Point 12: Line 446: ""..participate in salt stress"" Response 12: Thank for your kindly suggestion."			"Point 24: In line 264 replace ""indicate"" with indicated."			horticulturae8050412_perova	0
+"Point 23: In line 262 replace ""were treatment by"" with were treated with."			; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S.			horticulturae8050412_perova	0
+"Point 9: Line 360: remove the space after ""assay""."			"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_perova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			horticulturae8050412_perova	0
+Major corrections The manuscript by Zhu et al., describes the cloning of OfSPL1 transcriptional activator from Osmanthus fragans, its molecular characterization, subcellular localization, gene-expression, trans-activation in tobacco cells and two-hybrid system, and overexpression in Arabidopsis, displaying and enhanced salt-tolerance phenotype and protection from oxidative damage.			"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			horticulturae8050412_perova	0
+Response to Reviewer 2 Comments Point 1: Why the authors do not consider to use aminoacidic sequence to compute a protein model (e.g., by using gromacs)?			"Point 12: Line 446: ""..participate in salt stress"" Response 12: Thank for your kindly suggestion."			horticulturae8050412_perova	0
+; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S.			"Point 12: Line 446: ""..participate in salt stress"" Response 12: Thank for your kindly suggestion."			horticulturae8050412_perova	0
+; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S.			"Point 8: Line 315: maybe a semicolon after ""salt stress"" is appropriate."			horticulturae8050412_perova	0
+In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			In addition, heterologous overexpression promotes induction of several genes involved in tolerance to abiotic stress.			horticulturae8050412_perova	0
+"Point 22: In line 261 replace ""tobaccos"" with tobacco plants."			Point 7: Line 311: plant leaves instead of plants leaves.			horticulturae8050412_perova	0
+"Point 12: In line 113 replace ""psoup"" with pSOUP and ""tobaccos"" with tobacco plants."			"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_perova	0
+This goal was achieved using transgenic lines to investigate the role of this gene in salt stress response.			"Point 14: In line 118 define ""symbiotic medium""."			horticulturae8050412_perova	0
+"Response to Reviewer 1 Comments Point 1: ""back of tobacco"" is not clear be more specific."			"Point 11: In line 100 replace ""ligated"" with was ligated and quote a reference for pORER4-GFP vector or mention brand if it is commercial."			horticulturae8050412_perova	0
+"Point 7: In line 65 replace ""And"" with In addition."			tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 5: Figure 5 caption: activates instead of activate.			horticulturae8050412_perova	0
+2015, 10,1-17. https://doi.org/10.1371/journal.pone.0136355 Point 3: Line 124: generat instead of generate.			"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			horticulturae8050412_perova	0
+In Silico and Transcription Analysis of Trehalose-6-phosphate Phosphatase Gene Family of Wheat: Trehalose Synthesis Genes Contribute to Salinity, Drought Stress and Leaf Senescence.			; Zhao, K.; Wang, S.; Sikdar, A.; Noor, H.; Jeon, J.S.			horticulturae8050412_perova	0
+"Point 11: Line 392: I suggest to change the sentence in the following way"".. to drive the expression of target genes.."" Response 11: Thank you for your comment."			In this view, I've only some few questions for the authors.			horticulturae8050412_perova	0
+"2021, 8,36. https://doi.org/10.1038/s41438-021-00473-7 Point 5: In line 38 replace ""Fortunately"" with However."			; Zhang, D.H.; Teng, N.J. LlWRKY39 is involved in thermotolerance by activating LlMBF1c and interacting with LlCaM3 in lily (Lilium longiflflorum).			horticulturae8050412_perova	0
+"Point 27: In line 395 replace ""it revealed"" with revealed."			The paper by Zhu and coworkers deal with the functional deeeping on te role of gene OfSPL11 from Osmanthus fragrant.			horticulturae8050412_perova	0
+Point 28: In line 406 delete period after stress.			tumefaciens suspensions containing the fusion vector OfSPL11pro: LUC was transiently transformed into O. fragrans callus using vacuum infiltration, and then the O. fragrans callus were cultured in a symbiotic medium containing 0 mM and 200 mM NaCl.” Point 5: Figure 5 caption: activates instead of activate.			horticulturae8050412_perova	0
+However, we used the data without outlier to construct the figure.			How much soil (kg) did you collect for each sample?			ijerph19127332_makarova	0
+Table 1 – is it possible to list/ summarise the data for each day?			The results provided insights into the varying health risks from different have metals.			ijerph19127332_makarova	0
+In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn.			Please, use element symbols not the entire name in all the manuscript.			ijerph19127332_makarova	0
+Page 3, lines 121-124:Please, add some information: How did you collect dust samples? How much soil (kg) did you collect for each sample?			Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue.			ijerph19127332_makarova	0
+Explain for better understanding Answer: For this question, our answer is following: this data is from the Technical Guidelines for Soil Pollution Risk Assessment of Construction Land in China.			Answer: Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C).			ijerph19127332_makarova	0
+At last, we also added the heavy metals information in resident or park green areas: “In resident and park green areas, the highest concentration of heavy metals is Zn and the lowest is Cd.			Title- ‘in’ should be used for larger areas, can you add the study locations?			ijerph19127332_makarova	0
+Answer: Thanks for your advice, we have added the specific parameters and modified them as follows:” When the collection was completed, the samples were taken back to the laboratory and stored first in the refrigerator (‒20°C).			Explain for better understandingWill like to see ethical approval for the project/research?			ijerph19127332_makarova	0
+Title- ‘in’ should be used for larger areas, can you add the study locations?			Explain for better understandingWill like to see ethical approval for the project/research?			ijerph19127332_makarova	0
+Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas.			Line 368 – what is the meaning of ‘Whatever’?			ijerph19127332_makarova	0
+I’d suggest some modifications to improve the paper.			Page 3, lines 131-151: Please, add some information:How many millilitres of HNO3, H2O2, HF, HClO4 did you use for the digestion procedures (food samples, faces samples, soil and dust samples)?			ijerph19127332_makarova	0
+But we failed to get all children’s feces and urine sample every day.			Thus, we cannot provide the data for each day.			ijerph19127332_makarova	0
+S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”.			It has been changed to “The LRL is ‒112.4‒100.9 mg/d which is considered as the lower limit of the 90% confidence interval (CI) of the 2.5th percentile (P2.5), whereas the URL is 516.9‒730.2 mg/d, which is considered as the upper limit of the 90% CI of the 97.5th percentile (P97.5).			ijerph19127332_makarova	0
+Lines 253 and 377: different values between SIR and figure 3.			Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_makarova	0
+26 Keywords: E‒waste; Heavy metal pollution; Children; Soil and dust ingestion rates; Health risk 27 assessment 28 29 1.			The median (minimum and maximum) of SIR values were ‒124.3 (‒278.0 to 228.2), ‒210.2 (‒490.1 to 273.8), 27.1 (0.4 to 106.0), ‒22532.8 (‒29443.8 to ‒6215.9), 23.9 (‒45.3 to 268.0), 175.3 (‒56.4 to 1040.7), 39.2 (‒36.4 to 284.0), and ‒263.2 (‒491.4 to 132.3) mg/d for Al, Ba, Ce, Mn, Sc, Ti, Y, and V.” Point 6.			ijerph19127332_makarova	0
+The concentrations of heavy metals in park green areas were slightly higher than that in residential area, which may attribute to the difference of soil environmental quality management and control policy between these areas.			I would like to see a better description of the study location.			ijerph19127332_makarova	0
+Integrated data from the resident and park green areas, the median concentrations (mg/kg) of Cr, Ni, Cu, Zn, As, Cd, and Pb were 48.8, 63.9, 128, 413, 6.30, 0.513, and 115, respectively.” Author Response File: Author Response.docx			- why different processing for different samples?			ijerph19127332_makarova	0
+These findings provide important insights into the exposure risks of heavy metals in e‒ 25 waste dismantling sites and emphasize the health risk of Cr and As.			We have added the description of sampling site in the materials and methods.			ijerph19127332_makarova	0
+Besides, our study focuses on estimating the children’s SIRs of from e‒waste disassembly areas by collecting and analyzing selected tracer elements in matched samples of their consumed food, feces, and urine, as well as soil samples from their play areas.			Please, check captions of figures S2 and 3 that are identical.			ijerph19127332_makarova	0
+Experimental factors such as measurement error, source error, and transit time misalignment, may lead to some of these outliers.			And then samples were microwaved with HNO3‒H2O2.			ijerph19127332_makarova	0
+Lines 116-118: The amounts of samples were confused.			Dried samples (1 g) were digested to evaporate at low temperatures of 55°C on a heating plate with 3 mL concentrated nitric acid, 3 mL hydrogen fluoride, and 1 mL perchloric acid (HNO3‒HF‒HClO4).			ijerph19127332_makarova	0
+Application of more biotic samples that you suggested is useful for our study, however, it is hard to get other samples from children, such as blood and tissue.			The medians of the SIR values after removing the high values (see Figure S3(b) and Figure 1) were taken as the final SIR values for children living in the studied e‒waste dismantling site in South China.” Point 7.			ijerph19127332_makarova	0
+I suggested the major revision before publication.			The title of Figure 1 has been changed to “Figure 1.			ijerph19127332_makarova	0
+As an example, they may analyze the influences of food intake and food intake on heavy metal exposure.			Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_makarova	0
+In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR.			Also describe in the methods sectionLine 368 – what is the meaning of ‘Whatever’?			ijerph19127332_makarova	0
+Answer: Thanks for your advice, and we have added references to support this view, it is added “[30] Huang, W.L.			Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			ijerph19127332_makarova	0
+At first, the soil heavy metal data has been added in the supporting information (Table S4 and S5).			Lines 176-178: Different units between manuscript and SM (figure S1).			ijerph19127332_makarova	0
+The results provided insights into the varying health risks of different have metals.			This study estimated the soil ingestion of 66 17 children from e‒waste disassembly areas by collecting and analyzing selected tracer elements in 18 matched samples of their consumed food, feces, and urine, as well as soil samples from their play 19 areas.			ijerph19127332_perova	0
+It has been re- 38 ported that the concentration of Cu was about three times higher than the Grade II guide- 39 line level (Soil Environmental Quality Standard, GB 15618‒1995) in an e‒waste disassem- 40 bly area in China, and the Cu concentrations of soils from dumping, burning, and acid 41 leaching sites were found to be 10, 40, and 60 higher, respectively, than the Grade II level.			It has been re- 38 ported that the concentration of Cu was about three times higher than the Grade II guide- 39 line level (Soil Environmental Quality Standard, GB 15618‒1995) in an e‒waste disassem- 40 bly area in China, and the Cu concentrations of soils from dumping, burning, and acid 41 leaching sites were found to be 10, 40, and 60 higher, respectively, than the Grade II level.			ijerph19127332_perova	0
+I suggested the major revision before publication.			We have examined the tracer elements in the food.			ijerph19127332_perova	0
+The conditions of microwave digestion are the same as mentioned above.			Title: Estimation of Children’s Soil and Dust Ingestion Rates and Health Risk in E Waste Dismantling Area Authors: Yan Yang * , Mengdi Zhang , Haojia Chen , Zenghua Qi , Chengcheng Liu , Qiang Chen , Tao Long Revisions Need:Title- ‘in’ should be used for larger areas, can you add the study locations?			ijerph19127332_perova	0
+Better explanation needed - why were the feces samples not microwaved?			Temperature of the heating plate for faces sample digestion.			ijerph19127332_perova	0
+In addition, children’s feces and urine belong to biological samples, which is sufficient to calculate the SIR.			Please, check captions of figures S2 and 3 that are identical.			ijerph19127332_perova	0
+In addition, dried soil samples from living spaces (0.5 g) were digested with the same pre-treated and analyzed by ICP‒MS for Pb, As, Cr, Cu, Ni, Cd, and Zn.			The digestion process is repeated until the sample becomes sticky.			ijerph19127332_perova	0
+Although there were 66 children involved in this study, 66 food samples, 62 feces and 64 urine samples were collected.			The conditions of microwave digestion are the same as mentioned above.			ijerph19127332_perova	0
+Lines 209 and 214: check the values of Al and Sc, because they were different among discussion, Table 1 and figure 1.			Finally, the digest was analyzed by HR‒ICP‒MS.			ijerph19127332_perova	0
+The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C.			And then samples were microwaved with HNO3‒H2O2.			ijerph19127332_perova	0
+Point 1、This study estimated the heavy metal exposure to children through soil and dust Ingestion in electronic waste (e-waste) dismantling areas.			Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			ijerph19127332_perova	0
+At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.			Dried soil and dust samples (0.5 g) were digested as same as feces and food samples on a heating plate by HNO3‒HF‒HClO4.			ijerph19127332_perova	0
+And then samples were microwaved with HNO3‒H2O2.			Sample Preparation and Instrumental Analysis -			ijerph19127332_perova	0
+The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y.			Human Body Burden of Heavy Metals and Health Consequences of Pb Exposure in Guiyu, an E-Waste Recycling Town in China.			ijerph19127332_perova	0
+At present, the related content in manuscript has been replaced to: “The SIR values based on the measured tracer element concentrations of food, feces, urine, soil and dust are presented in Table 1 and Figure 1.			The authors should provide detailed data about the concentration of heavy metals in soil and biotic samples.			ijerph19127332_perova	0
+Line 342- Describe with references the values for carcinogenic risk and non-carcinogenic risks in the methos sections before mentioning it in the discussion section.			Lines 176-178: Different units between manuscript and SM (figure S1).			ijerph19127332_perova	0
+The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS, Nu Attom, England) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y. Urine samples (15 mL) were placed in a digestion tube and digested by microwaving with 2 mL of H2O2 and 3 mL of concentrated HNO3.			I am satisfied with the revision made on the manuscript and would like to recommend the manuscript for publication.			ijerph19127332_perova	0
+Also describe in the methods section Answer: Thank you for your proposal.			Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			ijerph19127332_perova	0
+The conditions of microwave digestion are the same as mentioned above.			Page 3, lines 99-100: please, add references for the following information South China's economic conditions are better than in North China, but the environmental pollution is worse.			ijerph19127332_perova	0
+Finally, the digest was analyzed by HR‒ICP‒MS.			For this problem, we conducted a questionnaire survey when collecting samples before the experiment, and all the guardians of the sample providers knew and agreed to the experiment.			ijerph19127332_perova	0
+please rephrase Answer: Thanks for your advice, for the first question, this part of the ambiguous expression has been modified to “schools should keep desks and teaching AIDs clean and tidy, and urge children to clean up after outdoor activities” Point 11.			S3 Frequency distribution histogram and outlier box (a) and Frequency distribution histogram and outlier box (b) of soil ingestion rate (SIR) based on Al”.			ijerph19127332_perova	0
+Crushed Evaporated samples were microwaved with 2 mL concentrated nitric acid and 3 mL hydrogen peroxide (HNO3‒H2O2).			Better explanation needed Answer: Thanks for your suggestion, we have sorted out the method part again, and now the modification is as follows: “Feces samples and food samples were pretreated in the same way.			ijerph19127332_perova	0
+The supernatant was analyzed for Al, Ba, Mn, Ti, and V by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP‒OES, Spectra Arcos SOP, German) which is a method for atomic emission spectroscopy analysis using a light source that generates plasma discharge through high-frequency inductive coupling and by ICP‒MS for Ce, Sc, and Y.			The supernatant was extracted and analyzed by High Resolution Inductively Coupled Plasma Mass Spectrometry (HR‒ICP‒MS) to determine Mn, Al, Ba, Ti, Ce, V, Sc, and Y.			ijerph19127332_perova	0
+And then samples were microwaved with HNO3‒H2O2.			Page 3, lines 116 and 120: Please, add technical details about: lyophilization of food samples.			ijerph19127332_perova	0
+S4 Frequency distribution histogram and outlier box of soil ingestion rate (SIR) based on tracer Al(a), Ba(b), Ce(c), Mn(d), Sc(e), Ti(f), V(g), and Y(h) separately”.			The digested product was diluted to 30 ml with ultrapure water and then stored at 4°C.			ijerph19127332_perova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			1 Response to Reviewer 2 Comments Point 1: 2.			jcm11051247_makarova	0
+1 Response to Reviewer 2 Comments Point 1: 2.			Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.			jcm11051247_makarova	0
+The impact of Tocilizumab on radiological changes can not be adequately analyzed without a control group not receiving the drug.			1 Response to Reviewer 2 Comments Point 1: 2.			jcm11051247_makarova	0
+Results 86 cases are excluded from 187 cases, which is too many.			The median oxygen flow rate is shown to be 14 L/min, but FiO2 may vary depending on the method of oxygen delivery (nasal canulae, HNF, MV).			jcm11051247_makarova	0
+Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.			When comparing different oxygen delivery methods, isn't it more appropriate to compare FiO2 rather than oxygen flow rate?			jcm11051247_makarova	0
+1 Response to Reviewer 1 Comments Point 1: This study has a design flaw.			Point 2: The proposed timing of Tocilizumab administration based on oxygen flow rate is interesting but undoubtedly requires a more detailed statistical analysis to convince the readers.			jcm11051247_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			Study design and population If there are mutations or differences in viral strains during the course of the study, should differences in response to TOC and timing of administration depending on the strain be considered?			jcm11051247_makarova	0
+Thank you for improving this paper, especially regarding the timing of TOC administration. Still,			Timing of TOC administration according to the oxygen flow rates As mentioned above, the cutoff value is O2 13L/min, but the oxygen volume is difficult to interpret because FiO2 varies depending on the method of oxygen delivery.			jcm11051247_makarova	0
+Results 2 The oxygen flow rate of 13 L/min measured by ROC as a cutoff value for TOC administration should be presented by FiO2.			Comparative radiologic changes before and after TOC administration In Table 2, about the change in CT findings in fatal and non-fatal patients before and after TOC administration, isn't it the same as the difference in imaging findings between patients who survive after severe illness and those who do not, regardless of TOC?			jcm11051247_makarova	0
+The introduction part can highlight the key points and innovations of this paper.			It is recommended that the number of digits after the decimal point for the value of the correlation coefficient throughout the text be consistent.			land11030363_makarova	0
+Table 1: The pH values are different from those in the previous version.			Note the singular and plural in the sentence.			land11030363_makarova	0
+My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			land11030363_makarova	0
+However, there are some issues that need to be resolved before this manuscript can be considered for publication.			Generally, soil spectral measurement is carried out in a dark room to avoid interference from external light sources.			land11030363_makarova	0
+Reference to formal analytical methods must be made in a scientific manner.			Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_makarova	0
+It would be wise to give more international flavor in the sections of Introduction and Discussion.			Should the writing of the results and discussion sections be separated?			land11030363_makarova	0
+6) Discussion with new 3 subheading was significantly expanded and linked to the references, and the novelty was highlighted.			Line 264: “…, and hence parent material.” It’s not clear here.			land11030363_makarova	0
+However, there are some important issues that need to be addressed: The novelty of the study should be clearly stated in the Introduction section after the objectives are formulated.			Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_makarova	0
+is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			Line 264: “…, and hence parent material.” It’s not clear here.			land11030363_makarova	0
+Line 233: It’s usually written as “Results and discussion”.			Response to Reviewer ≠1 comment Many thanks for your response.			land11030363_makarova	0
+Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			land11030363_makarova	0
+Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			Lines 490-491: “Unlikely, a weakly represented VIP peak centered at ~850 nm was a significant predictor for the Cd content.” The meaning of this sentence is not clear, and it is a little strange.			land11030363_makarova	0
+In other words, the manuscript feels like a lot of work has been done, but it doesn't specify what the important conclusions and innovations are.			The introduction part can highlight the key points and innovations of this paper.			land11030363_makarova	0
+Please pay attention to the singular and plural in the sentence.			Readers may be more interested in your analysis and interpretation.			land11030363_makarova	0
+Please pay attention to the singular and plural in the sentence.			In addition, it needs support with more and more up-to-date recent articles.			land11030363_makarova	0
+Readers may be more interested in your analysis and interpretation.			Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+Some specific comments: The abstract could be more concise.			It undoubtedly increases your experimental workload, and you do not explain it later.			land11030363_makarova	0
+What does the controlled laboratory environment you describe look like?			However, there are some issues that need to be resolved before this manuscript can be considered for publication.			land11030363_makarova	0
+There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?			It is not clear how the 12 spectra were obtained here.			land11030363_makarova	0
+It is recommended that the number of digits after the decimal point for the value of the correlation coefficient throughout the text be consistent.			Attention should be paid to the number of decimal places in all numbers throughout the text.			land11030363_makarova	0
+Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			land11030363_makarova	0
+Reference to formal analytical methods must be made in a scientific manner.			It is also not clear how 50 spectra were obtained.			land11030363_makarova	0
+However, there are some important issues that need to be addressed:			is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			land11030363_makarova	0
+It contains new Table 2, Supplementary Table 1 and Supplementary Figure 1.			Note the singular and plural in the sentence.			land11030363_makarova	0
+Reference to formal analytical methods must be made in a scientific manner.			Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			land11030363_makarova	0
+It would be wise to give more international flavor in the sections of Introduction and Discussion.			Modified content should be marked in the revised manuscript for easy review by reviewers.			land11030363_makarova	0
+is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			land11030363_makarova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.			land11030363_makarova	0
+In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).			The conclusion is too long and does not highlight the key results.			land11030363_makarova	0
+The definite article “the” is missing in many places in the whole article.			Line 135: “impairment water resources” Is this expression appropriate?			land11030363_makarova	0
+Some specific comments: The abstract could be more concise.			It is recommended that the number of digits after the decimal point for the value of the correlation coefficient throughout the text be consistent.			land11030363_makarova	0
+However, there are some important issues that need to be addressed:			Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.			land11030363_makarova	0
+Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+prediction mechanism, prediction accuracy, etc.)			The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			land11030363_makarova	0
+My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_makarova	0
+Line 233: It’s usually written as “Results and discussion”.			Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			land11030363_makarova	0
+It is recommended that the number of digits after the decimal point for the value of the correlation coefficient throughout the text be consistent.			Line 243: “(pH, temperature, precipitation)” should not be placed after the word elevation.			land11030363_makarova	0
+Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			land11030363_makarova	0
+Line 347: “different pattern” should be “different patterns”.			My major concerns: This manuscript attempts to integrate different preprocessing methods (8 kinds), sample grouping designs (88/26, 75%/25%), and soil spectral data to predict 15 soil properties by building PLSR models.			land11030363_makarova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			Author Response Please see the attachment Author Response File: Author Response.docx			land11030363_makarova	0
+Should the writing of the results and discussion sections be separated?			What does the controlled laboratory environment you describe look like?			land11030363_makarova	0
+Table 1: The pH values are different from those in the previous version.			is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			land11030363_makarova	0
+In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).			Although this manuscript is about the application of proximal remote sensing technology to the prediction of soil properties in mountainous areas, the research focus of this manuscript is not prominent, and the conclusions drawn are not very clear.			land11030363_perova	0
+Modified content should be marked in the revised manuscript for easy review by reviewers.			Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit.			land11030363_perova	0
+Lines 456-457: What does this sentence do here?			(Line: 244-249) Line 174: “…resulting in 12 spectra”.			land11030363_perova	0
+Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			The introduction part can highlight the key points and innovations of this paper.			land11030363_perova	0
+The manuscript entitled “Determination of Mehlich 3 extractable elements with visible and near infrared spectroscopy in a mountainous agricultural land, the Caucasus Mountains” aims to evaluate the potential of VIS-NIR spectroscopy and PLSR modeling to predict M3 extractable elements (Ca, Mg, Cd, Cu, Fe, K, P, Pb, Mn, and Zn) in a representative test area of Caucasus Mountains.			Line 135: “impairment water resources” Is this expression appropriate?			land11030363_perova	0
+The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			Due to the separating the Results and Discussion section, overlapping of some comments made by reviewers and associated changes, all the text of the manuscript was modified.			land11030363_perova	0
+Line 135: “impairment water resources” Is this expression appropriate?			However, there are some important issues that need to be addressed:			land11030363_perova	0
+Line 264: “…CaCO3 and Fe),” Where is the left parenthesis?			In addition, it needs support with more and more up-to-date recent articles.			land11030363_perova	0
+It is not clear how the 12 spectra were obtained here.			During the discussion, the reasons behind the experimental results should be analyzed, not only from the correlation between the sample data but also cannot be explained by relying too much on the VIP score curve.			land11030363_perova	0
+Due to the separating the Results and Discussion section, overlapping of some comments made by reviewers and associated changes, all the text of the manuscript was modified.			Line;252-259 Line 176: “…resulted 50 spectra…”.			land11030363_perova	0
+Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			land11030363_perova	0
+Readers may be more interested in your analysis and interpretation.			It is also not clear how 50 spectra were obtained.			land11030363_perova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research, https://doi.org/10.1007/s11356-021-14673-0 Response: Thanks.			land11030363_perova	0
+In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).			(Line: 244-249) Line 174: “…resulting in 12 spectra”.			land11030363_perova	0
+The definite article “the” is missing in many places in the whole article.			Modified content should be marked in the revised manuscript for easy review by reviewers.			land11030363_perova	0
+There are 114 soil samples in this study, what is the basis for grouping by 88/26 (77.2%/22.8%)?			Table 1: The pH values are different from those in the previous version.			land11030363_perova	0
+The writing language used must be different from the way an analyst mentions it in the lab.			Modified content should be marked in the revised manuscript for easy review by reviewers.			land11030363_perova	0
+Note the singular and plural in the sentence.			is not in-depth, and the analysis of model accuracy is rarely involved in the discussion.			land11030363_perova	0
+"“Materials & Method"" section: a) There is not adequate information regarding quality control of the chemical analyses."			Line 347: “different pattern” should be “different patterns”.			land11030363_perova	0
+Especially in the results and discussion section, most of the content is to state the experimental results (correlation of data), while the discussion (eg.			Lines 145-146: “, soil reaction (pH) in 1:1 soil to KCl solution” is not clearly expressed.			land11030363_perova	0
+"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			As far as I know, most articles use only one moving window size.			land11030363_perova	0
+Please pay attention to the singular and plural in the sentence.			Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			land11030363_perova	0
+7) Conclusions was modified to show its practical outcomes for the local and international readers.			Line 233: It’s usually written as “Results and discussion”.			land11030363_perova	0
+Too many parentheses are used in the essay, which is not recommended in writing.			(Line: 244-249) Line 174: “…resulting in 12 spectra”.			land11030363_perova	0
+In addition, partial correlation analysis (e.g., Cheng et al., 2019) could be helpful and bring more insights given the fact that soil spectra are simultaneously influenced by several soil properties (e.g., SOM and Iron oxide).			"Author Response ""Please see the attachment"" Author Response File: Author Response.docx"			land11030363_perova	0
+c) Lines 125-135: There should be bibliographic references for the specific methods and their official name should be mentioned.			However, there are some issues that need to be resolved before this manuscript can be considered for publication.			land11030363_perova	0
+Line 347: “different pattern” should be “different patterns”.			Line;252-259 Line 176: “…resulted 50 spectra…”.			land11030363_perova	0
+(Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?			Some specific comments: The abstract could be more concise.			land11030363_perova	0
+Is the number of samples sufficient for the purpose of the research?			Response to Reviewer 2 comment On behalf of the co-authors, I am thankful for your valuable comments which helped us to improve the quality of this study.			land11030363_perova	0
+The introduction part can highlight the key points and innovations of this paper.			However, there are some issues that need to be resolved before this manuscript can be considered for publication.			land11030363_perova	0
+It undoubtedly increases your experimental workload, and you do not explain it later.			Line 264: “…CaCO3 and Fe),” Where is the left parenthesis?			land11030363_perova	0
+Due to the separating the Results and Discussion section, overlapping of some comments made by reviewers and associated changes, all the text of the manuscript was modified.			The spectral absorption characteristics near 1400, 1900, and 2200 nm have their corresponding influencing factors.			land11030363_perova	0
+Table 1: The pH values are different from those in the previous version.			Some specific comments: The abstract could be more concise.			land11030363_perova	0
+Line 233: It’s usually written as “Results and discussion”.			Author Response Please see the attachment Author Response File: Author Response.docx			land11030363_perova	0
+Response to Reviewer 1 comment Comments and Suggestions for Authors The manuscript entitled “Determination of Mehlich 3 extractable elements with visible 2 and near-infrared spectroscopy in a mountainous agricultural 3 land, the Caucasus Mountains” is scientifically interesting and original.			Modified content should be marked in the revised manuscript for easy review by reviewers.			land11030363_perova	0
+Table 1: The pH values are different from those in the previous version.			The introduction part can highlight the key points and innovations of this paper.			land11030363_perova	0
+Line 264: “…CaCO3 and Fe),” Where is the left parenthesis?			I suggest the following: Soil parameters affecting the levels of potentially harmful metals in Thessaly area, Greece: a robust quadratic regression approach of soil pollution prediction, Environmental Science and Pollution Research, https://doi.org/10.1007/s11356-021-14673-0 Response: Thanks.			land11030363_perova	0
+Table 1: The pH values are different from those in the previous version.			Line 191, 365: “…with different gaps and segment sizes.” “…with gap segment size of…”.			land11030363_perova	0
+7) Conclusions was modified to show its practical outcomes for the local and international readers.			Line 909: “c, f, f, a” should be superscripts, indicating the author's work unit.			land11030363_perova	0
+Lines 353-354, 374: “Savitzky-Golay 1st derivative…” “Savitzky-Golay 2nd derivatives” This is a very confusing way to write it.			(Line: 206-207) Lines 163-164: “…were conducted under a controlled laboratory environment.” Can you explain this sentence?			land11030363_perova	0
+Conclusions of the study was modified to show its practical outcomes for the local and international readers The “Introduction section” should be reworked, highlighting the relevance of this topic worldwide.			Line 347: “different pattern” should be “different patterns”.			land11030363_perova	0
+Some specific comments: The abstract could be more concise.			Lines 281, 249: Why did you use Spearman's Rho correlation coefficients earlier and Pearson correlation coefficients here?			land11030363_perova	0
+The definite article “the” is missing in many places in the whole article.			Authors should report the detection limits and the recovery of the analytic methods used, along with and the standard solutions for the calibration of the instruments.			land11030363_perova	0
+***************************************************************************** Reviewer 1 (R1):			However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310).			life12060901_makarova	0
+Authors’ response: Thank you for pointing this out.			Authors’ response: We have revised the sentence; see section 5.			life12060901_makarova	0
+The authors of this paper examined whether sociodemographic as well as health-related differences between COVID-19 patients with and without long COVID could be observed.			(2) The next major issue targets the interpretation of interaction effects.			life12060901_makarova	0
+I know, there were 7 missing values within the 310.			The authors did a great job, improved their manuscript according to my suggestions, and responded to all my questions adequately.			life12060901_makarova	0
+Hence, I urge the authors not to mislead readers and to re-write these sections.			The authors did a great job, improved their manuscript according to my suggestions, and responded to all my questions adequately.			life12060901_makarova	0
+People who have infected with COVID-19 sometimes report long lasting COVID symptoms like the loss of taste and smell.			They found no sociodemographic differences between both groups (i.e., long COVID vs. no long COVID) but, however, observed differences on three health-related outcomes (psychological distress, fatigue, and perceived stress).			life12060901_makarova	0
+The authors admit that, line 367, younger people were overrepresented, but to me, it was that older persons, especially over age 70, who were somehow missed in the survey sampling process.			Author Response We thank the reviewers for their guidance on this manuscript.			life12060901_makarova	0
+2) Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”).			The authors did a great job, improved their manuscript according to my suggestions, and responded to all my questions adequately.			life12060901_makarova	0
+In addition, the authors introduced an interaction effect between ´long COVID (yes/no) and gener (men/women) and found that men reported significantly higher psychological distress and higher fatigue than women but not for perceived stress.			***************************************************************************** Reviewer 1 (R1):			life12060901_makarova	0
+The authors consider a number of factors associated with long COVID and psychological symptoms.			Long COVID breaks that pattern - you get sick and stay sick.			life12060901_makarova	0
+However, this table is still wrong, it must be 310 and not 303.			Please use a consistent writing style throughout the whole manuscript.			life12060901_makarova	0
+Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05).			Please use a consistent writing style throughout the whole manuscript.			life12060901_makarova	0
+They found no sociodemographic differences between both groups (i.e., long COVID vs. no long COVID) but, however, observed differences on three health-related outcomes (psychological distress, fatigue, and perceived stress).			In addition, the authors introduced an interaction effect between ´long COVID (yes/no) and gener (men/women) and found that men reported significantly higher psychological distress and higher fatigue than women but not for perceived stress.			life12060901_makarova	0
+People would get really irritated and discouraged by such a situation.			**************************************************************************** Reviewer 3 (R3): The reviewed paper deals with the COVID pandemic, especially with the “long COVID” phenomenon.			life12060901_makarova	0
+However, this table is still wrong, it must be 310 and not 303.			However, this table is still wrong, it must be 310 and not 303.			life12060901_makarova	0
+However, the single numbers do not sum up to 303 but rather to 310 (13+74+220+3=310).			Hence, I urge the authors not to mislead readers and to re-write these sections.			life12060901_makarova	0
+Honestly, there is no empirical evidence that there is an association between female gender and long COVID, there was no significant difference observed between men and women (p=0.05).			Please repeat this procedure for “PSS-10” (see page 4, line 185), i.e.			life12060901_makarova	0
+R3: Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold.			However, what I disliked was the interpretation and discussion of the empirical results.			life12060901_perova	0
+Authors’ response: Thank you for the kind words.			We have corrected the manuscript according to this guidance.			life12060901_perova	0
+R3: However, I see several issues with the ending of this paper.			Psychological distress is the latent construct which is measured with the instrument GHQ.			life12060901_perova	0
+The authors did a cross-sectional online survey with a total sample of 1649 adults across four countries (Norway, UK, USA, and Australia).			Frankly speaking, the authors are overselling their results.			life12060901_perova	0
+Author Response We thank the reviewers for their guidance on this manuscript.			All comments have been addressed below, and all changes in the revised manuscript have been performed using track changes.			life12060901_perova	0
+However, I see several issues with the ending of this paper.I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential.			I would suggest another factor - frustration with not getting well.			life12060901_perova	0
+However, what I disliked was the interpretation and discussion of the empirical results.			R3: Besides these two major issues, I detected some smaller errors and miss-spellings (note, this list in not complete).			life12060901_perova	0
+They analyzed the collected data descriptively and with logistic regression models and multivariate analyses of variances (MANOVAs).			For example, see Table 4 (page 7, line 269): Column Fatgigue, why is ES not printed in bold?			life12060901_perova	0
+COVID-patients with long COVID felt higher psychological distress, higer fatigue, and higer perceived stress than COVID-patients without long COVID.			2) Table 1 (page 3, line 137): I detected an error in the third column (“COVID-19 infection”).			life12060901_perova	0
+4) Table 3, Table 4, and Table 5: The authors use a very inconsistent style of bold.			However, the authors wrote “While participants with long COVID generally perceived more psychological distress, fatigue, and stress than those without long COVID, differences were larger for men than for women” page 9, line 303), Authors’ response: Thank you for pointing this out, we have revised the sentence (see section 4.1).			life12060901_perova	0
+However, this table is still wrong, it must be 310 and not 303.			Hence, I urge the authors to re-calculate percentages in Table 1.			life12060901_perova	0
+Frankly speaking, the authors are overselling their results.			I liked the ideas and the methodological setting (i.e., online survey) of this study and I think the reported data have potential.			life12060901_perova	0
+I know, there were 7 missing values within the 310.			I found two major issues in the discussion of this paper: Authors response: Please see our comments to each of the listed issues below.			life12060901_perova	0
+The percentage of 87 long COVID (28.7%) then refers to 303 (87/303=28.7%).			Authors’ response: We have revised the sentence; see section 5.			life12060901_perova	0
+The introduction guides the reader smoothly into this topic, all necessary background information (i.e., citing appropriate literature) is given.			Frankly speaking, the authors are overselling their results.			life12060901_perova	0
+They also make quantitative measurements to describe the behaviour of the model trichomes.			Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_makarova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			life4030433_makarova	0
+The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display.			The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_makarova	0
+Round 2: Author Response to Reviewer 1 We have addressed the reviwer’s requests: (1)			I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_makarova	0
+The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_makarova	0
+While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			I have a few comments, suggestions, and questions.			life4030433_makarova	0
+They also make quantitative measurements to describe the behaviour of the model trichomes.			In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			life4030433_makarova	0
+The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			life4030433_makarova	0
+Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_makarova	0
+The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			In response to the reviewers’ comments: (1) We initially attempted to use the bending modulus implied from Boal and Ng’s study, however we found that during simulations this value seemed to be too low.			life4030433_makarova	0
+While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			Qualitative comparisons are made to the fossil record and modern experiments.			life4030433_makarova	0
+Round 2: Author Response to Reviewer 1 We have addressed the reviwer’s requests: (1)			I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_makarova	0
+The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			The paper can be published once the authors more clearly confront their disagreement with the results of Boal and Ng.			life4030433_makarova	0
+The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			I have a few comments, suggestions, and questions.			life4030433_makarova	0
+Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_makarova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			life4030433_makarova	0
+I have a few comments, suggestions, and questions.			The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display.			life4030433_makarova	0
+Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			life4030433_makarova	0
+Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			life4030433_makarova	0
+The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			They also explore behaviour in much of the available parameter space.			life4030433_makarova	0
+I have a few comments, suggestions, and questions.			I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_makarova	0
+Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			The trichomes were completely flaccid and would collapse into a heap as soon as one collided with another trichome, unlike the smooth sinuous shape filamentous cyanobacteria often display.			life4030433_makarova	0
+I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_makarova	0
+Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_makarova	0
+As a result, I do not recommend that this paper should be published in its current form.			They also explore behaviour in much of the available parameter space.			life4030433_perova	0
+Simulations using the model address the observation of reticulate patterns in experiments with filamentous cyanobacteria.			The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			life4030433_perova	0
+As a result, I do not recommend that this paper should be published in its current form.			Thus, the length of a filament is not a model parameter.			life4030433_perova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+).			life4030433_perova	0
+They also explore behaviour in much of the available parameter space.			However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			life4030433_perova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			Rather, length is determined dynamically as ~ N k τ + (10)			life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			life4030433_perova	0
+I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			life4030433_perova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			life4030433_perova	0
+As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+).			While the authors might be able to find a similar phenomenological agreement with more reasonable parameters, it will change all of their results and amounts to a substantial revision.			life4030433_perova	0
+Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			Second Round of Evaluation Round 2: With the exception of their response to one key point, the authors' responses are satisfactory.			life4030433_perova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			Rather, length is determined dynamically as ~ N k τ + (10)			life4030433_perova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			I do not feel that any of these points demand major changes to the manuscript, which is generally fine in the present form.			life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			However, their parameterization of the bending modulus is not acceptable because it leads to an unreasonably large (0.5 metre) persistence length for a bacterial trichome.			life4030433_perova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			Thus, the length of a filament is not a model parameter.			life4030433_perova	0
+In their response, the authors mention that with the lower value from Boal and Ng the trichomes collapse into a heap, and so a larger bending modulus was necessary.			The model is successful in producing local alignment of filaments and some sharp bends—both characteristics of the experimental system.			life4030433_perova	0
+As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+).			Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_perova	0
+They also argue that a very large persistence length implies that the cyanobacteria are stiff enough to largely be unaffected by thermal fluctuations.			Their suggested bending modulus value implies a very high persistence length of 0.5 metres.			life4030433_perova	0
+Qualitative comparisons are made to the fossil record and modern experiments.			The authors find that for a certain parameter regime, they observe in their simulations patterns with characteristics similar to the characteristics of the experimental patterns.			life4030433_perova	0
+They also make quantitative measurements to describe the behaviour of the model trichomes.			They also explore behaviour in much of the available parameter space.			life4030433_perova	0
+As I recall, these cyanobacteria reproduce (with doubling time τ ) by adding cells to the filament and that filaments periodically break (rate k+).			Thus, the length of a filament is not a model parameter.			life4030433_perova	0
+Thus, the length of a filament is not a model parameter.			Round 1: Author Response to Reviewer 2 We would like to thank the reviewer for his/her thorough review of our manuscript and thoughtful comments.			life4030433_perova	0
+Round 1: Author Response to Reviewer 1 We would like to thank the reviewer for taking the time to do a thorough review of our paper.			As a result, I do not recommend that this paper should be published in its current form.			life4030433_perova	0
+Notably, the introduction of reproduction does not introduce another parameter.			Round 1: The authors develop and analyze a model for the dynamics of filamentous bacteria gliding in two dimensions.			life4030433_perova	0
+Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			life4040819_makarova	0
+Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			life4040819_makarova	0
+R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			life4040819_makarova	0
+Within the text, corrected words are highlighted.			By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			life4040819_makarova	0
+A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			In addition, Figure 3A was replaced by a figure of better contrast.			life4040819_makarova	0
+Using TEM, the authors observed junctional pore complexes arrayed at septa, and extending through the cell envelope to the exterior of the cell, and potentially identified basal body complexes at the cytoplasmic face of the inner membrane.			presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			life4040819_makarova	0
+R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			life4040819_makarova	0
+However, a number of shortcomings need to be addressed before the manuscript could be accepted for publication.			The observations and models are consistent with previously published work on motility of various cyanobacteria and provide new insights into the mechanics of motility of cyanobacteria that are interesting.			life4040819_makarova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			Response: We thank the reviewers for giving us helpful comments.			life4040819_makarova	0
+A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			life4040819_makarova	0
+Within the text, corrected words are highlighted.			The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			life4040819_makarova	0
+R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			life4040819_makarova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			In addition, Figure 3A was replaced by a figure of better contrast.			life4040819_makarova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			life4040819_makarova	0
+The contribution of motility to the formation of supracellular structure is generally an area of research regarding cyanobacteria that has been overlooked.			The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			life4040819_makarova	0
+Within the text, corrected words are highlighted.			This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			life4040819_makarova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			life4040819_makarova	0
+This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			Within the text, corrected words are highlighted.			life4040819_makarova	0
+Response: We thank the reviewers for giving us helpful comments.			The text was revised according to the comments.			life4040819_makarova	0
+A few major comments can be found below concerning R2 the possible presence of type IV pili in their EM images, some confusion regarding descriptions of motility analysis and others.			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_makarova	0
+The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			life4040819_makarova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_makarova	0
+The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			Response: We thank the reviewers for giving us helpful comments.			life4040819_perova	0
+In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			Finally, the authors performed a detailed characterization of the motility of individual filaments of the organism by time lapse microscopy providing additional evidence that would appear to support polysaccharide secretion as the driving force for motility, and provide a model for how this motility drives the formation of supercellular structures; in this case, spirals.			life4040819_perova	0
+presents characterization of the growth dynamics and motility of a newly isolated Phormodium strain.			Round 1: and Author Response The manuscript “Cellular dynamics drives emergence of supracellular structure in the cyanobacterium Phormidium sp.			life4040819_perova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			A model is presented to describe how the rotation of the filament causes left-handed sliding for the portion of the filament not encased in a slime sheath, which leads to formation of counter-clockwise spirals, the predominant macroscopic feature of colonies noted.			life4040819_perova	0
+Within the text, corrected words are highlighted.			Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			life4040819_perova	0
+Within the text, corrected words are highlighted.			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Cellular Dynamics Drives Emergence of Supracellular Structure in the Cyanobacterium Phormidium sp.			life4040819_perova	0
+In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			life4040819_perova	0
+By characterizing motility of colonies, as well as single-filament motility of short filaments, the authors find that increasing the agar concentration of the media results in smaller colonies, and slower individual filament movement.			R4 Because the EM figures are difficult to view within the Word document, we provide a high-quality figure as a supplement.			life4040819_perova	0
+Round 1: and Author Response The manuscript “Cellular dynamics drives emergence of supracellular structure in the cyanobacterium Phormidium sp.			Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			life4040819_perova	0
+Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			In particular, the appearance of bulges in the extracellular sheath adjacent to the site of the junctional pores appears to be consistent with this mode of motility.			life4040819_perova	0
+Within the text, corrected words are highlighted.			The text was revised according to the comments.			life4040819_perova	0
+Response: We thank the reviewers for giving us helpful comments.			Using TEM, the authors observed junctional pore complexes arrayed at septa, and extending through the cell envelope to the exterior of the cell, and potentially identified basal body complexes at the cytoplasmic face of the inner membrane.			life4040819_perova	0
+This study is our new exploration into a new field, and the comments from different fields help us to revise our manuscript.			In addition, Figure 3A was replaced by a figure of better contrast.			life4040819_perova	0
+Additionally, the authors sequenced the genome of this organism and identified homologs to genes conserved among filamentous cyanobacteria that have been demonstrated to be essential for motility in Nostoc punctiforme and may encode for the junctional pore proteins (hps locus) or a chemotaxis system (hmp locus).			Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			life4040819_perova	0
+R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			In addition, Figure 3A was replaced by a figure of better contrast.			life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			life4040819_perova	0
+The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			life4040819_perova	0
+Overall, this study presents observations related to slime-mediated motility on multiple scales (molecular, single-filament and colony), and integrates these findings into a coherent model for the relationship between the cellular organization and the behavior of the colony.			R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			life4040819_perova	0
+Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			R3 Minor comments The addition of a time stamp to the supplemental movies would be useful 			life4040819_perova	0
+Additionally, individual filaments were found to undergo polarity reversals, and the maximum velocity was observed immediately following the reversal.			The authors demonstrate the presence of putative junctional pores by electron microscopy and other ultrastructural data that would appear to support polysaccharide secretion as the driving force for motility in this organism.			life4040819_perova	0
+The observations and models are consistent with previously published work on motility of various cyanobacteria and provide new insights into the mechanics of motility of cyanobacteria that are interesting.			Response: We thank the reviewers for giving us helpful comments.			life4040819_perova	0
+The authors’ conclusions are generally well supported and the electron microscopy provides some of the clearest images to date of the junctional pores.			Within the text, corrected words are highlighted.			life4040819_perova	0
+The alternative views he previously generated are much expanded in this manuscript.			I must note that a careful evaluation of all facets requires expertise in a multitude of disciplines (from prebiotic chemistry and structural biology to evolutionary bioinformatics and biochemistry) and considerable time, none of which I possess.			life4041050_makarova	0
+I have only a few comments, which might improve the manuscript.			He reviews some of his previous work and presents an alternative to the dominant ‘Ancient RNA world’ hypothesis.			life4041050_makarova	0
+In light of recent research developments and argumentation (some of it reviewed), his views should be considered a welcome addition to the many ideas that populate the “origin of life” field of inquiry that counter the dominant paradigm.			I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			life4041050_makarova	0
+Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			life4041050_makarova	0
+I have only a few comments, which might improve the manuscript.			The article demonstrates that the iron-sulfur surface metabolism theory at high temperatures has a much better explanatory power than the RNA World theory.			life4041050_makarova	0
+The article demonstrates that the iron-sulfur surface metabolism theory at high temperatures has a much better explanatory power than the RNA World theory.			I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			life4041050_makarova	0
+Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			The alternative views he previously generated are much expanded in this manuscript.			life4041050_makarova	0
+The alternative views he previously generated are much expanded in this manuscript.			The article demonstrates that the iron-sulfur surface metabolism theory at high temperatures has a much better explanatory power than the RNA World theory.			life4041050_makarova	0
+Round 1: and Author Response The manuscript comprises an exciting account on the origin of life with emphasis on the emergence and function of RNA.			Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			life4041050_makarova	0
+Round 1: and Author Response The manuscript comprises an exciting account on the origin of life with emphasis on the emergence and function of RNA.			I have only a few comments, which might improve the manuscript.			life4041050_makarova	0
+The alternative views he previously generated are much expanded in this manuscript.			I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			life4041050_makarova	0
+The article demonstrates that the iron-sulfur surface metabolism theory at high temperatures has a much better explanatory power than the RNA World theory.			I have however a number of quibbles that if addressed could increase the accuracy, value and impact of the manuscript.			life4041050_makarova	0
+Therefore, my comments will be slanted by my own expertise and will only serve the author as a partial devil’s advocate effort General commentary			In light of recent research developments and argumentation (some of it reviewed), his views should be considered a welcome addition to the many ideas that populate the “origin of life” field of inquiry that counter the dominant paradigm.			life4041050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+We thus only briefly discuss names and taxonomy in our analysis.			Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			life5010050_makarova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_makarova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			We thus only briefly discuss names and taxonomy in our analysis.			life5010050_makarova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_makarova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			life5010050_makarova	0
+Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_makarova	0
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			Given that both reviewers commented on this issue, we tried to clarify our aims and included a sentence at the end of the introduction, in order to clarify the point.			life5010050_makarova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			life5010050_makarova	0
+ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_makarova	0
+We thus only briefly discuss names and taxonomy in our analysis.			We thus only briefly discuss names and taxonomy in our analysis.			life5010050_makarova	0
+Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			More citations of modern literature would be expectable and useful.			life5010050_perova	0
+However, the only one universal method is quite insufficient in microorganisms, and the combination of all possible classification criteria is desirable.			More citations of modern literature would be expectable and useful.			life5010050_perova	0
+My critics should inform authors about more complicated problematics of this whole group, and several principles should be respected and accepted for future cyanobacterial research.			It is overall well-written and innovative work, but there are few things to consider in methods and results, which should be revised, and some formal flaws.			life5010050_perova	0
+Second Round of Evaluation Round 2: I have had the more critical remarks to the manuscript; however, I think that the presented data are useful and should be published.			Round 2: I accept the paper in the present form.			life5010050_perova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			ISSN 2075-1729 www.mdpi.com/journal/life Peer-Review Record: Does a Barcoding Gap Exist in Prokaryotes?			life5010050_perova	0
+Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			We thus only briefly discuss names and taxonomy in our analysis.			life5010050_perova	0
+Second Round of Evaluation Round 2: I have had the more critical remarks to the manuscript; however, I think that the presented data are useful and should be published.			We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			life5010050_perova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			My critics should inform authors about more complicated problematics of this whole group, and several principles should be respected and accepted for future cyanobacterial research.			life5010050_perova	0
+We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			Round 1: and Author Response The authors tested novel approach to identification of species in cyanobacteria based on 16S rRNA based on identification of barcoding gaps previously used in animals.			life5010050_perova	0
+The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			However, the only one universal method is quite insufficient in microorganisms, and the combination of all possible classification criteria is desirable.			life5010050_perova	0
+My critics should inform authors about more complicated problematics of this whole group, and several principles should be respected and accepted for future cyanobacterial research.			We do not deal with taxonomy at all, only with testing the presence of a barcoding gap.			life5010050_perova	0
+We thus only briefly discuss names and taxonomy in our analysis.			Summary: The manuscript is surely useful and, in principle, it presents a good contribution to the recent methodological trends (polyphasic approach) in cyanobacterial taxonomy, this conclusion should be the main result from this work.			life5010050_perova	0
+Second Round of Evaluation Round 2: I have had the more critical remarks to the manuscript; however, I think that the presented data are useful and should be published.			To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			life5010050_perova	0
+The molecular methods must be evidently preferred in this work, but other approaches (biochemical, ecophysiological, ecological, morphological) must be included into the final evaluation.			Of course, the complex of all these methods must be corrected with the progress of science, and especially for cyanobacteria is necessary to apply the so called “polyphasic approach” for their classification.			life5010050_perova	0
+To do so, we did not download named sequences from GenBank, but used a carefully annotated and checked database with a built-in phylogeny that would allow us to obtain monophyletic clades, regardless of their names.			We thus only briefly discuss names and taxonomy in our analysis.			life5010050_perova	0
+More citations of modern literature would be expectable and useful.			I believe that the article with proposed changes will be useful for future research.			life5010050_perova	0
+We hope that now both reviewers, and all other potential readers, will not be misled by our approach.			Thus this study intends only to verify the existence of a barcoding gaps within the 16S rRNA sequences of certain cyanobacterial groups, and by no means revise or confirm the complex cyanobacterial taxonomy.”			life5010050_perova	0
+Round 1: and Author Response The article “Emergent Chemical Behaviour in Variable-Volume Protocells” focuses on the consequences that a changing volume may have on a set of reactions encapsulated within a semipermeable vesicle; in particular, this kind of compartment can degenerate existing bistable reactions, or promote emergent bistability from very simple reactions, which are not bistable in bulk conditions.			The mathematical methods used to deal with the problem are based on approximations and require the support of a huge number of simulations; on the other side, the approximations themselves constitute an interesting feature of the article and allow interesting discussions on the involved physical situations.			life5010181_perova	0
+Produces a dynamic model for chemical evolution rather than traditional deterministic, static, systems that requires centralised information to direct evolution, such as a constant gradient.			Also a very thorough understanding of the limitations of the system are presented as well as proposals for even more complex instabilties that may underpin lifelike events.			life5010181_perova	0
+The authors discuss these issues by means of differential equations and simulations.			Very well presented, clear and graphically illustrated experimental findings that make a clear case for the model.			life5010181_perova	0
+The mathematical methods used to deal with the problem are based on approximations and require the support of a huge number of simulations; on the other side, the approximations themselves constitute an interesting feature of the article and allow interesting discussions on the involved physical situations.			Remarkably, even reaction systems that have no chemical species in common could become R2 indirectly coupled to each other through the volume they share, by means of a sort of osmotic coupling.			life5010181_perova	0
+Round 1: and Author Response The article “Emergent Chemical Behaviour in Variable-Volume Protocells” focuses on the consequences that a changing volume may have on a set of reactions encapsulated within a semipermeable vesicle; in particular, this kind of compartment can degenerate existing bistable reactions, or promote emergent bistability from very simple reactions, which are not bistable in bulk conditions.			Very well presented, clear and graphically illustrated experimental findings that make a clear case for the model.			life5010181_perova	0
+The authors discuss these issues by means of differential equations and simulations.			As the authors acknowledge, the article is the first step toward a more complete understanding of an interesting class of possible chemo-physical phenomena.			life5010181_perova	0
+Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			Second Round of Evaluation Round 2: and Author Response			life5010403_makarova	0
+The data presented is robust and worthy of publication but should be discussed more critically with regard to published literature (outlined below) I suggest some minor suggestions that should be incorporated before publications.			Second Round of Evaluation Round 2: and Author Response			life5010403_makarova	0
+Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			This is done for a number of globally significance marine microbes of Syn and Pro lineage.			life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			life5010403_makarova	0
+The data presented is robust and worthy of publication but should be discussed more critically with regard to published literature (outlined below) I suggest some minor suggestions that should be incorporated before publications.			Round 1: and Author Response The authors present a dataset where they have characterised the relative abundance of components of the photosynthetic electron transfer chain and related this to the measured rate of electron transfer from PSII.			life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			The approach and methods are robust and the ability to quantify components of the photosynthetic electron transfer chain is powerful.			life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			This is done for a number of globally significance marine microbes of Syn and Pro lineage.			life5010403_makarova	0
+Second Round of Evaluation Round 2: and Author Response			The data presented is robust and worthy of publication but should be discussed more critically with regard to published literature (outlined below) I suggest some minor suggestions that should be incorporated before publications.			life5010403_makarova	0
+This is done for a number of globally significance marine microbes of Syn and Pro lineage.			provides us with estimates of the concentrations of various proteins representing the PSII, PSI, cytb6 and Rubisco in there different cyanobacteria.			life5010403_makarova	0
+entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			life5010432_makarova	0
+entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			Functional annotation of genes through protein expression is highly needed in our time of exponentially growing sequence data.			life5010432_makarova	0
+Round 1: and Author Response I found this research interesting, relevant and novel.			CC9605 is an open ocean strain and does not encounter high nitrite concentrations, while CC9311 is a coastal strain that deals with a range of nitrate/nitrite availability.			life5010432_makarova	0
+(3) p. 5 line 5, “abbreviation” has been changed to “abbreviations”.			R2 This article will be of interest to general microbiologists, specialists in cyanobacteria, and especially researchers that conduct environmental research and also genome annotations.			life5010432_makarova	0
+Such differences can be explained by habitat adaption of each strain.			Such differences can be explained by habitat adaption of each strain.			life5010432_makarova	0
+R2 This article will be of interest to general microbiologists, specialists in cyanobacteria, and especially researchers that conduct environmental research and also genome annotations.			The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			life5010432_makarova	0
+It was particularly interesting to see the differences in protein functionality among the strains of marine Synechococcus.			Such differences can be explained by habitat adaption of each strain.			life5010432_makarova	0
+(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			R2 This article will be of interest to general microbiologists, specialists in cyanobacteria, and especially researchers that conduct environmental research and also genome annotations.			life5010432_perova	0
+CC9605 is an open ocean strain and does not encounter high nitrite concentrations, while CC9311 is a coastal strain that deals with a range of nitrate/nitrite availability.			It was particularly interesting to see the differences in protein functionality among the strains of marine Synechococcus.			life5010432_perova	0
+Such differences can be explained by habitat adaption of each strain.			(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			life5010432_perova	0
+The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			The characterized here gene, nitM, can serve as a molecular marker in environmental studies.			life5010432_perova	0
+Functional annotation of genes through protein expression is highly needed in our time of exponentially growing sequence data.			Functional annotation of genes through protein expression is highly needed in our time of exponentially growing sequence data.			life5010432_perova	0
+R2 This article will be of interest to general microbiologists, specialists in cyanobacteria, and especially researchers that conduct environmental research and also genome annotations.			R3 Other changes (1) p. 1 line 26, “α-cyanobacteira” has been changed to “α-cyanobacteria”.			life5010432_perova	0
+(2) p. 2 lines 2–3, “α-cyanobacteiral” has been changed to “α-cyanobacterial”.			entitled “Functional Characterization of the FNT Family Nitrite Transporter of Marine Picocyanobacteria” is a very interesting piece of work since it shows the functional characterization as nitrite transporter of several nitM proteins from different picocyanobacteria, showing also fruitful comparisons of their sequences that lead to characterize the c-terminal region of a-cyanobacterial NitM proteins as an inhibitory domain of nitrite transport.			life5010432_perova	0
+Results also provide additional support to recently proposed archaeal phyla and halobacterial orders.			If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.”			life5010949_makarova	0
+About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			life5010949_makarova	0
+This kind of research should be encouraged further because old taxonomic paradigms must be systematically reviewed based on new genomic data.			Even if defined in the future, it must be lineage-dependent.			life5010949_makarova	0
+Even if defined in the future, it must be lineage-dependent.			It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			life5010949_makarova	0
+This kind of research should be encouraged further because old taxonomic paradigms must be systematically reviewed based on new genomic data.			Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			life5010949_makarova	0
+I recommend acceptance for publication in Life Sciences journal.			Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			If all genomes from a taxon appear exclusively in a tree branch, the branch is said to be monophyletic.”			life5010949_makarova	0
+present a comparative analysis of the taxonomic classification of the Archaea domain.			The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			life5010949_makarova	0
+About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			life5010949_makarova	0
+R13 Round 2: and Author Response Authors have substantially improved the article, including language corrections, and have provided extensive clarifications to all initial criticisms.			are describing a comprehensive analysis of archaeal phylogeny with a genome-based alignment free method, and then comparing the findings to 16S rRNA based phylogenies.			life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			Even if defined in the future, it must be lineage-dependent.			life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+present a comparative analysis of the taxonomic classification of the Archaea domain.			For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			life5010949_makarova	0
+Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			life5010949_makarova	0
+I recommend acceptance for publication in Life Sciences journal.			The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			life5010949_makarova	0
+Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			I recommend acceptance for publication in Life Sciences journal.			life5010949_makarova	0
+Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			Evaluation I found major concerns along this manuscript, and suggest a substantial revision.			life5010949_makarova	0
+However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			life5010949_makarova	0
+The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+For example, it cannot be expected that the same degree of dissimilarity may be used to delineate classes in all phyla.			R13 Round 2: and Author Response Authors have substantially improved the article, including language corrections, and have provided extensive clarifications to all initial criticisms.			life5010949_makarova	0
+Perhaps an additional discussion point would be the advantages of using CVtree approach with regular concatenated proteins.			present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			General Comments The topic is relevant for microbial taxonomy.			life5010949_makarova	0
+About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+I recommend acceptance for publication in Life Sciences journal.			General Comments The topic is relevant for microbial taxonomy.			life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			It has been known that tiny genomes of endosymbiont microbes often tend to move towards baseline of a tree and distort the overall picture.			life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			I recommend acceptance for publication in Life Sciences journal.			life5010949_makarova	0
+The tree was compared with several 16S rRNA trees reported in the literature, and the differences were minor.			The tree was compared with several 16S rRNA trees reported in the literature, and the differences were minor.			life5010949_makarova	0
+I recommend acceptance for publication in Life Sciences journal.			present a comparative analysis of the taxonomic classification of the Archaea domain.			life5010949_makarova	0
+However, I am personally not convinced that whole genomes bring additional advantages, or if we are better off by just using a conserved set of proteins.			Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+R13 Round 2: and Author Response Authors have substantially improved the article, including language corrections, and have provided extensive clarifications to all initial criticisms.			About 180 archaeal genomes were used to calculate a new tree topology using CVTree.			life5010949_makarova	0
+The idea to use whole genome data for archaeal phylogenies is interesting, as the 16S rRNA phylogenies can be poor in resolving relationships among archaeal lineage due to GC content bias in hyperthermophilic Archaea.			Round 1: and Author Response Summary Zuo et al.			life5010949_makarova	0
+The latter can be assigned only after comparison with a reference taxonomy which is not a rigid framework but a modifiable system.			Though a dissimilarity measure figures in the CVTree algorithm, it is not realistic to delineate taxa by using this measure at least for the time being.			life5010949_makarova	0
+Here monophyly must be understood in a pragmatic way restricted to the given set of input data and the reference taxonomy.			Even if defined in the future, it must be lineage-dependent.			life5010949_makarova	0
+It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			It is the only described representative of a newly proposed phylum Nanoarchaeota and it cuts into the otherwise monophyletic phylum Euryarchaeota.			life5010949_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			life5021101_makarova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			life5021101_makarova	0
+I also have questions and comments for acceptance.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			life5021101_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			life5021101_makarova	0
+I also have questions and comments for acceptance.			I also have questions and comments for acceptance.			life5021101_makarova	0
+I also have questions and comments for acceptance.			According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			life5021101_makarova	0
+The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			life5021101_makarova	0
+According to the authors, this is the first report for the measurement of bending elasticity modulus for archaeal lipid-based GVs.			The observed fluctuations of the membrane do not destabilize it.			life5021101_makarova	0
+Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			I also have questions and comments for acceptance.			life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			I also have questions and comments for acceptance.			life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			Stroboscopic illumination measurements of GVs and theoretical models about shape fluctuation of GVs revealed that the archaeal lipid-based GVs showed the almost same value of bending elasticity modulus with synthetic lipid-based GVs.			life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			life5021101_makarova	0
+Round 1: and Author Response This manuscript described the measurement of bending elasticity of giant vesicles (GVs) prepared by archaea-derived lipids.			The observed fluctuations of the membrane do not destabilize it.			life5021101_makarova	0
+I also have questions and comments for acceptance.			The measurement itself is novel, but same methods and theories have been reported in the authors’ previous papers.			life5021101_makarova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			The observed fluctuations of the membrane do not destabilize it.			life5021101_makarova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+Second Round of Evaluation Round 2: and Author Response It is also not clear about the “thermally induced shape fluctuations”, such as the temperatures and incubation length and etc., especially this is a hyperthermophilic archaeal membrane vesicle, which would be expected to be more thermostable than those eukaryotic ones.			The observed fluctuations of the membrane do not destabilize it.			life5021101_perova	0
+However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+The observed fluctuations of the membrane do not destabilize it.			However, the comment of the reviewer is notable and we state in the conclusion of the manuscript (Lines 229–232) that at room temperature the thermophilicity is not necessarily related to the bending elasticity.			life5021101_perova	0
+In summary, this paper is of good quality and of enormous value, and should be published.			This is a great paper to aim at such a direction.			life5021427_makarova	0
+While not especially flashy or exciting, the authors tackle a serious problem, the reliability of annotations in publically available databases.			Importance: Archaea is/are the third domain of life, no more or less!			life5021427_makarova	0
+While not especially flashy or exciting, the authors tackle a serious problem, the reliability of annotations in publically available databases.			The authors describe an alternative, manual curation strategy aimed at avoiding over-annotation.			life5021427_perova	0
+The manuscript describes in detail the approach used by the HaloLex genome annotation system to obtain reliable, consistent annotations, based on “Gold Standard Proteins”.			The curation procedure is done, to my mind, very well; and I am confident that this will help the archaea community enormously.			life5021427_perova	0
+The manuscript contains scientific material that can be published as a short communication.			Line 38: Deinococcus radiodurans should be written in italic.			life6020022_makarova	0
+First of all it is necessary to change the title and at least remove “The testing”.			The manuscript contains scientific material that can be published as a short communication.			life6020022_makarova	0
+Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.			Line 78: Please explain in the text what is “a vacuum treatment control”.			life6020022_makarova	0
+Line 38: Deinococcus radiodurans should be written in italic.			The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			life6020022_makarova	0
+Minor comments (please see changes in text at the relevant lines)			Minor comments Lines 31-32: Bacillus subtilis and B.pumilus should be written in italic.			life6020022_makarova	0
+Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?			As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV.			life6020022_makarova	0
+The exposure times varied from 1.5 to 45 min with different experiment Conditions/procedures without resulting in a significant inactivating effect on the bacterial cells.			Knowing the grain sizes and assuming a homogeneous distributrion of cells and grains or a measurement of the real distribution an assessment can be made about the shading effect of the grains with respect to UV radiation.			life6020022_makarova	0
+The manuscript contains scientific material that can be published as a short communication.			Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.			life6020022_makarova	0
+The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			However, there are some remarks on the justification of objectives of the study and presentation of the results.			life6020022_makarova	0
+However, there are some remarks on the justification of objectives of the study and presentation of the results.			As the samples are placed in an ICP discharge, ions towards the samples are only accelerated by several tenth of eV.			life6020022_makarova	0
+Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?			The manuscript contains scientific material that can be published as a short communication.			life6020022_makarova	0
+The manuscript contains scientific material that can be published as a short communication.			The mineralogical composition is given in table, but not the grain size distribution.			life6020022_makarova	0
+Estimations of the average free length of path for the inactivating plasma components might have been helpful for experiment planning.			With the knowledge of the spectral irradiance in the UV range from 100 to 400 nm the authors could have made a calculation to estimate the inactivation of D. radiodurans by the UV component of the plasma.			life6020022_makarova	0
+In addition, I think, a control consisting of D. radiodurans cells before and after sterilization is missing to determine the role of the Martian Analog soils.			For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description.			life6020022_makarova	0
+Line 78: Please explain in the text what is “a vacuum treatment control”.			The other potentially inactivating components of the plasma are not specified in the manuscript.			life6020022_perova	0
+Line 38: Deinococcus radiodurans should be written in italic.			If I understood correctly, you used only one medium.			life6020022_perova	0
+The mineralogical composition is given in table, but not the grain size distribution.			However, there are some remarks on the justification of objectives of the study and presentation of the results.			life6020022_perova	0
+The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			Reply: The names are written in italics now, the mistake must have happened during the editing of the text.			life6020022_perova	0
+Line 78: Please explain in the text what is “a vacuum treatment control”.			The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			life6020022_perova	0
+Line 78: Please explain in the text what is “a vacuum treatment control”.			Line 78: Please explain in the text what is “a vacuum treatment control”.			life6020022_perova	0
+The manuscript contains scientific material that can be published as a short communication.			However, there are some remarks on the justification of objectives of the study and presentation of the results.			life6020022_perova	0
+For the argon-oxygen gas mixture it is stated that ‘high amounts of reactive oxygen species are formed in the plasma’ again without any qualitative and quantitative description.			It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are.			life6020022_perova	0
+It is only stated that the argon-nitrogen mixture ‘mainly emits high amounts of radiation in the wavelength region from 100 nm to 400 nm’ without specifying what high fluences at the sample site are.			First of all it is necessary to change the title and at least remove “The testing”.			life6020022_perova	0
+Discussion How do you explain the difference between the results when using JSC Mars-1A analog and the other soils?			While writing D. radiodurans R1 has always necessary to note that this is type strain R1T.			life6020022_perova	0
+First of all it is necessary to change the title and at least remove “The testing”.			In the manuscript ‘Testing the Effects of Low-Temperature Plasma-Sterilization on Mars Analog Soil Samples Intermixed with Deinococcus radiodurans’ by Schirmack et al., the results of the application of low-temperature plasma on Mars regolth simulant mixed with cells of the bacterium D. radiodurans were reported.			life6020022_perova	0
+The authors’ like to thank the reviewer for the helpful comments and suggested corrections to improve the manuscript.			The authors should have estimated the maximal theoretically achievable effect and to plan the experiments accordingly.			life6020022_perova	0
+Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.			Dear academic editors I response to all the comments by reviewer 1.			life6030029_makarova	0
+I agree that the question of the relationship between RNA World and the environment is important, but there is nothing in the main part of this article that addresses this question.			It is also fairly rambling: for example, lines 95-106 could be removed, Answer:			life6030029_makarova	0
+Can we not have an RNA polymerase that uses other copies of itself as a template?			The input and output would not necessarily need to be controlled by another gene.			life6030029_makarova	0
+Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included.			Reviewer 1 line 38 - The linking of 'other molecules' to 'phenotype' here is a little odd.			life6030029_makarova	0
+The connection between the reactions catalyzed by the ribozymes and the concepts of CCSI and CMIO is not very clear at this point.			It is also fairly rambling: for example, lines 95-106 could be removed, Answer:			life6030029_makarova	0
+This section seems to mix up two important questions.			The input and output would not necessarily need to be controlled by another gene.			life6030029_makarova	0
+More fundamentally perhaps, the distinction that is made between CCSI and CMIO is, as the author himself notes, remarkably similar to that between genotype and phenotype, and information and function, and even replication and metabolism.			Fig 5 - drawing parallels between systems of different levels of complexity (from prokaryotes up to civilizations) is interesting, but it is a bit distracting at this point.			life6030029_makarova	0
+So I think this idea needs to be explained and justified in a biological context.			The abstract promised to talk about RNA World and the origin of life.			life6030029_makarova	0
+ lines 93-106 - I mostly agree with this paragraph but it is not said very clearly.			Once again in Fig 8 I think that the cycle is over-emphasized and the throughput is not included.			life6030029_makarova	0
+Another interesting idea that is also raised is the possible importance of RNA solubility at high temperatures, though it does seem that the author is battling nobly (if possibly in vain) to reconcile the RNA world hypothesis with a hydrothermal vent origin of life scenario.			Also, as far as this reviewer is aware, there are no known general (= nonspecific) RNA replicase ribozymes consisting of only 100 nucleotides.			life6030029_makarova	0
+The input and output would not necessarily need to be controlled by another gene.			Fig 7 - Viewing a metabolism as a cycle seems to be only half the story.			life6030029_makarova	0
+One example obvious to me is the reference to one of my papers [ref.			There are many quantitative traits affected by many genes.			life6030029_makarova	0
+It is also fairly rambling: for example, lines 95-106 could be removed, Answer:			Also the diagram does not indicate whether the ribozymes are made by the cycle.			life6030029_makarova	0
+Reviewer 2 According to the abstract, this manuscript covers a number of different areas relating to the RNA world hypothesis, including its compatibility with (likely) primitive Earth conditions; the likely characteristics of an RNA world; and, finally, the relationship between an early life-like RNA world system(s) and other life-like systems.			For example there must be ways of making nucleotides from scratch.			life6030029_makarova	0
+It is also fairly rambling: for example, lines 95-106 could be removed, Answer:			Author Response Author Response File: Author Response.pdf			life6030029_makarova	0
+The issue for the origin of life is how to get to a prokaryote.			Maybe there is some slightly different environment that we would still call hydrothermal in which RNA is sufficiently stable.			life6030029_perova	0
+These issues need to be considered separately.			"Statements such as in line 232: ""the first step is to limit the characteristics of life-like systems into life on Earth"" leave one scratching ones head."			life6030029_perova	0
+Was there a way of reproducibly making the same amino acid sequence without translating it from RNA?			But several questions still remain - were there long peptides before long RNA strands?			life6030029_perova	0
+The citations 52 and 53 are in fields that are not related to biology and origin of life.			This section seems to mix up two important questions.			life6030029_perova	0
+These issues need to be considered separately.			Author Response Author Response File: Author Response.pdf			life6030029_perova	0
+The second point is that the paper contains too many errors.			Author Response Author Response File: Author Response.pdf			life6030029_perova	0
+There may be a link from one gene to one protein, but one protein is not really a phenotype.			Author Response Author Response File: Author Response.pdf			life6030029_perova	0
+One example obvious to me is the reference to one of my papers [ref.			The conclusion comes back to the point about primitive Earth environment.			life6030029_perova	0
+It would seem difficult to evolve two separate ribozyme functions at the same time.			It is only when we get to paragraph 3.1 that we get to the point of the article.			life6030029_perova	0
+Another interesting idea that is also raised is the possible importance of RNA solubility at high temperatures, though it does seem that the author is battling nobly (if possibly in vain) to reconcile the RNA world hypothesis with a hydrothermal vent origin of life scenario.			Are you saying a cell interacts with the environment but a single gene does not?			life6030029_perova	0
+The issue for the origin of life is how to get to a prokaryote.			One example obvious to me is the reference to one of my papers [ref.			life6030029_perova	0
+(ii) Do we need the RNA system to be enclosed in a cell membrane or other kind of compartment?			According to the abstract, this manuscript covers a number of different areas relating to the RNA world hypothesis, including its compatibility with (likely) primitive Earth conditions; the likely characteristics of an RNA world; and, finally, the relationship between an early life-like RNA world system(s) and other life-like systems.			life6030029_perova	0
+If so, can RNA control the growth and division of the membrane?			Can we not have an RNA polymerase that uses other copies of itself as a template?			life6030029_perova	0
+If so, can RNA control the growth and division of the membrane?			These issues need to be considered separately.			life6030029_perova	0
+It is only mentioned in the introduction and conclusions.			Author Response Author Response File: Author Response.pdf			life6030029_perova	0
+"There are just a few issues that the authors should take into account for further improving their paper prior to acceptance: 1) For several references, the text ""Error!"			The article contains many original results of undeniable practical significance.			ma15051691_makarova	0
+Author Response Reviewer #2 (Comments to the Author): This manuscript relates to the effect of dysprosium substitution in Ce:YIG thin films and in particular to the temperature dependance of their Faraday rotation.			There are several questions about the content of the article.			ma15051691_makarova	0
+We answer the reviewer’s questions as follows.			demonstrate a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications.			ma15051691_makarova	0
+The saturation magnetization of rare-earth doped YIG affects the Faraday rotation angle. Sayetat et al. and Ostorero et al.			demonstrate a temperature stable magneto-optical material Dy2Ce1Fe5O12 for silicon integrated nonreciprocal photonic device applications.			ma15051691_makarova	0
+1) It is not indicated in the introduction whether any alternative to thin films of yttrium iron garnet is currently being considered for use as magneto-optical resonators. 2) How will the temperature dependence of the Faraday rotation hysteresis loops of Dy:CeYIG thin films change when moving to another wavelength range?			We answer the reviewer’s questions as follows.			ma15051691_makarova	0
+Please check with the editor if this can be fixed.			2) In equation (2) and the following line, there are inconcistencies in the use of capital letters for the Faraday rotation angles.			ma15051691_makarova	0
+The article contains many original results of undeniable practical significance.			It is shown that integrated MO isolators consist of Dy:CeIG thin films show better temperature stability compared to Ce:YIG based devices in the temperature range of 23 ℃ to 70 ℃.			ma15051691_makarova	0
+It is in general well written and the work deserves publication.			Ostorero, J.; Escorne, M.; Pecheron‐Guegan, A.; Soulette, F.; Le Gall, H. Dy3Fe5O12 garnet thin films grown from sputtering of metallic targets.			ma15051691_makarova	0
+Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?			It indeed the case, but I could not find any discussion on this issue.			ma15124258_makarova	0
+Every available suggestion has been answered carefully and corrected in the article.			-CL-20 crystal form is 622.7 nm-3 -CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.			ma15124258_makarova	0
+Are the onsets and finishing of the phase transformations consistent with the DSC measurements?			How are the cracks differentiated from the voids?			ma15124258_makarova	0
+How are the cracks differentiated from the voids?			“The electron density of ε-CL-20 crystal form is 622.7 nm-3 and that of γ-CL-20 is 584.8 nm-3, while the electron density of GPL107 is 571.7 nm-3” You need to specify the reference in which this data were obtained.			ma15124258_makarova	0
+Are the onsets and finishing of the phase transformations consistent with the DSC measurements?			I had several questions regarding this work: 1.			ma15124258_makarova	0
+Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.			ma15124258_makarova	0
+The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			As you can tell by these questions, my main issue is related to the specific form of the thermal damage.			ma15124258_makarova	0
+I recommend publishing the manuscript after minor the revision.			Could computed microtomography be used to confirm some of the experimental results?			ma15124258_makarova	0
+Some changes in propagation of pores caused by both the phase transitions and temperature were observed by an appropriate experimental technique.			How do these experiments differentiate between increasing the number of defects and the size of the defects?			ma15124258_makarova	0
+The conclusion must be supplemented with a forecast about the change in the performance of the CL-20 in terms of safety and suitability for use after temperature effects.			The DSC curve of CL-20 powders during thermal process Question 5.			ma15124258_makarova	0
+The authors used both WAXS and SAXS to examine CL-20 particles as they were heated from room temperature to around 200C.			If so, do the decomposition products form bubbles in your solvent?			ma15124258_makarova	0
+“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20.			To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”			ma15124258_makarova	0
+Response to Reviewer' Comments Dear Reviewer, We sincerely appreciate your comments and suggestions on the manuscript!			The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			ma15124258_makarova	0
+“During the thermal treatment, the nano-scale pores increase obviously, which will seriously increase the sensitivity of CL-20, and make a dangerous to the explosive charges with CL-20.			What do you mean by “are calculated” on line 199?			ma15124258_makarova	0
+I recommend publishing the manuscript after minor the revision.			The authors present an experimental study of nano-scale defects in CL-20 under heating.			ma15124258_makarova	0
+Do you have another proof for your conclusions.			Thus, I can recommend it for publication after a minor revision.			ma15124258_makarova	0
+What are the basic equations and assumptions for this calculation?			199-198 “The specific surface area and volume fraction of pores are calculated, and the results are shown in figure 7.” From the text of the manuscript it is not clear what method was used to obtain the calculated data on the specific surface area and volume fraction of pores.			ma15124258_makarova	0
+How do these experiments differentiate between increasing the number of defects and the size of the defects?			Could computed microtomography be used to confirm some of the experimental results?			ma15124258_perova	0
+), we described in detail how to calculate the specific surface area and volume fraction.			Can you explain why an increase in scattering intensity indicates increase in more small voids rather than an increase in existing void sizes?			ma15124258_perova	0
+Review of the manuscript Investigation on the Evolution of Nano-scale Defects of CL-20 Crystals under Thermal Treatment by Wide/Small-Angle X-ray Scattering, Haobin Zhang, Hongfan Wang … The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.			ma15124258_perova	0
+Therefore, I suggest include some comments on this and, probably, provide some discussion of how to avoid the increase of such defects, which increase sensitivity of energetic crystals, in particular CL-20.			As you can tell by these questions, my main issue is related to the specific form of the thermal damage.			ma15124258_perova	0
+), we described in detail how to calculate the specific surface area and volume fraction.			Overall, the work is done well and manuscript sounds.			ma15124258_perova	0
+Do you have another proof for your conclusions.			The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			ma15124258_perova	0
+Could the increase be due to larger defects rather than an increase in the number of defects?			To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”			ma15124258_perova	0
+Review of article entitled: “Investigation on the Evolution of Nano-scale Defects of CL-20 2 Crystals under Thermal Treatment by Wide/Small-Angle X-ray 3 Scattering” Overall, I liked the paper and believe it contributes to our knowledge of thermal evolution of CL-20.			Or, is it the volume of the defect that is increasing?			ma15124258_perova	0
+Could there be any other explanation rather than increase in number of internal nano-scale pores?			Are the onsets and finishing of the phase transformations consistent with the DSC measurements?			ma15124258_perova	0
+The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			In this paper, the same method was used to calculate the specific surface area and volume fraction, so we did not repeat how to calculator, just added the reference in the manuscript.			ma15124258_perova	0
+I recommend publishing the manuscript after minor the revision.			Are the onsets and finishing of the phase transformations consistent with the DSC measurements?			ma15124258_perova	0
+The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			How do these experiments differentiate between increasing the number of defects and the size of the defects?			ma15124258_perova	0
+The nano-scale defects were studied by using Wide Angle X-ray Scattering (WAXS) and Small Angle X-ray Scattering (SAXS), during the temperature range from 30 °C to 200 °C .			Every available suggestion has been answered carefully and corrected in the article.			ma15124258_perova	0
+The conclusion must be supplemented with a forecast about the change in the performance of the SL-20 in terms of safety and suitability for use after temperature effects.			Review of the manuscript Investigation on the Evolution of Nano-scale Defects of CL-20 Crystals under Thermal Treatment by Wide/Small-Angle X-ray Scattering, Haobin Zhang, Hongfan Wang … The manuscript is devoted to research on the phase transformations of CL-20 at various temperatures.			ma15124258_perova	0
+Overall, the work is done well and manuscript sounds.			Materials and instruments, you need to specify the particle size of the initial CL-20 powders.			ma15124258_perova	0
+To improve the application performance of CL-20, we should try to avoid the increase of such defects, such as storing in an constant low temperature to avoid the thermal expansion, and avoid any phase transition.”			), we described in detail how to calculate the specific surface area and volume fraction.			ma15124258_perova	0
+The authors used both WAXS and SAXS to examine CL-20 particles as they were heated from room temperature to around 200C.			Could computed microtomography be used to confirm some of the experimental results?			ma15124258_perova	0
+Is the omission of any items referring to experience/use of eCigarette/vaping.			The link between smoking and reduced life expectancy has been shown to be causal.			medicina58040502_makarova	0
+Response to Reviewer 1 I would like to thank Reviewer 1 for very detailed and useful comments which will definitely improve our manuscript.			The results section is too extensive - please avoid describing all the results in the text.			medicina58040502_makarova	0
+The results section is too extensive - please avoid describing all the results in the text.			Response to Reviewer 1 I would like to thank Reviewer 1 for very detailed and useful comments which will definitely improve our manuscript.			medicina58040502_makarova	0
+This is a well-designed and executed survey of university cigarette smoking in a country with above-average poverty.			The link between smoking and reduced life expectancy has been shown to be causal.			medicina58040502_makarova	0
+Medicina 2021, 57, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Lastname Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.			Lack of analysis of second-hand smoke exposure.			medicina58040502_makarova	0
+This is a well-designed and executed survey of university cigarette smoking in a country with above-average poverty.			In Table 5 please provide thr number of subjects (n=.....) Response 5.			medicina58040502_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			The link between smoking and reduced life expectancy has been shown to be causal.			medicina58040502_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			This is the opposite of health promotion one expects from all healthcare providers - especially physicians.			medicina58040502_makarova	0
+Author Response Please see the attachment Author Response File: Author Response.pdf			Results were consistent with previous studies of similar target populations.			medicina58040502_makarova	0
+"Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""			"Thus, exposure alone offers little information about the role of second-hand smoke and ""contagion."""			medicina58040502_makarova	0
+"The study aim should be more precise ""The aim of this study was...."" Response 1."			Please provide practical implications of this study Response 7.			medicina58040502_makarova	0
+Response to Reviewer 2 I would like to thank Reviewer 2 for very detailed and useful comments which will definitely improve our manuscript.			Tables 6 and 7 are unclear and should be more precise (Especially confidence intervals) Response 6.			medicina58040502_perova	0
+Healthcare providers who smoke provide a bad example to their patients - giving the impression that smoking is not dangerous.			Study sample - please provide more precise data on population (faculties at the university and its share in the total sample of recruited subjects) Response 2.			medicina58040502_perova	0
+Please attach an English version of the questionnaire as supplementary material or precisely describe questions that were used to assess the smoking status.			Medicina 2021, 57, x. https://doi.org/10.3390/xxxxx Academic Editor: Firstname Lastname Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.			medicina58040502_perova	0
+Please provide the most important results in the text, and the rest of them are present in Tables.			Results were consistent with previous studies of similar target populations.			medicina58040502_perova	0
+This is contrary to accepted public health policy internationally.			Specifically, whether exposure increases risk of smoking.			medicina58040502_perova	0
+It is likely that students who smoke actively seek roommates who smoke, and students who do not smoke seek roommates who do not smoke.			Author Response Please see the attachment Author Response File: Author Response.docx			medicina58040502_perova	0
+Thus it is unclear if students may be nicotine addicting by alternative means to tobacco smoking.			Finally, I found it disturbing that medical school faculty tolerate student smoking at all.			medicina58040502_perova	0
+In Table 5 please provide thr number of subjects (n=.....) Response 5.			Response to Reviewer 2 I would like to thank Reviewer 2 for very detailed and useful comments which will definitely improve our manuscript.			medicina58040502_perova	0
+This is a well-designed and executed survey of university cigarette smoking in a country with above-average poverty.			Author Response Please see the attachment Author Response File: Author Response.docx			medicina58040502_perova	0
+Is the omission of any items referring to experience/use of eCigarette/vaping.			This result calls into question the university's curriculum addressing the very many diseases associated with tobacco smoking - most of which reduce life expectancy.			medicina58040502_perova	0
+Please provide practical implications of this study Response 7.			This is the opposite of health promotion one expects from all healthcare providers - especially physicians.			medicina58040502_perova	0
+I suggest bringing this result up to the surface and making it more relevant even from the abstract.			Major comments: 1-My main issue is with the way the results are presented.			medsci4030012_makarova	0
+Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			Recurrent aberrations of 98 CNA regions were also identified in cases only.			medsci4030012_makarova	0
+It is not clear if there is one or two groups but just one percentage.			It is not clear which statistical method is used for the significant results.			medsci4030012_makarova	0
+2- The only real statistical inference they show is on the pathway analysis yet no mention is made on the abstract.			It is not clear which statistical method is used for the significant results in Table 2.			medsci4030012_makarova	0
+The authors need to give more details about how to use PCA in statistical methods.			The results are novel and provide insight into how multiple CNAs might contribute to cervical cancer development.			medsci4030012_makarova	0
+The authors mentioned the results by using PCA but PCA was not found in methods.			Broad comments This is the first genome-wide copy number analysis of CNAs identified for cervical cancer in the U.S. Latino population.			medsci4030012_makarova	0
+I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			Recurrent aberrations of 98 CNA regions were also identified in cases only.			medsci4030012_makarova	0
+Therefore, the authors pointed out that validation and confirmation of the results in a large sample size will be needed in the future.			Then again there must be estimates of deletion burden for different types of cancers from TCGA studies.			medsci4030012_makarova	0
+The results are novel and provide insight into how multiple CNAs might contribute to cervical cancer development.			The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			medsci4030012_makarova	0
+The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			The experimental design is adequate and the statistical methods are updated.			medsci4030012_makarova	0
+The authors have acknowledged the limitation of this study, for example, the small sample size.			The authors claim that they observed statistical differences between case and reference groups yet they do not preform a statistical test, nor give any p-value.			medsci4030012_makarova	0
+You can refer to it as methodological alternative, in the introduction, for the limited number of cases and lack of control group; issues that must be tackle in a larger study.			However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.			medsci4030012_makarova	0
+However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.			The ancestry of HapMap is not latino so therefore any test between cases and references will be confounded by ancestry.			medsci4030012_makarova	0
+They found that the burden of deletions in these patients is much higher than expected, as compared with CNV burden in HapMap populations.			A total of 32 CNA regions identified contained tumor suppressor genes and oncogenes.			medsci4030012_makarova	0
+Tobari and colleagues studied the CNV burden in patients with cervical cancer from Mexican American ancestry.			They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			medsci4030012_perova	0
+If you find good estimates then perhaps a statistical tests with those as your control group would be more believable.			[Response] great suggestions, thank the reviewer.			medsci4030012_perova	0
+However, I think that the manuscript needs some substantial revisions before it can be published.			Then again there must be estimates of deletion burden for different types of cancers from TCGA studies.			medsci4030012_perova	0
+They observed that their sample is enriched in CNV variants in pathways relevant to the phenotype like endometrial cancer and estrogen signaling pathway.			I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			medsci4030012_perova	0
+The authors need to define “pre-cancer” in materials.			Broad comments This is the first genome-wide copy number analysis of CNAs identified for cervical cancer in the U.S. Latino population.			medsci4030012_perova	0
+It is not clear which statistical method is used for the significant results.			The authors need to make clear about “pre-, cancer (91.3%) groups”.			medsci4030012_perova	0
+Tobari and colleagues studied the CNV burden in patients with cervical cancer from Mexican American ancestry.			Tobari and colleagues studied the CNV burden in patients with cervical cancer from Mexican American ancestry.			medsci4030012_perova	0
+The authors need to make clear about “pre-, cancer (91.3%) groups”.			They found statistically significant difference of CNA burden between case and reference groups in different sizes of CNAs that included deletions and amplifications, e.g., a statistically significant difference of >100 kb deletions were observed between the reference and pre-cancer and cancer groups.			medsci4030012_perova	0
+[Response] great suggestions, thank the reviewer.			The authors may need to delete the part in lines 238-242.			medsci4030012_perova	0
+The authors need to define “pre-cancer” in materials.			In the title it refers to Mexican American but in the rest of the manuscript it is treated as a latino population.			medsci4030012_perova	0
+They observed that their sample is enriched in CNV variants in pathways relevant to the phenotype like endometrial cancer and estrogen signaling pathway.			It is not clear if there is one or two groups but just one percentage.			medsci4030012_perova	0
+It is not clear if there is one or two groups but just one percentage.			However if the ancestry is only self reported as latino, then I would write latino in the title and mention the issue of genetic variability between latinos in the discussion.			medsci4030012_perova	0
+I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			Note that there are differences in admixture in the latino population depending of their country of origin.			medsci4030012_perova	0
+Minor comments: 1-I missed a definition of the sample's ancestry.			I think that these finding are indeed intriguing and deserve to be pursued in a larger study.			medsci4030012_perova	0
+In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			molecules26247651_makarova	0
+Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.			Check the font of the reference numbers in the text and correct			molecules26247651_makarova	0
+Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.			In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			molecules26247651_makarova	0
+Author Response Reviewer 1: Comments and Suggestions for Authors1.			There are some questions about the manuscript as follows: 1.			molecules26247651_makarova	0
+The cell values do not align please correct them.			Full characterization of all synthesized compounds is required in the experimental section.			molecules26247651_makarova	0
+In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			The explanation should be cleared.Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major: 1) The authors should give the reason for the nonlinear v(NH) vs delta correlation as it was (according to Dudek) linear for OH-O.			molecules26247651_makarova	0
+Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			Author Response Reviewer 1: Comments and Suggestions for Authors1.			molecules26247651_makarova	0
+Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			Author Response Reviewer 1: Comments and Suggestions for Authors1.			molecules26247651_makarova	0
+There are some questions about the manuscript as follows: 1.			"Minor: 183: should be SD = 34.3 cm-1186: """"are reasonable"""": Taking into account that (i) this fit is rather unphysical(ii) moving the points by say 50cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche."""			molecules26247651_makarova	0
+There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.2.			Author Response Reviewer 1: Comments and Suggestions for Authors1.			molecules26247651_makarova	0
+The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			molecules26247651_makarova	0
+Check the font of the reference numbers in the text and correct			The rationale for undertaking the research is a bit puzzling.			molecules26247651_makarova	0
+All the 1HNMR and 13CNMR must be attached as supplementary information.			There are some errors in spelling, syntax, punctuation, usage of capital letters, consistency in language style in the manuscript that must be improved.2.			molecules26247651_makarova	0
+In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.			molecules26247651_makarova	0
+There are some questions about the manuscript as follows: 1.			Scheme 1 must be redrawn using ACS document 1996 and all structures cleaned up.			molecules26247651_makarova	0
+It would be added value if these NH···O hydrogen bonds were also investigated in the solid state using XRD analysis.			I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			molecules26247651_perova	0
+The rationale for undertaking the research is a bit puzzling.			I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			molecules26247651_perova	0
+As indicated by the Reviewer, the present study is an extension of our previous OH···O binding studies.			3. All the 1HNMR and 13CNMR must be attached as supplementary information.			molecules26247651_perova	0
+8. Check the font of the reference numbers in the text and correct			6. Use 4 significant figures for the RNH values in table 2 7.			molecules26247651_perova	0
+6. Use 4 significant figures for the RNH values in table 2 7.			There are some questions about the manuscript as follows:1.			molecules26247651_perova	0
+Anyway, I think the article has some educational significance and could be published after few major and minor points are taken into account: Major:			Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			molecules26247651_perova	0
+In the experimental section of the 13C NMR it should be 125 MHz instead of 126 MHz.			Correct subscripts and superscripts in the experimental also check the spacings to be consistent.			molecules26247651_perova	0
+We have for some time wished to investigate the corresponding NH···O systems.			We have added a remark on this aspect to the Introduction section.			molecules26247651_perova	0
+"Minor: 183: should be SD = 34.3 cm-1186: """"are reasonable"""": Taking into account that (i) this fit is rather unphysical(ii) moving the points by say 50cm-1 one way or the other would also give rather good fit, this phrase is rather an empty cliche. Figure 5: Why is the y-axis reversed?"""			Rather, the author could check B3PW91 (it often performs better than B3LYP) or any newer suitable functional.			molecules26247651_perova	0
+In this article, the authors describe the spectral characteristics of NH in the hydrogen bridge NH...O.			Author Response Reviewer 1: Comments and Suggestions for Authors1.			molecules26247651_perova	0
+We have added a remark on this aspect to the Introduction section.			As indicated by the Reviewer, the present study is an extension of our previous OH···O binding studies.			molecules26247651_perova	0
+There are some questions about the manuscript as follows:			3. All the 1HNMR and 13CNMR must be attached as supplementary information.			molecules26247651_perova	0
+3. All the 1HNMR and 13CNMR must be attached as supplementary information.			There are some questions about the manuscript as follows:1.			molecules26247651_perova	0
+Author Response Reviewer 1The manuscript described ''The vibrational NH stretching transitions in secondary amines with intramolecular NH···O hydrogen bonds were investigated by experimental and theoretical methods, considering a large number of compounds and covering a wide range of stretching wavenumbers'' but the paper needs very significant improvement before acceptance for publication.			I am curious what prompted the Authors to deal with N-H...O bonding in place of the earlier O-H...O.			molecules26247651_perova	0
+[48] Accelrys Software Inc., Discovery Studio Modeling Environment, Release 4.0, San Diego: Accelrys Software Inc., 2013.			[26] Berrade, L.; Aisa, B.; Ramirez, M. J.; Galiano, S.; Guccione, S.; Moltzau, L. R.; Levy, F. O.; Nicoletti, F.; Battaglia, G.; Molinaro, G.; Aldana, I.; Monge, A.; Perez-Silanes, S. Novel Benzo[b]thiophene Derivatives as New Potential Antidepressants with Rapid Onset of Action.			molecules27051656_makarova	0
+Molecular Docking Study Molecular docking is a powerful tool to investigate and provide a proper understanding for ligand receptor interactions in order to facilitate the design of potential drugs [93-96].			It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].			molecules27051656_makarova	0
+The authors are suggested to use some decoy ligands and calculate the enrichment value in order to justify the docking protocol followed by ALA scan (Alanine scanning) based MD experiments.			13C NMR (100 MHz, DMSO-d6, ppm): 159.44 (C=O), 154.55 (Ar-C), 148.91 (Ar-C), 143.22 (-C=N-), 140.48 (Ar-C), 130.06 (Ar-C), 129.44 (Ar-C), 128.20 (Ar-C), 125.75 (Ar- C), 15.32 (-CH3).			molecules27051656_makarova	0
+Antimicrobial activity using agar disc diffusion method In vitro sensitivities of two gram-positive and two gram-negative bacteria including two fungal strains against the synthesized compounds were evaluated by agar disc diffusion method.			; Nahum Alves, C. Analysis of Kojic Acid Derivatives as Competitive Inhibitors of Tyrosinase: A Molecular Modeling Approach.			molecules27051656_makarova	0
+how would we know that −7.7 and −8.8 are good scores or not?			Moreover, ALA44A, LYS47A, ALA44B, LYS47B, PRO52A, ALA40B and ILE139B were actively involved in the non-covalent interaction (hydrophobic pi-alkyl).			molecules27051656_makarova	0
+A, 2002, 106, 10613-10621. https://doi.org/10.1021/jp0211786 [69] Li, Y.; Zhang, Y.; Niu, H.; Wang, C.; Qin, C.; Bai, X.; Wang, W. Schiff bases containing triphenylamine and pyrrole units: synthesis and electrochromic, acidochromic properties.			The authors did not report any Tyrosinase enzyme inhibition assay, they are requested to justify Tyrosinase inhibition as the probable mechanism of action for these sets of ligands by providing suitable literature references.			molecules27051656_makarova	0
+A., 2002, 106, 6871-6875. https://doi.org/10.1021/jp020715j [88] Scrocco, E.; Tomasi, J.			The manuscript is well-written and must be accepted in its present form for publication in Molecules.			molecules27051656_makarova	0
+An octanuclear [Co(II)2–Co(III)2]2 interlocked grid example of an inorganic [2]catenane†.			- Methods should be written in brief before presenting the results.			molecules27051656_makarova	0
+- This sentence “ ) — NHN=CH — CO — hydrazone moiety ( - containing hydrazide Polytopic ligands are important for new drug development” should be better connected to the next one to explain why they are important for new drug development?			It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic 3 adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme 97-103.			molecules27051656_makarova	0
+It is well-known that tyrosinase of Bacillus Megaterium bacteria is an attractive target for the development of antimicrobials or antibiotic adjuvants for the treatment of hyperpigmentation because of its similarity (33.5%) to the human enzyme [97-100].			PDB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium crystal structure.			molecules27051656_makarova	0
+Understanding the Regioselectivity on Diels−Alder Reactions.			The conclusion should briefly give an insight into the obtained results and also the limitations.			molecules27051656_makarova	0
+Why did the authors consider Tyrosinase from Bacillus megaterium N205A mutant pdb id 4j6u?			Two gram-positive Staphylococcus aureus (cars-2) and Bacillus Megaterium (BTCC-18), two gram-negative Escherichia coli (carsgn-2) and Salmonella Typhi (K-323130) bacteria, and two fungal strains Trichoderma harzianum (carsm-2) and Aspergillus niger (carsm-3) were used in this study.			molecules27051656_makarova	0
+5 Response: We appreciate the recommendation, and corrected this information as shown below: 3.2.			The 3D crystal structure of tyrosinase from Bacillus Megaterium (PDB ID: 4j6u; resolution: 2.5Å, Chain A, B) was obtained in pdb format from online RCSB protein data bank (PDB) database.			molecules27051656_makarova	0
+Each disc was loaded with 25 µL of sample solution in DMSO containing 300 µg of synthesized compounds.			1 Manuscript ID #: Molecules 1583723 Journal: Molecules A detailed point-by-point response to the reviewers' comments We would like to thank the reviewers for the comments and endorsements of our work.			molecules27051656_makarova	0
+The energy calculations were done by genetic algorithms.			[31] Ermiş, E. Synthesis, spectroscopic characterization and DFT calculations of novel Schiff base containing thiophene ring.			molecules27051656_makarova	0
+To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed.			New palladium(II) complexes of 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones.			molecules27051656_makarova	0
+Spect., 2014, 118, 672-682. http://dx.doi.org/10.1016/j.saa.2013.08.054 [71] Maryam, K.; Safar Ali, B.; Azar, G. Novel Schiff Bases of Pyrrole: Synthesis, Experimental and Theoretical Characterizations, Fluorescent Properties and Molecular Docking.			[42] Becke, A. D. Density‐functional thermochemistry.			molecules27051656_makarova	0
+To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain.			[20] Tikhomirov, A. S.; Litvinova, V. A.; Andreeva, D. V.; Tsvetkov, V. B.; Dezhenkova, L. G.; Volodina, Y. L.; Kaluzhny, D. N.; Treshalin, I. D.; Schols, D.; Ramonova, A.			molecules27051656_perova	0
+Struct., 2020, 1203, 127285. https://doi.org/10.1016/j.molstruc.2019.127285 [73] Cuenú, F.; Londoño-Salazar, J.; Torres, J. E.; Abonia, R.; D'Vries, R. F. Synthesis, structural characterization and theoretical studies of a new Schiff base 4-(((3-(tert-Butyl)-(1-phenyl) pyrazol-5-yl) imino) methyl) phenol.			Response: We appreciate the recommendation and rewrite the abstract as follows, 2 Polytopic organic ligands with hydrazone moiety are in the forefront of new drug research among many others due to their unique and versatile functionality and ease of strategic ligand design.			molecules27051656_perova	0
+To investigate and compare the antimicrobial activity of the synthesized compounds, docking analysis of L1 and L2 against tyrosinase from Bacillus Megaterium were performed.			Data Collect., 2020, 27, 100393. https://doi.org/10.1016/j.cdc.2020.100393 [60] Sundaram, M. S. S.; Karthick, S.; Sailaja, K.; Karkuzhali, R.; Gopu, G. Theoretical study on cyclophane amide molecular receptors and its complexation behavior with TCNQ.			molecules27051656_perova	0
+Chem., 2016, 40, 5245-5254. https://doi.org/10.1039/C6NJ00321D [70] Tanak, H.; Ağar A.			https://doi.org/10.1016/j.heliyon.2020.e04106 [59] Karpagakalyaani, G.; Daisy Magdaline, J.; Chithambarathanu, T.; Aruldhas, D.; Ronaldo Anuf, A. Spectroscopic (FT-IR, FT-Raman, NBO) Investigation and Molecular Docking study of a Herbicide compound Bifenox.			molecules27051656_perova	0
+The structure was verified, and an energy minimization was performed with the Swiss-Pdb Viewer software packages (version 4.1.0) [44], since the crystal structure contains a variety of issues related to improper bond order, side chains geometry, and missing hydrogen atoms.			DOI:10.33945/SAMI/AJCA.2020.4.14 [57] Balachandran, V.; Santhi, G.; Karpagam, V.; Lakshmi, A. Molecular structure, spectroscopic (FT-IR, FT-Raman), NBO and HOMO–LUMO analyses, computation of thermodynamic functions for various temperatures of 2, 6-dichloro-3-nitrobenzoic acid.			molecules27051656_perova	0
+Synthesis, structural, DFT studies, docking and antibacterial activity of a xanthene-based hydrazone ligand.			The docked conformation of the respective protein conformer with lowest binding free energy and root mean-square deviation value (RMSD) 0.0 Å was analyzed using PyMOL Molecular Graphics System (version 1.7.4) and Accelrys Discovery Studio 4.1 [49].			molecules27051656_perova	0
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation.			The binding site residues predicted by CASTp for tyrosinase were used for grid generation.			molecules27051656_perova	0
+PDB ID 4j6u is for tyrosinase from Bacillus megaterium, not the bacillus megatherium crystal structure.			Molecular negative and positive - Gram .’’ megaterium Bacillus ainst tyrosinase from molecular behavior of L1 and L2 ag Section 2.3.1.			molecules27051656_perova	0
+The binding site residues predicted by CASTp for tyrosinase were used for grid generation.			"explored ""Synthesis of Novel Tritopic Hydrazone Ligands: Spectroscopy, Biological activity, DFT, and Molecular docking Studies""."			molecules27051656_perova	0
+Even if it is an unstructured abstract, it should respond to the following point one after the other.			Author Response Manuscript ID #: Molecules 1583723 Journal: Molecules A detailed point-by-point response to the reviewers' comments We would like to thank the reviewers for the comments and endorsements of our work.			molecules27051656_perova	0
+To investigate the biological activities of the ligands, L1 and L2 were tested in vitro bioassays against some Gram-negative and Gram-positive bacteria and the fungus strain.			The data at least should be split into 2 more subheads, one for the ligand and receptor preparation and the second for the docking analysis.			molecules27051656_perova	0
+Enzyme Research, 2015, 2015, 1-12. http://dx.doi.org/10.1155/2015/262364 [99] Cardoso, R.; Valente, R.; Souza da Costa, C.H.			Attached below is our detailed response to the comments on our manuscript.			molecules27051656_perova	0
+The sample discs were put gently on pre-inoculated agar plates and aerobically incubated for 24 h at 37 °C for antibacterial and for 48 h at 26 °C for antifungal assay.			Synthetic Communications, 2020, 50(6), 831-839. https://doi.org/10.1080/00397911.2020.1720737 [94] Shweta; Khan, E.; Tandon, P.; Maurya, R.; Kumar, P. A theoretical study on molecular structure, chemical reactivity and molecular docking studies on dalbergin and methyldalbergin.			molecules27051656_perova	0
+All both compounds are biologically active but their activity was moderate which did not support their efforts.			Cancers, 2019, 11, 737. https://doi.org/10.3390/cancers11060737 [96] Yuan, Y.; Jin, W.; Nazir, Y.; Fercher, C.; Thomas Blaskovich, M.A.			molecules27051656_perova	0
+Dr. Md Abdul Majed Patwary Comilla University Author Response File: Author Response.pdf			In addition, Molecular docking was used to study the molecular revealed that outcomes The Bacillus megaterium.			molecules27051656_perova	0
+; da S. Gonçalves Vianez, J.L., Jr.; Santana da Costa, K.; de Molfetta, F.A.			Divide the data into those 2 subheads and add more about the software used to perform the docking and the parameters used (Grid box, extensiveness …) Response: We appreciate the recommendation, and splitted the data into 2 more subheads as, 3.2.1.			molecules27051656_perova	0
+This work presented the method for multiple metal ions detection using a carbon quantum dots (CQDs) based chemosensor array by functionalization with different amino acids.			So, I suggest to accept this paper to Molecule with only minor revision: Comments 1: In page 2, you write the heading “2.			molecules27123843_makarova	0
+Reviewer's comments and authors' replies: [Reviewer comments in black; Author response in blue.]			So, I suggest to accept this paper to Molecule with only minor revision: Comments 1: In page 2, you write the heading “2.			molecules27123843_makarova	0
+It is also impressive that all 11 metal ions were successfully identified by LDA with 100% classification rate.			Reviewer's comments and authors' replies: [Reviewer comments in black; Author response in blue.]			molecules27123843_makarova	0
+"Write ""On TEM images "" instead of ""Transmission electrone microscopy can be observed..."" Reply: Thanks for the valuable comment."			We truly appreciate your positive comments on our work, as well as for raising interesting points, which lead to the improvement of the manuscript.			molecules27123843_makarova	0
+Although the method of preparation of these sensors is not new, the implementation to chemosensor array is well presented.			This work presented the method for multiple metal ions detection using a carbon quantum dots (CQDs) based chemosensor array by functionalization with different amino acids.			molecules27123843_makarova	0
+Although the method of preparation of these sensors is not new, the implementation to chemosensor array is well presented.			We truly appreciate your positive comments on our work, as well as for raising interesting points, which lead to the improvement of the manuscript.			molecules27123843_perova	0
+-------------------------------------------------------------------- Response to Reviewer 1 This work presented the method for multiple metal ions detection using a carbon quantum dots (CQDs) based chemosensor array by functionalization with different amino acids.			Although the method of preparation of these sensors is not new, the implementation to chemosensor array is well presented.			molecules27123843_perova	0
+Reviewer's comments and authors' replies: [Reviewer comments in black; Author response in blue.]			Results”, but the content of that paragraph belong to the introduction part.			molecules27123843_perova	0
+The manuscript is an interesting research about developing a chemosensor array for metals ions detection simultaneously.			A detailed supplementary material is very helpful and useful for readers.			molecules27123843_perova	0
+The CQDs with and without metal ions were systematically characterized (TEM, IR, XRD, XPS, etc).			-------------------------------------------------------------------- Response to Reviewer 2 The manuscript is interesting research about developing a chemosensor array for metals ions detection simultaneously.			molecules27123843_perova	0
+From authors: Victor F. Tarasenko and Dmitry A. Sorokin Institute of High Current Electronics E-mail: VFT@loi.hcei.tsc.ru			One would expect a comparison between discharges that happen in gas mixtures of similar composition.			nano12040652_makarova	0
+Reply: The text of the article was finalized and the English was improved.			The topic is timely and the results provided are interesting.			nano12040652_makarova	0
+The article may be of interest, but for researchers in the field of plasma physics and gas discharges.			A point that should be clarified is the choice of the gases used for the discharges.			nano12040652_makarova	0
+This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc.			"For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."""			nano12040652_makarova	0
+Reply: This judgment was removed from the text of the manuscript.			Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow.			nano12040652_makarova	0
+However, most of the results provided consist in photographs of the discharges: while these are clear and descriptive, they provide a qualitative information only.			2) The article is more descriptive in nature, there is no explanation of physical laws, but which are misleading readers.			nano12040652_makarova	0
+Other gases, such as argon, were chosen to better demonstrate the discoloration of the discharge.			A point that should be clarified is the choice of the gases used for the discharges.			nano12040652_makarova	0
+In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material.			Reply: This judgment was removed from the text of the manuscript.			nano12040652_makarova	0
+The choice of the electrode material should be clearly explained in the present paper.			A spectral analysis is given for one discharge only (discharge in air with a copper electrode): more quantitative results, such as a spectral analysis of the other discharges also, would be helpful for the comparison between the discharges produced in the lab and the ones observed in the atmosphere.			nano12040652_makarova	0
+Reply: In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence.			Reply: In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence.			nano12040652_makarova	0
+Similar studies and comparisons with known experimental data could be carried out on these objects.			In spark or arc discharge, as well as in bright spots the electrodes are locally heated to a high temperature, which leads to the evaporation of the electrode material.			nano12040652_makarova	0
+We hope for a positive decision regarding the publication of this article in this journal.			So, when using electrodes made of aluminum, steel and copper, we observed the glow of red, blue and green colors, respectively.			nano12040652_perova	0
+The topic is timely and the results provided are interesting.			Reply: In our preliminary studies, as well as in the papers of other authors, it was found that during pulsed discharges of short duration, the electrode material determines the composition of the vapors that evaporate from the electrodes and diffuse into the gap, including due to shock waves and turbulence.			nano12040652_perova	0
+Lines 173-175 «Based on the brightness of the track glow, the plasma concentration at the particle surface increases with distance from the cathode.» Again, it is not clear how the plasma density on the particle surface can affect the brightness of the track glow.			Where does the evaporation of the metal take place?			nano12040652_perova	0
+Please describe better your experimental system, such as ignition energy.			In this regard, I would recommend that the authors revise the article (taking into account their colossal authority in the scientific community and the possibility of conducting more thorough experimental research) and send the article to a more specialized journal.			nano12040652_perova	0
+This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc.			We believe that an increase in the ra-diation intensity of the particle is due to its heating during deceleration on gas particles.			nano12040652_perova	0
+High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap.			One can agree with the influence of the particle size on the track size, but how can an increase in the particle charge affect the track glow?			nano12040652_perova	0
+"For instance: Lines 168, 169 ""An increase in the track brightness is apparently determined by an increase in the particle charge and size due to the evaporation of metal from the surface."""			High-temperature zones on the electrodes also supply micro- and nanoparticles into a discharge gap.			nano12040652_perova	0
+We hope that the results presented in the article will also be of interest to researchers who deal with micro and nanoparticles in traditional areas of nanomaterial physics [1-8].			Edits in the text are highlighted in either yellow or red font.			nano12040652_perova	0
+Reply to Reviewer 2 The article: «Influence of nanoparticles and metal vapors on the color of laboratory and atmospheric discharges» Authors: Victor Tarasenko, Nikita Vinogradov, Dmitry Beloplotov, Alexander Burachenko, Mikhail Lomaev, and Dmitry Sorokin Submission Date: 14 December 2021 Date of this review: 19 Jan 2022 07:18:39 Comments and Suggestions for Authors 1.			Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?			nano12040652_perova	0
+This is known from numerous studies of simpler and more understandable plasma objects such as glow discharge and arc.			Reply: The experimental system was described in more detail, and various modes of discharge ignition were analyzed.			nano12040652_perova	0
+Such inaccuracies are found throughout the text.			Is there any relationship between the specific choice of gases made by the authors and the composition of the atmosphere at the altitudes where the atmospheric discharges take place?			nano12040652_perova	0
+Not all abbreviations are listed for the table footnotes (FFM, D2O, DXA, LM, BV, ADP)			No technique is recommended, and the authors conclude that the methods are not interchangeable – not a novel finding.			nu14051073_makarova	0
+P2 L69: The aim stated here does not follow the logic of the preceding statements in the paragraph, i.e., what does comparing BC methods have to do with lack of healthcare or technology?			As stated above, it may be worth considering moving these Bland-Altman figures to the supplementary file.			nu14051073_makarova	0
+Desiree Lopez-Gonzalez Hospital Infantil de México Federico Gomez Dr. Márquez 162, Col. Doctores, Del.			"Relatedly, it would have been helpful to have more explanatory titles for the plots--eg ""Differential correlation between methods across levels of FM""."			nu14051073_makarova	0
+That is, what impact may underestimating FM, by relying on one particular method, have on child care?			One also begins to wonder about affecting type 1 errors due to multiplicity of comparisons (debatable but worth considering).			nu14051073_makarova	0
+If the focus (see the title) of the article is comparisons with the 4C model, then supplementary Table 1 seems to merit more direct attention.			Somehow this needs to be tied more firmly to the population and environment being studied.			nu14051073_makarova	0
+RESULTS Table 1: The n values in the column headers add up to 293 rather than 288.			That information may be helpful, perhaps also as supplementary data.			nu14051073_makarova	0
+The techniques are considered to be improvements over BMI for obesity assessment.			We have found your recommendations and commentaries of great value and have made appropriate changes.			nu14051073_makarova	0
+None of the methods are ideal – all are flawed (in comparison with a gold standard).			We have corrected the data, and now we only present data on the 288 subjects for both tables.			nu14051073_makarova	0
+We have added this in supplementary table as S table 1.			The second part of this analysis, titled: “Body composition assessment in Mexican children and adolescents.			nu14051073_makarova	0
+It seems that the authors are simply trying to point out that use of the 4C model is not common and that other methods are normally used.			P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).			nu14051073_makarova	0
+I didn't understand why the data differed between Table 1 and Supplementary Table 1.			The aim (and intent) of the study seems too simplistic as stated here – merely comparing methods.			nu14051073_makarova	0
+The data used was total body with head, as recommended for the ISCD when using DXA for body composition instead of for bone densitometry.			of the paper is all about comparing techniques against the 4C model.			nu14051073_makarova	0
+Part 2: Cross-validation of three bio-electrical impedance methods against dual X-ray absorptiometry for whole-body and regional body composition”, has already been accepted by Nutrients #1604578.			We have corrected the data, and now we only present data on the 288 subjects for both tables.			nu14051073_makarova	0
+of the paper is all about comparing techniques against the 4C model.			Table 1: The n values in the column headers add up to 293 rather than 288.			nu14051073_makarova	0
+Reviewer two comments: I am not familiar with the Bland-Altman method, so was unclear when I read the methods section whether the text in lines 204-207 was explaining the Bland-Altman method or describing a separate procedure.			We have found your recommendations and commentaries of great value and have made appropriate changes.			nu14051073_makarova	0
+This ranking is itself very valuable in routine clinical care, particularly for longitudinal assessment.			P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).			nu14051073_makarova	0
+This ranking is itself very valuable in routine clinical care, particularly for longitudinal assessment.			Added, the analysis was for the total sample and by subgroups by age and sex, line 287 P12 L283: Is Table 4 necessary?			nu14051073_makarova	0
+P7 L226: It would be helpful if it was made clear that this refers to comparisons of means of all subjects (not broken down by age, sex).			Desiree Lopez-Gonzalez Hospital Infantil de México Federico Gomez Dr. Márquez 162, Col. Doctores, Del.			nu14051073_makarova	0
+Also, the difference(s) between FFM and LM definitions are nuanced and complex and should either be defined, or alternate text should be used here.			I hope that you find this work suitable for your Journal and I look forward to hearing from you.			nu14051073_makarova	0
+Response to reviewers' comments: Reviewer one: TITLE The “Part 1” in the title, while perhaps intriguing, calls for some explanation of future directions.			Hospital Infantil de México Federico Gómez Instituto Nacional de Salud UNIDAD DE EPIDEMIOLOGÍA CLÍNICA Hospital Infantil de México Federico Gómez Instituto Nacional de Salud UNIDAD DE EPIDEMIOLOGÍA CLÍNICA Mexico City, February 23, 2022 To the Editorial Board and Expert Reviewers: We, the authors, want to express our gratitude to the Editorial Board and the experts that reviewed our manuscript for their contributions.			nu14051073_makarova	0
+Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD.			Because the KQ-10 is a composite tests there is the likelihood to be some interaction with the sub-domains.			nu14122489_perova	0
+The proposal about “Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents” is attractive.			Use all five of the KO-10 dimensions as your outcome measure not the Mean average global KQ-10 score.			nu14122489_perova	0
+Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet.			In table 5 only one p value is reported, but what it is measuring is unclear, as there are a number of interactions occurring.			nu14122489_perova	0
+-Conclusion and discussion: They are well elaborated.			2020 Jul;23(10):1754-1765. doi: See their Table 5.			nu14122489_perova	0
+Journal of nutritional science and vitaminology, 61(Supplement), S66-S68.			The conclusion is sound given the findings but the lack of reference to income is an issue as educational status of parents is often a “de-facto” measure for income.			nu14122489_perova	0
+It is more that education it is also income re line 106-107 “It is proposed that parents with more educational experience are more likely to make healthier food choices for their family” .			Remember the KQ-10 is made up of five subdomains (psychological well-being, autonomy and parent relation, social support and peers, and school environment) The fact is diet is no expected to change 3 of these KQ-3 subdomains and only one or two KQ-10 dimensions.			nu14122489_perova	0
+It maybe, it is the cost of living associated with fresh fruit and vegetables that is the real issue.			The European Journal of Health Economics, 15(9), 967-977.			nu14122489_perova	0
+Similarly, a correlation matrix is typically reported as it the foundation of regression analyses and so it needs to be reported to understanding the interaction between three main tests variables being investigated in this study.			The important issue is in the method section the reader needs understand the dimensions and sub-dimension that make up within each survey.			nu14122489_perova	0
+Reviewer 1: The proposal about “Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents” is attractive.			The lack of recommendations associated with the Swedish free lunch program to assist the diets of students in low SES families to have more fruit and vegetables			nu14122489_perova	0
+Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			Remember the KQ-10 is made up of five subdomains (psychological well-being, autonomy and parent relation, social support and peers, and school environment) The fact is diet is no expected to change 3 of these KQ-3 subdomains and only one or two KQ-10 dimensions.			nu14122489_perova	0
+Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents Overall comments: supportive of the paper but have concerns about it for the following 5 reasons.			Diet is not expected to have any influence of parent relations, social relations or peers but your study may find an influence on psychological well-being and even school environment.			nu14122489_perova	0
+The following paper did a better job of reporting it Moraeus L, Lindroos AK, Warensjö Lemming E, Mattisson I.			If income is not important than the paper should be the on home education level and home diet not home SES and home diet.			nu14122489_perova	0
+Diet diversity score and healthy eating index in relation to diet quality and socio-demographic factors: results from a cross-sectional national dietary survey of Swedish adolescents.			Given you are using education of parents as the status measure of SES the term status is important in this paper and should not be dropped.			nu14122489_perova	0
+Focusing only on the global KQ-10 scores is hiding the subdomain differences to diet.			Bouwmans, C., van der Kolk, A., Oppe, M., Schawo, S., Stolk, E., van Agthoven, M., ... & van Roijen, L. (2014).			nu14122489_perova	0
+I recommend you to alternate them (text, Figures and Tables in Results).			To improve, you need the next questions: -Abstract: it is not conventional.			nu14122489_perova	0
+To be prudent, try to update some references if you find.			Given you are using education of parents as the status measure of SES the term status is important in this paper and should not be dropped.			nu14122489_perova	0
+Validity and responsiveness of the EQ-5D and the KIDSCREEN-10 in children with ADHD.			2020 Jul;23(10):1754-1765. doi: See their Table 5.			nu14122489_perova	0
+Unless these are included the reader can not fully understand the study.			-Conclusion and discussion: They are well elaborated.			nu14122489_perova	0
+For while gender and in this study education are categorical (group) the data being evaluated is continuous and interval data and so an ANOVA or MANOVA by group is the method of analysis of variance.			Given you are using education of parents as the status measure of SES the term status is important in this paper and should not be dropped.			nu14122489_perova	0
+A public education program could also be encouraged about health eating.			Bouwmans, C., van der Kolk, A., Oppe, M., Schawo, S., Stolk, E., van Agthoven, M., ... & van Roijen, L. (2014).			nu14122489_perova	0
+The basic information is not there in the paper: “KIDSCREEN-10 (KS-10) is derived from the KIDSCREEN-27, and provides a single index of global QoL using ten items related to physical well-being, psychological well-being, autonomy and parent relation, social support and peers, and school environment” See how these researchers have described the KIDSCREEN-10 .			Reviewer 1: The proposal about “Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents” is attractive.			nu14122489_perova	0
+Reviewer 2: Open Review Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents Overall comments: supportive of the paper but have concerns about it for the following 5 reasons.			If income is not important than the paper should be the on home education level and home diet not home SES and home diet.			nu14122489_perova	0
+The Swedish Healthy Eating Index for Adolescents 2015 (SHEIA15) is not well explained in this paper.			Effects of socioeconomic status on nutrition in Asia and future nutrition policy studies.			nu14122489_perova	0
+Bouwmans, C., van der Kolk, A., Oppe, M., Schawo, S., Stolk, E., van Agthoven, M., ... & van Roijen, L. (2014).			If the focus is on gender typically both a boy and a girl regression model is reported.			nu14122489_perova	0
+The first sentence assumes that poor diet is always associated with adolescents, this is incorrect, Better to say: Adolescents often do not consume a high-quality diet, concurrently their self- 13 reported mental health problems are increasing			It maybe, it is the cost of living associated with fresh fruit and vegetables that is the real issue.			nu14122489_perova	0
+Line 90 need to put in the full name Health Related Quality of Life (HRQoL).			To improve, you need the next questions: -Abstract: it is not conventional.			nu14122489_perova	0
+To assist the reader’s comprehension, explain the instruments more as you develop the paper.			Similarly, a correlation matrix is typically reported as it the foundation of regression analyses and so it needs to be reported to understanding the interaction between three main tests variables being investigated in this study.			nu14122489_perova	0
+It is more that education it is also income re line 106-107 “It is proposed that parents with more educational experience are more likely to make healthier food choices for their family” .			The following paper did a better job of reporting it Moraeus L, Lindroos AK, Warensjö Lemming E, Mattisson I.			nu14122489_perova	0
+The proposal about “Investigation of the Associations between Diet quality and Health-Related Quality of Life in a Sample of Swedish Adolescents” is attractive.			I am a not European reviewer and so found the RADDS a rather restricted list without some meat.			nu14122489_perova	0
+The topic is interesting, and the manuscript is clear.			Comment 3: Please revise Figure 1 because it's very difficult to read.			obesities2010010_makarova	0
+This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			Comment 3: Please revise Figure 1 because it's very difficult to read.			obesities2010010_makarova	0
+The topic is interesting, and the manuscript is clear.			The topic is interesting, and the manuscript is clear.			obesities2010010_makarova	0
+This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI.			The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach.			obesities2010010_makarova	0
+The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).			obesities2010010_makarova	0
+The topic is interesting, and the manuscript is clear.			Comment 3: Please revise Figure 1 because it's very difficult to read.			obesities2010010_makarova	0
+This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			I have some comments for the authors about the structure of the manuscript that should take into consideration.			obesities2010010_makarova	0
+The manuscript by Grant el al., is an interesting study in which they exhaustively utilized a very large database generated for almost 3 decades to analyze the representation of obesity and public health policy and its association with gender, healthiness, social status, and negative stereotypes by using machine learning and computational language analysis approach.			This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			obesities2010010_makarova	0
+Author Response Reviewer 1: General comments:			You report that an automatic approach was used to check all the papers.			obesities2010010_makarova	0
+Author Response Reviewer 2: General comments: The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			Author Response Reviewer 1: General comments:			obesities2010010_perova	0
+A recent paper has pointed out that postbariatric patients, using a novel approach based on words, presented a weight bias regards their body, but it was not present for other bodies (see http://dx.doi.org/10.1007/s11695-020-05166-z).			The topic is interesting, and the manuscript is clear.			obesities2010010_perova	0
+I think this aspect is interesting and linked to your results, showing a connection with clinical data.			The manuscript reports a study about weight-biased language in the Australian media across the last decades.			obesities2010010_perova	0
+The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			Is there any possible limit with this approach in the selection of the papers?			obesities2010010_perova	0
+I do not have particular concerns about the paper.			This is interesting topic that, although based on restricted area exploration, may have much broader significance in analyses that reveal worldwide social trends, as a whole.			obesities2010010_perova	0
+The methods applied are innovative and allow to underline new interesting evidence.			Author Response Reviewer 2: General comments: The authors present a manuscript which evaluates weight-biased language across 30 years of Australian news reporting on obesity: Associations with public health policy.			obesities2010010_perova	0
+I do not have particular concerns about the paper.			I think this aspect is interesting and linked to your results, showing a connection with clinical data.			obesities2010010_perova	0
+I would like the authors to emphasize the impact of the analyzed trends by means of mental disorders, at least to those with significant clinical and medical importance.			This study highlights a very relevant topic of obesity and weight stigma in the society, particularly when the research have shown that “metabolically obese normal weight” people still exhibit significant risk of developing cardiovascular and other metabolic related disorders, regardless of having normal weight and BMI.			obesities2010010_perova	0
+Therefore, I suggest exploring, using the same methodology, the data regarding the associations of weight-biased language with eating disorders (the incidence of anorexia nervosa and bulimia) and mood disorders (the incidence of depression and anxiety, etc.).			Author Response Reviewer 3: General comments: The manuscript reports a study about weight-biased language in the Australian media across the last decades.			obesities2010010_perova	0
+The work is not unprecedented in the concept, but provides interesting breeding material for specific indian farmers, which is an important achievement and is correctly taken up by authorities and international research centers.			Then the table can be more informative and ease to read.			plants11050622_makarova	0
+The paper itself is well written, although 1) somewhat results are partially descriptive and partially inferential.			This work is of outstanding quality, and I normally present more critical points in my reviews.			plants11050622_makarova	0
+Table and main text are independent, so the authors have to describe the table more carefully.			Legends should describe what is to be observed in the figure, possibly pointing to parts of special interest.			plants11050622_makarova	0
+On Supplementary table S2, does significant mean p-value?			Indicate DFF = days to fifty percent flowering.			plants11050622_makarova	0
+Correct it It was a pleasure to read this manuscript.			The last paragraph of results is about background selection.			plants11050622_makarova	0
+Author Response Point-by-point response to reviewer's comments			How did you calculate those values on those comparison types?			plants11050622_makarova	0
+The data on bgs are not sufficiently presented in the text nor discussed.			Author Response Point-by-point response to reviewer's comments			plants11050622_makarova	0
+The corrections have been incorporated as per the suggestions made by the reviewer.			As such, it is an overview paper that raises many questions.			plants11050622_makarova	0
+is probably not clear and ILs no should maybe considered instead.			Correct it It was a pleasure to read this manuscript.			plants11050622_makarova	0
+It is also acknowledged that this paper is probably the first of many papers to emerge from the study.			How did you calculate those values on those comparison types?			plants11050622_makarova	0
+This paper described the multi-parents introgression assisted by molecular markers.			This paper described the multi-parents introgression assisted by molecular markers.			plants11050622_makarova	0
+Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups.			The last paragraph of results is about background selection.			plants11050622_makarova	0
+The authors should consider reshaping the tables, eliminating unnecessary columns (e.g.			The work is not unprecedented in the concept, but provides interesting breeding material for specific indian farmers, which is an important achievement and is correctly taken up by authorities and international research centers.			plants11050622_makarova	0
+L127: Please add the explanation of “boro season”.			L296-L302: the marker descriptions on main text cannot match the supplementary table S8.			plants11050622_makarova	0
+With minor grammatical revisions, the manuscript can be accepted as is.			The work is not unprecedented in the concept, but provides interesting breeding material for specific indian farmers, which is an important achievement and is correctly taken up by authorities and international research centers.			plants11050622_makarova	0
+Correct it It was a pleasure to read this manuscript.			Line 262: “=261 %” , and Line 312 “linkes” a typo?			plants11050622_makarova	0
+The phenotypic data are important, but currently they are presented only as additional material.			The last paragraph of results is about background selection.			plants11050622_makarova	0
+Then the table can be more informative and ease to read.			Author Response Point-by-point response to reviewer's comments			plants11050622_makarova	0
+The authors should consider reshaping the tables, eliminating unnecessary columns (e.g.			However, the authors have conducted a thorough literature review, undertaken a rigorous piece of data collection, and have generalized information accurately.			plants11050622_makarova	0
+How did you calculate those values on those comparison types?			Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups.			plants11050622_makarova	0
+This work is of outstanding quality, and I normally present more critical points in my reviews.			I only recommend some minor revisions before acceptance.			plants11050622_makarova	0
+Please add some gel pictures of the foreground selection markers you used in order to visualize the genotyping results and showed the polymorphism of these markers on gel.			This paper described the multi-parents introgression assisted by molecular markers.			plants11050622_makarova	0
+Also, the legends are not clearly describing all the terms and should be revised.			is probably not clear and ILs no should maybe considered instead.			plants11050622_makarova	0
+Please re-organize the Supplementary table S5, I suggest use each ILs only appear one time and add one more column to show their PC groups.			Author Response Point-by-point response to the reviewer's comments are given below The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			plants11050622_makarova	0
+L48-63: should add some citations on the first part of introduction.			The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			plants11050622_makarova	0
+L562: the section of statistical analysis should add more details.			As such, it is an overview paper that raises many questions.			plants11050622_makarova	0
+On Supplementary table S2, does significant mean p-value?			How did you calculate those values on those comparison types?			plants11050622_makarova	0
+of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion.			The legend of supplementary table should be improved.			plants11050622_makarova	0
+The work is not unprecedented in the concept, but provides interesting breeding material for specific indian farmers, which is an important achievement and is correctly taken up by authorities and international research centers.			I only recommend some minor revisions before acceptance.			plants11050622_makarova	0
+of genes / QTLs, which is already represented in column 1) and add the most striking phenotypic data, when possible and relevant for discussion.			The ms is the result of an intensive and years-long work of breeding, that eventually pyramidized several resistance genes and QTLs for abiotic traits into an indian elite rice variety.			plants11050622_makarova	0
+The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			What characterisation was performed to confirm the identity of cells as ESCs?			reprodmed3020015_makarova	0
+SEM is a function of how many measurements were made, and does not reflect sample variability.			It is important to determine whether these mechanisms are both operating.			reprodmed3020015_makarova	0
+Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1.			The western blot (also Fig.3) result must show the entire molecular weight range (e.g.			reprodmed3020015_makarova	0
+If siRNA-resistant PGRMC1 levels are able to inhibit decidualization it would provide convincing evidence in favour of the hypothesis that lowered PGRMC1 is required for decidualization, and is perhaps even a trigger.			Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) Lines 271-76.			reprodmed3020015_makarova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			If further mechanism exploration is carried out, the research will be more complete and persuasive.			reprodmed3020015_makarova	0
+Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool.			The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough.			reprodmed3020015_makarova	0
+reflects variability between replicate measurements.			The conclusion has been verified by other 2-3 articles, and the design of this study is not rigorous enough.			reprodmed3020015_makarova	0
+4) Lines 263-267: “Because the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence.			This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			reprodmed3020015_makarova	0
+In general, there is very little detail on these primary cultured lines.			At the very least this mechanism merits detailed discussion in that section.			reprodmed3020015_makarova	0
+The authors have addressed most of my concerns.			Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1.			reprodmed3020015_makarova	0
+Appropriate human ethics approvals were obtained.			The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			reprodmed3020015_makarova	0
+It is important to determine whether these mechanisms are both operating.			What characterisation was performed to confirm the identity of cells as ESCs?			reprodmed3020015_makarova	0
+Line 207: “The miRNA target prediction database, miRDB (http://www.mirdb.org), was screened to identify miRNAs with potential to interact with PGRMC1.” miR-98 is already known to regulate PGRMC1.			We are told in the methods that primary cell cultures were obtained from 3 patients (see lines 88-89).			reprodmed3020015_makarova	0
+Author Response Responses to Reviewer 2 We appreciate your review of our manuscript and constructive comments and suggestions.			The manuscript explored the mechanisms of PGRMC1 regulation during the decidualization of human ESCs.			reprodmed3020015_makarova	0
+And please provide basic information of the patient.			The authors must refer to the fact that AG-205 is not specific for PGRMC1, contrary to many citations in the literature (such as would be the case here, if uncorrected).			reprodmed3020015_makarova	0
+Such changes may occur at the translational or post-translational level.			However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4.			reprodmed3020015_makarova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			reprodmed3020015_makarova	0
+It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1.			Line 261. mechanisms HAVE not (plural mechanisms) Lines 261-262.			reprodmed3020015_makarova	0
+Cite PMID: 34944026, PMID: 34680104, PMID: 32924377.			The authors mention technical difficulties in establishing ESCs that express miRNA-resistant PGRMC1.			reprodmed3020015_makarova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualization.			The paragraph describes a result showing that the results are independent of P4.			reprodmed3020015_makarova	0
+Appropriate human ethics approvals were obtained.			Aspiration to clarify this in future papers does not merit publication of the present paper.			reprodmed3020015_makarova	0
+Downregulation of PGRMC1 During ESC in vitro Decidualization.” This seems to be the next section heading, in which case it should follow a line break, and say: “3.2.			It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1.			reprodmed3020015_makarova	0
+SEM is a function of how many measurements were made, and does not reflect sample variability.			The new text does not conceptually link PTMs with PGRMC1 protein stability, which is the only reason they should be mentioned.			reprodmed3020015_makarova	0
+We are told in the methods that primary cell cultures were obtained from 3 patients (see lines 88-89).			In response the authors have made minimal changes to the manuscript, effectively restricted to rewording the title, and several minor text changes.			reprodmed3020015_makarova	0
+Assuming binomial distribution, a two-way ANOVA is required for these experiments.			PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988).			reprodmed3020015_makarova	0
+It is suggested to add cell lines or normal endometrial cells for verification.			Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test.			reprodmed3020015_makarova	0
+4) Lines 263-267: “Because the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence.			Critically, these figures compare the effects of two variables each, and therefor one way ANOVA is not the appropriate test.			reprodmed3020015_makarova	0
+However, as described above AG-205 is not a specific inhibitor of PGRMC1.			If higher PGRMC1 levels are indeed critical for the suppression of decidualization, as is reasoned by the authors, then it should be possible to maintain PGRMC1 levels artificially to inhibit decidualization, as discussed below.			reprodmed3020015_makarova	0
+Statistical sample sizes were unfavourably small.			It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.			reprodmed3020015_makarova	0
+Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation.			Therefore, a resubmission after major revision is recommended.			reprodmed3020015_makarova	0
+We are told in the methods that primary cell cultures were obtained from 3 patients (see lines 88-89).			The western blot (also Fig.3) result must show the entire molecular weight range (e.g.			reprodmed3020015_makarova	0
+PGRMC1 protein instability could be the dominant effect involved, with miRNA transcript regulation playing only a minor part.			Figure 2 seems to depict just one cell culture (which is not even named), with 4 replicates of each measurement.			reprodmed3020015_makarova	0
+The research has a certain degree of scientific significance.			And please provide basic information of the patient.			reprodmed3020015_makarova	0
+The fact that PGRMC1 levels fall does not mean that low PGRMC1 levels are required.			That new experimental result is what elevated the required changes to a major revision.			reprodmed3020015_makarova	0
+Give the catalogue number of the secondary antibody.			If higher PGRMC1 levels are indeed critical for the suppression of decidualization, as is reasoned by the authors, then it should be possible to maintain PGRMC1 levels artificially to inhibit decidualization, as discussed below.			reprodmed3020015_makarova	0
+Were primary cultures used directly, or were they stored cryogenically?			What percentage acrylamide were the SDS PAGE gels.			reprodmed3020015_makarova	0
+It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).			Which ImageJ plug-in was used for quantification?			reprodmed3020015_makarova	0
+Author Response We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript.			This could be accomplished by making an expression plasmid encoding a codon-altered PGRMC1 gene, and cotransfecting this in a controlled matrix design with siRNA.			reprodmed3020015_makarova	0
+It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).			Author Response We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript.			reprodmed3020015_makarova	0
+This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues.			The manuscript explored the mechanisms of PGRMC1 regulation during the decidualization of human ESCs.			reprodmed3020015_makarova	0
+“Data of cell culture are expressed as the mean ± SD from three independent experiments with at least repeated two times.” Change to: Data of cell culture are expressed as the mean ± SD from three independent experiments.			Sensitivity of a phenomenon to AG-205 is consistent with possible PGRMC1 involvement, but does not demonstrate PGRMC1 involvement.			reprodmed3020015_makarova	0
+The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.			It would be possible to introduce an expression cassette via a lentiviral vector at MOI 1.			reprodmed3020015_perova	0
+And please provide basic information of the patient.			Assuming binomial distribution, a two-way ANOVA is required for these experiments.			reprodmed3020015_perova	0
+The discussion must qualify the results of Salsano et al.			PGRMC1 (Hpr6.6) increased the rate of cell death (in a non-apoptotic mechanism) in MCF-7 cancer cells in response to H2O2 (PMID: 14523988).			reprodmed3020015_perova	0
+Unfortunately, the quality of experimental design and results processing are somewhat problematic.			Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			reprodmed3020015_perova	0
+Author Response Responses to Reviewer 1 We appreciate your review of our manuscript and constructive comments and suggestions.			Comparison of the miR-98/PGRMC1 abundance in proliferative and secretory phases of endometrial biopsy may contribute to the diagnostic prediction of uterine receptivity for embryo implantation.			reprodmed3020015_perova	0
+Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			Our responses to your comments are as follows.			reprodmed3020015_perova	0
+“In the present study, we showed that microRNA-mediated PGRMC1 regulation during the process of decidualization.” This is confusingly worded.			This new text should be discussing the possibility of the latter also contributing to the observed effects, as well as the effects of mRNA level that the paper pursues.			reprodmed3020015_perova	0
+Were primary cultures used directly, or were they stored cryogenically?			This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualisation.			reprodmed3020015_perova	0
+Which ImageJ plug-in was used for quantification?			It is much preferable to refer to protein abundance, and changes in abundance.			reprodmed3020015_perova	0
+However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4.			Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			reprodmed3020015_perova	0
+I am uncertain because the methods do not explain how the experiment was performed.			At the very least this mechanism merits detailed discussion in that section.			reprodmed3020015_perova	0
+Such vectors typically include fluorescent protein reporters, enabling discrimination between infected and non-infected cells in one plate, or population separation by FACS.			The simplest direct comparison would be db-cAMP/P4 vs. db-cAMP to permit the argument the authors are presenting.			reprodmed3020015_perova	0
+Since that is by no means certain, the study should not be published in its present form.			The introduction succinctly provides background relevant for the following research description.			reprodmed3020015_perova	0
+Therefore, a resubmission after major revision is recommended.			Appropriate human ethics approvals were obtained.			reprodmed3020015_perova	0
+The authors go on to show that expogenous expression of miR-98 lowers PGRMC1 abundance levels (a more apt description than the fuzzy description of “PGRMC1 downregulation”, which could be by a number of mechanisms as discussed below).			Such changes may occur at the translational or post-translational level.			reprodmed3020015_perova	0
+The experiemtnal design for this conclusion is problematic, as described below.			Translational level changes can be due to changed levels of transcription, to altered mRNA stability (such as that proposed here for miR-98), or to altered efficiency of translation of a steady state mRNA pool.			reprodmed3020015_perova	0
+The latter could well contribute to alterations in PGRMC1 abundance cited by this study.			The latter could well contribute to alterations in PGRMC1 abundance cited by this study.			reprodmed3020015_perova	0
+In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			It is much preferable to refer to protein abundance, and changes in abundance.			reprodmed3020015_perova	0
+Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy.			Which ImageJ plug-in was used for quantification?			reprodmed3020015_perova	0
+PGRMC1 levels are also controlled by proteolysis, mediated by the ubiquitin proteasome system as described above.			Before such a claim is published in the scientific literature, it must be supported by convincing evidence.			reprodmed3020015_perova	0
+It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.			Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy.			reprodmed3020015_perova	0
+PGRMC1-independe (add t) Lines 263-267: “Because, the effect of AG-205 and PGRMC1 knock-down on decidualization was common in the present study, these findings further support for a role for PGRMC1 downregulation in promoting ESC decidualization during the secretory phase.” The reviewer does not understand the sentence.			Immunocytochemistry (ICC) of vimentin and cytokeratin in isolated human ESCs.			reprodmed3020015_perova	0
+Downregulation of PGRMC1 During ESC in vitro Decidualization.” All subsequent section header numbers in results should be modified to accommodate the new section.			An inducible vector may be advisable, such as tetracycline, IPTG, or other available systems.			reprodmed3020015_perova	0
+What characterisation was performed to confirm the identity of cells as ESCs?			Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) Lines 271-76.			reprodmed3020015_perova	0
+Suggest change to; “Further study is required to determine the relationship between miR-98-mediatd PGRMC1 regulation and pregnancy.			However, in the first instance the manuscript cannot be published as it stands, and should therefore be rejected now with the stated option of subsequent resubmission only if new evidence is obtained.			reprodmed3020015_perova	0
+If PGRMC1 down-regulation is critical, it should be possible to perturb decidualization by expression of an exogenous PGRMC1 protein.			Such changes may occur at the translational or post-translational level.			reprodmed3020015_perova	0
+It is also suggested throughout to use “AG-205” (the correct name for the reagent: https://www.sigmaaldrich.com cat#: a1487) rather than AG205 (often incorrectly used in the literature).			Suggest change to “In the present study, we showed that microRNA-mediated the regulation of PGRMC1 abundance during the process of decidualization.” (if this is what is meant) Lines 271-76.			reprodmed3020015_perova	0
+However, the effects of P4 in the absence of db-cAMP would also require control in cells without db-cAMP, +/- P4.			The authors go on to show that expogenous expression of miR-98 lowers PGRMC1 abundance levels (a more apt description than the fuzzy description of “PGRMC1 downregulation”, which could be by a number of mechanisms as discussed below).			reprodmed3020015_perova	0
+An inducible vector may be advisable, such as tetracycline, IPTG, or other available systems.			Our responses to your comments are as follows.			reprodmed3020015_perova	0
+Individual mutation of these residues could stabilise PGRMC1 levels to reduce decidualization if miR-98 is only partially responsible for reducing PGRMC1 levels.			It is important that the involvement of the ubiquitin pathway contribution be either quantified or discarded in well controlled experiments to justify the title of the paper.			reprodmed3020015_perova	0
+Our responses to your comments are as follows.			Appropriate human ethics approvals were obtained.			reprodmed3020015_perova	0
+The methods could describe that all subjects were ethnically Japanese/Asian (or otherwise).			Suggest observations that may be able to discriminate between different possibilities.			reprodmed3020015_perova	0
+In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			demonstrate that PGRMC1 abundance levels are higher in proliferative phase versus secretory phase endometrial patient biopsies (fig.1).			reprodmed3020015_perova	0
+Our responses to your comments are as follows.			Statistical sample sizes were unfavourably small.			reprodmed3020015_perova	0
+The entire set of experiments needs to be replicated to demonstrate reproducibility before it can be considered to publish these results.			demonstrate that PGRMC1 abundance levels are higher in proliferative phase versus secretory phase endometrial patient biopsies (fig.1).			reprodmed3020015_perova	0
+This is a critical point because the very title claims that PGRMC1 downregulation is required for decidualisation.			Author Response We appreciate your review of our revised manuscript, constructive comments, and suggestions, which have helped us to improve the manuscript.			reprodmed3020015_perova	0
+Unfortunately, the authors have been unable to demonstrate the direct involvement of PGRMC1 downregulation in decidualization.			And please provide basic information of the patient.			reprodmed3020015_perova	0
+The manuscript has been modified accordingly.			The salient point concerning the ubiquitination events is that they are correlated for other proteins with proteasome-mediated degradation.			reprodmed3020015_perova	0
+In fact, proteasomal degradation could be much more important that miRNA reduction of the transcript in the reduction of PGRMC1 levels during decidualization.			Suggest observations that may be able to discriminate between different possibilities.			reprodmed3020015_perova	0
+It has improved the readability, clarity, and quality of our manuscript.			Thus, the refined GGMs provide better local quasi-geoid results.			rs14061437_makarova	0
+Response to Reviewer 1 Comments: We thank you for your constructive and detailed comments.			It has improved the readability, clarity, and quality of our manuscript.			rs14061437_makarova	0
+If you have any information, please don’t hesitate to let us know.			Please comment on that question quantitatifely and qualitatively.			rs14061437_makarova	0
+The point-by-point response can be found from attachment.			Models Max Min Mean STD EIGEN-6C4 1.007 -1.696 0.048 0.187 GECO 1.579 -1.703 0.041 0.223 SGG-UGM-1 1.003 -1.671 0.052 0.194 SGG-UGM-2 1.003 -1.704 0.051 0.191 XGM2016 1.016 -1.757 -0.020 0.214 XGM2019 1.705 -1.737 0.081 0.213 Point 6: Your method for combining the satellite-only GFM with EGM2008 (sec.			rs14061437_makarova	0
+Email: zcli@cumtb.edu.cn 9 4 National Geomatics Center of China, Beijing 100830, China; zhangpeng@ngcc.cn; ys@ngcc.cn 10 * Correspondence: zcli@cumtb.edu.cn;Tel.			Author Response We thank you for your constructive and detailed comments.			rs14061437_makarova	0
+Introduction 33 With the emergence of Global Navigation Satellite System (GNSS), users can obtain 34 consistent ellipsoidal height at global scale.			Please comment on that question quantitatifely and qualitatively.			rs14061437_makarova	0
+3.3) seems quite simple for me and probably not optimal.			I have a few minor points (marked in the attached PDF) and two general points, which I like you to address.			rs14061437_makarova	0
+Importantly, even though all local height datums are re- 42 lated to the MSL, the vertical offsets between them may be up to 2 m at global scale [1].			The point-by-point response can be found from attachment, please see attachment.			rs14061437_makarova	0
+The point-by-point response can be found from attachment.			The ellipsoid height relative to a given geo- 35 centric ellipsoid can be obtained quickly and accurately by using GNSS.			rs14061437_makarova	0
+I have a few minor points (marked in the attached PDF) and two general points, which I like you to address.			Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach?			rs14061437_perova	0
+I have a few minor points (marked in the attached PDF) and two general points, which I like you to address.			Author Response We thank you for your constructive and detailed comments.			rs14061437_perova	0
+Why do you think, that a sharp cut-off SH degree between the satellite-only GFM and EGM2008 is better than a rigorous combination by applying some sophisticated weighting approach?			Dear authors, thank you for this interesting and well written study.			rs14061437_perova	0
+Email: zcli@cumtb.edu.cn 9 4 National Geomatics Center of China, Beijing 100830, China; zhangpeng@ngcc.cn; ys@ngcc.cn 10 * Correspondence: zcli@cumtb.edu.cn;Tel.			The goal of this paper is to derive 13 the geopotential value for the Chinese height datum (CNHD) in order to realize the height datum 14 unification in China.			rs14061437_perova	0
+29 Keywords: Chinese height datum; GRACE/GOCE; residual terrain model; spectral expansion ap- 30 proach; height datum geopotential 31 32 1.			Point 5: What is the main difference (except of the terrestrial data over China) to other combined GFMs, such as XGM2016 or XGM2019?			rs14061437_perova	0
+3.3) seems quite simple for me and probably not optimal.			Author Response We thank you for your constructive and detailed comments.			rs14061437_perova	0
+We have added descriptions and contents in the revised manuscript.			8 3 China University of Mining and Technology-Beijing, Beijing 100083, China.			rs14061437_perova	0
+If you have any information, please don’t hesitate to let us know.			However, the 36 ellipsoidal height is not related to the Earth's gravity field.			rs14061437_perova	0
+3.3) seems quite simple for me and probably not optimal.			: +86-0278-6752107 11 Abstract: A unified height datum is essential for global geographic information resource construc- 12 tion, ecological environment protection and scientific research.			rs14061437_perova	0
+However, I would like to make a few comments.			- I suggest presenting the paper's aim as the last idea in the introduction to make the manuscript easier to read.			s22072682_makarova	0
+Comments 4 I found some minor English errors throughout the paper.			In this respect, I find the work interesting, well-structured, and with promising results.			s22072682_makarova	0
+1 Response to Reviewer 1 Comments Comments 1 Please add a table with patients' baseline characteristics, including BMI and smoking status.			Comments 5 - I recommend deleting graphs 5, 8, and 11 because graphs 7, 10, and 13 show the same information together with lines obtained by analyzing the structural change of the time series.			s22072682_makarova	0
+1 Response to Reviewer 2 Comments Comments 1 -The paper's aim should be the same in the abstract and the introduction.			Comments 4 - In figure 3, the indication of the test time appears in the reverse direction.			s22072682_makarova	0
+Subject BMI Smoking history A 23.2 Non-smoker B 19.8 Non-smoker C 19.5 Non-smoker D 19.7 Non-smoker E 21.5 Non-smoker F 22.8 Non-smoker G 20 Non-smoker H 24.9 Non-smoker I 18.2 Non-smoker J 21.8 Non-smoker K 18.7 Non-smoker L 21.4 Non-smoker M 21.4 Non-smoker N 18.2 Non-smoker O 18.5 Non-smoker P 19.8 Non-smoker Q 20.5 Non-smoker mean 21 - SD 1.9 - Comments 2 Did authors estimate sample size?			However, I would like to make a few comments.			s22072682_makarova	0
+In this paper, using a maximal exercise test on a cycle ergometer, it is proposed to determine the anaerobic threshold using the strucchange analysis of the time series and compare it with the V slope method and the respiratory and metabolic indexes proposed by Wasserman.			"Dear Authors: I appreciate the possibility to review the paper entitled ""Exploration of an inflection point of ventilation parameters with anaerobic threshold using strucchange"" by the authors Aida et al."			s22072682_makarova	0
+Response 3: I described baseline load of the participants.			"Dear Authors: I appreciate the possibility to review the paper entitled ""Exploration of an inflection point of ventilation parameters with anaerobic threshold using strucchange"" by the authors Aida et al."			s22072682_makarova	0
+1 Response to Reviewer 1 Comments Comments 1 Please add a table with patients' baseline characteristics, including BMI and smoking status.			Comments 4 I found some minor English errors throughout the paper.			s22072682_makarova	0
+Response 2: I redraw Figures1-2 and described them as an adaptation of the original publication.			1 Response to Reviewer 1 Comments Comments 1 Please add a table with patients' baseline characteristics, including BMI and smoking status.			s22072682_makarova	0
+We are hopeful you find our responses/edits to your concerns (expressed below) sufficient.			Author Response Dear Reviewer 1, We are sending you the second version of our paper.			su14020860_makarova	0
+Author Response Dear Reviewer 3, Thank you for your kind comments.			We have tried to address all of your comments and take action on as many as we could.			su14020860_makarova	0
+Please, notice that our responses are provided in bold after your comments/suggestions.			We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			su14020860_makarova	0
+Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			su14020860_makarova	0
+We have tried to address all of your comments and take action on as many as we could.			While the paper seems interesting I have several concerns that I would like to share with the authors:			su14020860_makarova	0
+I am not convinced on your explanation on why committees would affect CSR.			We have tried to address all of your comments and take action on as many as we could.			su14020860_makarova	0
+Please, notice that our responses are provided in bold after your comments/suggestions.			Now, we would firstly like to thank you for your efforts in revising our study and we hope that the arguments and changes are satisfactory.			su14020860_makarova	0
+Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			However, I accept this article only for the moderating variable the authors have tested.			su14020860_makarova	0
+The paper lacks of robustness checks Authors:			While the paper seems interesting I have several concerns that I would like to share with the authors: The introduction lacks of motivation.			su14020860_makarova	0
+The article is methodologically OK and well written.			We have tried to address all of your comments and take action on as many as we could.			su14020860_perova	0
+We have tried to address all of your comments and take action on as many as we could.			The reader needs to understand why your research question is important.			su14020860_perova	0
+However, in general, we see many articles like this in the area of social and environmental reporting.			Indeed, I believe you have some omitted control variables which may cause an endogeneity issue and drive your findings.			su14020860_perova	0
+The paper studies the relationship between remuneration committees and CSR as well as the moderating role of independent directors.			While the paper seems interesting I have several concerns that I would like to share with the authors: The introduction lacks of motivation.			su14020860_perova	0
+We are hopeful you find our responses/edits to your concerns (expressed below) sufficient.			Author Response Dear Reviewer 3, Thank you for your kind comments.			su14020860_perova	0
+We appreciate your feedback and the time and care you spent with regards to reading our manuscript.			This study aims to explore whether the existence of remuneration committees tend to disclose more corporate social responsibility (CSR) information.			su14020860_perova	0
+Additionally, we have also introduced in the control variable paragraph more information to provide justifications on why those control variables are considered in this study.			Your empirical model needs to be reconsidered.			su14020860_perova	0
+We are hopeful you find our responses/edits to your concerns (expressed below) sufficient.			Author Response Dear Reviewer 3, Thank you for your kind comments.			su14020860_perova	0
+The conclusion needs to be rewritten to highlight the practical contributions of the paper Authors:			Having addressed all your proposals for change and improvement, we await your decision in the best interests of Sustainability.			su14020860_perova	0
+One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc.			On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118).			su14052976_makarova	0
+The line-up of below figures in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers.			"The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial."			su14052976_makarova	0
+One wonders whether the notion of transformative change really warrant this evolutionary perspective.			"The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial."			su14052976_makarova	0
+2, 3 and 4 above) in our manuscript (with two new figures compared to the original manuscript) has deliberately been chosen in response to a number of comments from reviewers.			8 Reviewer 1: A second point is the limited elaboration of the social contract, including the natural aspect.			su14052976_makarova	0
+Such an approach could help to better see the authors' contributions and the difference of this contribution to what has already been discussed about public governance, a debate that is nowadays quite robust.			Achieving this requires and involves a rebalancing of society and new imaginaries (as master signifiers), such as the well-being economy and a Natural Social Contract, as important new orientations.			su14052976_makarova	0
+Much of this, moreover, is action-oriented, pointing out major carriers and direction of change.			We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use.			su14052976_makarova	0
+In the given circumstances, and for the given topic, this seems very appropriate.			What could be helpful is, in your argumentation, briefly incorporate the genealogy of your key concepts, while you work with them, thus showing more clearly how you combine and transform them.			su14052976_makarova	0
+20 Reviewer 3: The paper presents quite an interesting discussion.			8 Reviewer 1: A second point is the limited elaboration of the social contract, including the natural aspect.			su14052976_makarova	0
+3 Editor: First of all, we would like to underline that it is ok to be normative.			The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			su14052976_makarova	0
+In a nutshell, to quote, the NSC “involves the fine-tuning of top-down policy and visions with important bottom-up processes (…), and takes place in a hybrid sphere, in which there is room for collaboration between governments, businesses, knowledge institutions and civil society, among others, characterized by Penta-helix models and based on multiple value creation”.			Comment Response by authors 1 Reviewer 1: “This paper has much improved, also because its scope is clearer.			su14052976_makarova	0
+A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement.			Our line of argumentation has been revised, and examples have been reduced to examples.			su14052976_makarova	0
+This would take care of the remarks about case studies and examples, and avoid the impression that there's several manifestos, or a manifesto immediately derived from one empirical situation.			The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary.			su14052976_makarova	0
+We have included a reference to the anxiety and (absent) feelings of control and competence.			10 Reviewer 1: Fourth, the section on co-evolution warrants serious re-thinking.			su14052976_makarova	0
+In particular, it is suggested to start from a systematic review on the studies that bring together co-evolutionary governance and the issue of sustainability, discussing what is the scientific advance of the proposition put forward here and how it enriches the current debate on these issues.			18 Reviewer 2: 5) In terms of the debate on public governance it would be important to insert the discussion of co-evolutionary governance not only ideally, but also exploring the differences between this theoretical proposition and other forms of governance already discussed in the literature as collaborative governance (Ansell and Gash, 2015), for example.			su14052976_makarova	0
+In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI).			"19 Reviewer 2: 6) Empirical cases are very relevant, but are placed as ""mere examples"" without delving into the challenges and difficulties lived in practice."			su14052976_makarova	0
+It is important that the authors give more emphasis on the scientific evidences of what they defend and anchor their argumentation in such evidences, whether theoretical or empirical.			Section 2 on “Transformation pleas and approaches for achieving this” has been substantially revised and we discuss three models for working towards transitions: collaborative governance, evolutionary steering and adaptive governance.			su14052976_makarova	0
+This paper is openly normative in the call for the creation of a Natural Social Contract and for transformation-oriented forms of governance, but the governance approaches advocated have been tried and tested, yielding insights for future use.			"The argument could certainly be presented more succinctly, and there is a need for a light language wash (""with the?"			su14052976_makarova	0
+The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary.			Because of this collaborative, polycentric approach, the papers draw on Evolutionary Governance Theory, and develops a perspective of “co-evolutionary steering of interrelated transitions”.			su14052976_makarova	0
+What could be helpful is, in your argumentation, briefly incorporate the genealogy of your key concepts, while you work with them, thus showing more clearly how you combine and transform them.			It is important that the authors also go into the challenges, the difficulties, the dilemmas in these debates.			su14052976_makarova	0
+And how then is the aspect of nature taken on board?			The authors cite several debates/definitions/approaches, but do not position themselves in relation to them, making clear what their theoretical contribution.			su14052976_makarova	0
+!” (France in anger) was a popular facebook site.			In this sense, it is not clear the relationship between the propositions presented by the authors in section 4 and the empirical or even theoretical evidences presented in the article.			su14052976_makarova	0
+All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			EGT understands governance as radically evolutionary and is mindful of limitations of any form of governance.			su14052976_makarova	0
+I suggest that the authors present the debate and the different definitions and then present the notion of GE, relating it to the TSEI.” Changes have been made accordingly.			In a nutshell, to quote, the NSC “involves the fine-tuning of top-down policy and visions with important bottom-up processes (…), and takes place in a hybrid sphere, in which there is room for collaboration between governments, businesses, knowledge institutions and civil society, among others, characterized by Penta-helix models and based on multiple value creation”.			su14052976_makarova	0
+To mention just one aspect, the text makes little or no reference to the differences and nuances between the realities and contexts of the countries of the North and the South, nor to the relationship between sustainability and social inequalities or between sustainability and democracy.			One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc.			su14052976_perova	0
+Format of figures and tables has been improved where appropriate.			With our major revision we believe we have addressed these comments, which resulted in a substantially improved paper.			su14052976_perova	0
+7 Reviewer 1: First, my main problem is that the paper reads as a compilation of manifestos, rather than one argumentative thread.			All three deal with complexity, normativity and uncertainty in an action-oriented way and have mechanisms of self-correction and adaptation which is necessary for transformations which cannot be planned and implemented from the top, but which must find solutions to demands for participation, fairness in outcomes, unintended effects in order to remain legitimate and secure continued support from societal stakeholders.			su14052976_perova	0
+Comment Response by authors 1 Reviewer 1: “This paper has much improved, also because its scope is clearer.			A social contract, as explained, consists of a consensual, value-based deal underpinning an institutional governance arrangement.			su14052976_perova	0
+Cover letter to Journal ‘Sustainability’ Special Issue: Environmental Policy and Governance: Evolutionary Perspectives Subject: Minor revisions / point-to-point response to reviewers’ second round of response Date: 14 February 2022 Manuscript ID: sustainability-1512113 Manuscript title: Transition policy after corona: The importance of a Natural Social Contract and co-evolutionary governance No.			16 Reviewer 2: 3) Make clearer and more explicit how the article meets the proposed problematic and responds to the gaps found in the debate, including raising the limits of such proposition.			su14052976_perova	0
+"We agree that the ""normative-prescriptive"" tone and use of ""should be"" was excessive."			I suggest to improve the interface between the propositions presented and what comes before in the text.			su14052976_perova	0
+11 Reviewer 1: Following on the latter, my final point concerns the relation between transformative change and NSC.			For more information see section 2 on page 3-10.			su14052976_perova	0
+The case at local and regional levels are well developed, but the discussion at international level seems poor, maybe the authors could exclude this mentioned focus o the Introduction I'll suggest a format review in the figures and their quotations, as well as the box and the table.			This would take care of the remarks about case studies and examples, and avoid the impression that there's several manifestos, or a manifesto immediately derived from one empirical situation.			su14052976_perova	0
+I suggest to improve the interface between the propositions presented and what comes before in the text.			3 Editor: First of all, we would like to underline that it is ok to be normative.			su14052976_perova	0
+In the section, NSC is presented as a ‘master signifier’ (345) and the orientation of governance actors (536) towards the ‘well-being economy’ (Table 1); NSC is also associated with alternative business models (443) and integral and robust forms of transition (554-581); NSC is said to need horizonal governance as well as Transformative Social-Ecological Innovation (TSEI).			The paper presents quite an interesting discussion.			su14052976_perova	0
+Indeed, it is partly a manifesto, but it also offers an action perspective in addition to an analytical perspective.			For more information see section 2 on page 3-10.			su14052976_perova	0
+I am still puzzled about the selection aspect of EGT, notably the specific roles of metaphors, open concepts and master signifiers play here (as objects and agents of selection) - particularly now NCS has been presented as a master signifier.			The paper’s aspiration is clear and laudable; yet its elaboration is wanting.			su14052976_perova	0
+"The theoretical debates and the literature on ""social contract"", ""public governance"" and ""sustainability"" are vast and controversial."			Two, the nature and form of evolutionary variation, selection and dynamics remain unclear.			su14052976_perova	0
+"15 Reviewer 2: 2) Remove from the problematization the ""normative-prescriptive"" tone starting from questions and nuisances that justify the study and from gaps in the literature."			We want to note that the governance approaches advocated have been tried and tested, yielding insights for future use.			su14052976_perova	0
+Comment Response by authors 1 Reviewer 1: “This paper has much improved, also because its scope is clearer.			The line-up moves from high abstraction level (figure 2), towards a tool for identifying options for co-evolutionary steering (figure 3), to an example of leverage points for a specific transition path (figure 4).			su14052976_perova	0
+Obviously, positive and normative approaches to support actionable agendas, like ours, do not escape the yardstick of scientific evidence.			The case at local and regional levels are well developed, but the discussion at international level seems poor, maybe the authors could exclude this mentioned focus o the Introduction I'll suggest a format review in the figures and their quotations, as well as the box and the table.			su14052976_perova	0
+Figure 2: Possible systemic leverage points for a societal transformation towards a Natural Social Contract, through transformative governance based on co-evolution across several interrelated dimensions (this figure is a synthesis of table 3.4 in Huntjens, 2021) Figure 3: The Intervention Flower as a tool for identifying options for co-evolutionary steering through connecting actor-coalitions and interdependent systemic leverage points.			I suggest to improve the interface between the propositions presented and what comes before in the text, specialy linking with the contributions and limites of the proposal to face this challenges.			su14052976_perova	0
+See section with new title: “Transformation pleas and governance approaches for achieving this”.			One of the reviewers speaks of a manifesto, and I would agree, but see it as a positive.			su14052976_perova	0
+On the other, the paper also refers to vested (eg ‘carbon’) interests (38, 619), strong voters’ resistance (574), policy holdups (507) and opposition from disadvantaged groups (118).			The question of how the NSC can help to meet and overcome such resistances has been addressed by diving into governance approaches that are capable of dealing with complexity, ambiguity and uncertainty, and with particular attention for multiple value creation and the mutual gains approach (e.g.			su14052976_perova	0
+Author Response Please see the attachment Author Response File: Author Response.docx			One the one hand, the paper trumpets collaboration, joint principles and responsibilities, mutual trust, etc.			su14052976_perova	0
+Two cases serve as quick illustrations, loosely underwriting some key statements.			The paper’s general, theoretical story is very broad and ambitious, using an extensive, abstract and complex vocabulary.			su14052976_perova	0
+Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information.			I still encourage the authors to have a deep reflection on the legal and population acceptance of their proposal, but this can be the goal of another paper.			su14052981_makarova	0
+The authors than write that these services have a limitation in that cannot predict and prevent transmission of threats such as COVID.			As a result, the critical point of being identified before an infectious disease spreads was missed.			su14052981_makarova	0
+Section 2.3 has the same title as Section 2.2.			I think that is both a big oversight, and an opportunity for critics.			su14052981_makarova	0
+I will give a clearer example from p.15, line 345: Second, the ( Figure			Regarding the content, the motivation is clearer in the abstract, and this clarifies the contribution as well.			su14052981_makarova	0
+Table 4 presents smart city projects of several different countries.			line 85) The reference confirmed that the error occurred when the formatting was moved.			su14052981_makarova	0
+The authors do not explain how the services they propose could be integrated with existing ones (e.g.			Author Response Please see the attachment Author Response File: Author Response.pdf			su14052981_makarova	0
+Several different plans are described, which is useless information for the reader.			I still encourage the authors to have a deep reflection on the legal and population acceptance of their proposal, but this can be the goal of another paper.			su14052981_makarova	0
+The authors acknowledge that not addressing these is a limitation of their research.			Line 320: according to the introduction the COVID-19 pandemic started in 2019 and here you state 2020.			su14052981_makarova	0
+I was not conviced by the authors' response regarding my suggestion for putting 3.1 into Section 2, but this was a quite minor comment anyway, and there is not only one good way to structure a text, the paper is also understandable with the structure proposed by the authors.			Regarding the content, the motivation is clearer in the abstract, and this clarifies the contribution as well.			su14052981_makarova	0
+First, an epidemiological surveyor should decide whether additional collection of personal information is needed.			I think many cities and countries will not be comfortable with this approach.			su14052981_makarova	0
+However some parts still appear unnecessary to me.			In this source you can find a whole list of smart city technologies http://www.sinfonia- smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work.			su14052981_makarova	0
+Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors.			Regarding the content, the motivation is clearer in the abstract, and this clarifies the contribution as well.			su14052981_makarova	0
+Wastewater surveillance for population-wide Covid-19: the present and future.			For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source.			su14052981_makarova	0
+Thank you again for your supportive review to improve the manuscript.			Author Response Please see the attachment Author Response File: Author Response.pdf			su14052981_makarova	0
+Once infected people have been identified, they are supposedly in quarantine.			I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable.			su14052981_makarova	0
+In Smart Cities for Technological and Social Innovation (pp.			I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors.			su14052981_makarova	0
+The authors argue that the smart city services failed to be flexible enough to predict and fight the pandemic.			should appear as Second, the Figure 4 scenario assumed by this study is uncertain.			su14052981_makarova	0
+The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model.			I believe that explaining standard constraints (due to the multiplicity of service providers) and coupling between services in the introduction to motivate the paper would be valuable.			su14052981_makarova	0
+Thank you again for your supportive review to improve the manuscript.			In summary, my suggestions are to focus the paper on the Korean context, in an in-depth case study approach where authors detail more their contribution, clearly illustrate what the problem is and how their contribution helps solving it.			su14052981_makarova	0
+The contribution is a proposition of services such as screening people with thermal cameras and tracking infected people through CCTV.			Smart City and COVID Webb, W., & Toh, C. K. (2020).			su14052981_makarova	0
+The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper.			I will give a clearer example from p.15, line 345: Second, the ( Figure			su14052981_makarova	0
+The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.			Wastewater surveillance for population-wide Covid-19: the present and future.			su14052981_makarova	0
+Current Opinion in Environmental Science & Health, 17, 14-20.			- “This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.” #4.			su14052981_makarova	0
+Additionally, I've included a few other pieces on COVID and smart cities.			“Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense?			su14052981_makarova	0
+Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.			Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography.			su14052981_makarova	0
+Unfortunately, they are enough for me to recommend to reject the paper, as I consider that submitting a report of scientific quality in this form is unacceptable.			The authors than write that these services have a limitation in that cannot predict and prevent transmission of threats such as COVID.			su14052981_makarova	0
+Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors.			“Additionally, smart city services couldn`t respond in-time series” (is this time series in the statistical sense?			su14052981_makarova	0
+The authors made substantial efforts to take my comments into account, and I commend them for that.			There are also many in-text reference errors with tables, which makes it hard to follow given the high number of tables (13!).			su14052981_makarova	0
+Wastewater surveillance for population-wide Covid-19: the present and future.			Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			su14052981_makarova	0
+This may be because of the error message 3 in sourcing in the document I received.			Section 3.1 gives an overview of the COVID-19 and smart city systems situation in South Korea, but is not part of the novel contribution by the authors.			su14052981_makarova	0
+I still encourage the authors to have a deep reflection on the legal and population acceptance of their proposal, but this can be the goal of another paper.			Once infected people have been identified, they are supposedly in quarantine.			su14052981_makarova	0
+COVID-19 Scenario: Outbreak of COVID-19 in Korea ) scenario assumed by this study is uncertain.			However, despite the good improvements, I have some remaining concerns.			su14052981_makarova	0
+Thank you again for your supportive review to improve the manuscript.			I was not conviced by the authors' response regarding my suggestion for putting 3.1 into Section 2, but this was a quite minor comment anyway, and there is not only one good way to structure a text, the paper is also understandable with the structure proposed by the authors.			su14052981_makarova	0
+In Smart Cities, Green Technologies, and Intelligent Transport Systems (pp.			Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			su14052981_makarova	0
+Wastewater and public health: the potential of wastewater surveillance for monitoring COVID-19.			The legal and ethical aspects are still too much overlooked in my opinion, but I appreciate that the authors added a note on the legal framework of Korea.			su14052981_makarova	0
+Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information.			There are referencing issues with sections as well (e.g.			su14052981_makarova	0
+The abstract is very confusingly written and needs a lot of editing.			This is a very important point, since it changes what is the contribution of the paper.			su14052981_makarova	0
+There are some phrasing issues and typos throughout the paper.			As a result, the critical point of being identified before an infectious disease spreads was missed.			su14052981_makarova	0
+Current Opinion in Environmental Science & Health, 17, 14-20.			I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned.			su14052981_makarova	0
+The Impact of COVID-19 Experience on Smart City and Future Mobility.			Indeed, if my understanding is correct, they are the factors that cause the limitations of smart city services the authors want to address.			su14052981_perova	0
+I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations.			In addition, errors in the sentences have been reviewed and corrected.			su14052981_perova	0
+I stand by my position that the feasibility/ethical aspects should be discussed, at least from a social acceptance or a legal point of view, as the reader is left skeptical about whether it can actually be done with the current explanations.			1 Dear Reviewer #1, I am grateful for your suggestions and kind commendation.			su14052981_perova	0
+In Smart Cities for Technological and Social Innovation (pp.			Once infected people have been identified, they are supposedly in quarantine.			su14052981_perova	0
+Sharifi, A., Khavarian-Garmsir, A. R., & Kummitha, R. K. R. (2021).			First, I have a series of concerns regarding the form.			su14052981_perova	0
+It is also hard to read, as it contains more tabular content than text.			Please check especially the conclusion, this is hardly readable.			su14052981_perova	0
+Smart City and COVID Webb, W., & Toh, C. K. (2020).			In other terms, what can we use from the authors' paper to achieve a more flexible smart city service structure?			su14052981_perova	0
+The authors bring up interesting legal considerations in their response to my comments, it would be nice to have them in the paper to show the reader under which conditions the authors' solution can be used.			I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned.			su14052981_perova	0
+The legal and ethical aspects are still too much overlooked in my opinion, but I appreciate that the authors added a note on the legal framework of Korea.			It is important to operate the monitoring system for sewage and wastewater for population level assessment of infection, which is considered to be the most important source of infection.			su14052981_perova	0
+We sincerely apologize for the basic mistake.			Lines 352-354: here we have two times Figure 2.			su14052981_perova	0
+I will give a clearer example from p.15, line 345: Second, the ( Figure 4.			The legal and ethical aspects are still too much overlooked in my opinion, but I appreciate that the authors added a note on the legal framework of Korea.			su14052981_perova	0
+Section 2.3 has the same title as Section 2.2.			It is also hard to read, as it contains more tabular content than text.			su14052981_perova	0
+Thank you again for your supportive review to improve the manuscript.			In addition, we will use a more advanced paper by reflecting it in the ongoing research.			su14052981_perova	0
+The authors argue that this would be precious to predict and fight against a pandemic, and as such they propose a smart city structure model.			1) Confusing or awkward wording in the abstract:			su14052981_perova	0
+I have checked the online source and it mentions other aspects such as treatment and isolation for the recognition phase, which have not been picked up by the authors.			1 Dear Reviewer #2, I am grateful for your suggestions regarding my paper, and my answers to your comments are provided below.			su14052981_perova	0
+In addressing the comments, I have been able to improve the clarity and content of the manuscript vastly.			Thank you once again for your pertinent suggestions and feedback.			su14052981_perova	0
+Several different plans are described, which is useless information for the reader.			For technical reasons (most smart cities have less capability and are less developed than Korean ones), but especially for legal and normative reasons (concerns about surveillance, governance, cybersecurity, etc.)			su14052981_perova	0
+The literature review is very long (it represents half of the paper) and can be significantly shortened as it contains few information that is actually useful to the rest of the paper.			In addition, errors in the sentences have been reviewed and corrected.			su14052981_perova	0
+I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned.			Please feel free to add them back if you feel they should be included.			su14052981_perova	0
+This is a very important point, since it changes what is the contribution of the paper.			Please check especially the conclusion, this is hardly readable.			su14052981_perova	0
+I have made major revisions to the paper based on your inputs, as mentioned below.			(pg3) Tables 1 is interesting, but it’s not clear the source of the data.			su14052981_perova	0
+Although there are still many things incomplete, we did our best to reflect the opinions of the reviewers.			1) Confusing or awkward wording in the abstract:			su14052981_perova	0
+The authors made substantial efforts to take my comments into account, and I commend them for that.			For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source.			su14052981_perova	0
+I thus find it now less essential to discuss the practical aspects of service adding and removing.			However, I am lacking the contribution on the more general issue of integrating and removing services in an existing smart city infrastructure.			su14052981_perova	0
+In this source you can find a whole list of smart city technologies http://www.sinfonia- smartcities.eu/en/resources/d21--swot-analysis-report-of-the-refined-conceptbaseline Might be of interest for your work.			[15], [22], [25]) it is impossible to retrieve the source.			su14052981_perova	0
+Thank you once again for your pertinent suggestions and feedback.			The authors could remove the table and mention 1-2 standards relevant for pandemics and make the same point.			su14052981_perova	0
+(pg2) Does the use of police/telecom monitoring of patients translate out of the Korean context?			However, it misses a critical point of COVID.			su14052981_perova	0
+It is well-known that many standards exist and are not used in every service, which causes integration issues.			Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information.			su14052981_perova	0
+The abstract is very confusingly written and needs a lot of editing.			Regarding the references, my point was that there are many references *with formatting mistakes in them*, and not that there are too many references in the bibliography.			su14052981_perova	0
+Section 3, named “Main discussion”, should be renamed to be more explicit regarding what the reader can expect to find.			It is well-known that many standards exist and are not used in every service, which causes integration issues.			su14052981_perova	0
+My last concern is the ethical aspects of the proposed services.			Third, smart city services should only be used to cope with pandemic situations: 18 the healthcare-system data consists of personal information.			su14052981_perova	0
+I have made major revisions to the paper based on your inputs, as mentioned below.			Thank you once again for your pertinent suggestions and feedback.			su14052981_perova	0
+Additionally, I've included a few other pieces on COVID and smart cities.			Do the authors propose to track everyone and then go back to the records of infected people to retrieve their paths and contacts?			su14052981_perova	0
+For many other sources, the local access path of the file is provided, which again prevents the reader from retrieving the source.			- “This study proposes a method for timely and sequential responses, through a flexible combination of the healthcare system and smart city services by envisioning a scenario that sequentially grafts the current status of COVID-19 in Korea.” #4.			su14052981_perova	0
+Section 2 would thus have a part on smart city services, a part on COVID-19 response plans, and then the current content of 3.1 which deals with the relationship between the two parts of the current Section 2.			In the introduction, I could not understand the motivation of the paper.			su14052981_perova	0
+Also, references to figures seem to include the full caption and cause line breaks instead of the figure number only.			I think that is both a big oversight, and an opportunity for critics.			su14052981_perova	0
+However some parts still appear unnecessary to me.			I think it synthesizes some important and useful literatures, and has some useful points to make.			su14052981_perova	0
+I'd argue that one of the areas in which Smart City applications (in particular) and urban automation (in general) have been most important during the COVID pandemic - the monitoring of sewage and wastewater for population level assessment of infection - isn't even mentioned.			Thank you again for your supportive review to improve the manuscript.			su14052981_perova	0
+Therefore, smart city services respond- 19 ing COVID-19 must exist as a flexible.”			Also, references to figures seem to include the full caption and cause line breaks instead of the figure number only.			su14052981_perova	0
+We sincerely apologize for repeating basic mistakes.			should appear as Second, the Figure 4 scenario assumed by this study is uncertain.			su14052981_perova	0
+You have some values that are not included in this interval - maybe because of the outliers from the figures above that I recommend to exclude.			However, there is no in-depth discussion or recommendations that follow.			su14095590_perova	0
+The authors presented and analyzed the variables/areas of certification with many models, it is interesting and is a basis for analysis in the next section.			Literature review: There is still work to do discussing the five areas, not just relating them to each dimension of the Triple Bottom Line of the sustainability concept.			su14095590_perova	0
+Reviewer’s suggestion: This paper analyses the consistency and robustness of the B impact assessment model in measuring sustainability impacts in five areas of ?			What is the main result of the B assessment with the confirmatory analysis?			su14095590_perova	0
+Point 2: Further, the hypothesis does not seem to be written correctly.			The authors test a popular assessment tool using confirmatory factor analysis and finds that the model is weak.			su14095590_perova	0
+The method section has been improved (text in blue).			The authors should develop more theory than describe a model which has already existed.			su14095590_perova	0
+The Cronbach’s Alpha ‘selected’ just ‘environment’ area of the B Lab model.			Point 2: Are the research design, questions, hypotheses and methods clearly stated?			su14095590_perova	0
+Abstract: The main objective is to analyze the B Impact Assessment, verifying the robustness and consistency of the model to measure and improve the economic, social, and environmental impact of companies.			It’s their signature in the text, so I understand that they deserve this recognition.			su14095590_perova	0
+Point 3: The text has some improvements, but there are some points that still demand more attention.			Point 6: Results: The sentence from line 137 to line 138 isn’t clear when I saw Figure 6.			su14095590_perova	0
+Point 6: Results: The sentence from line 137 to line 138 isn’t clear when I saw Figure 6.			The authors answer the comments by saying that Figure 1 was done by them, it’s fair to provide this information.			su14095590_perova	0
+In fact, the discussion needs to be grounded in previous research findings in regard to CSR/sustainability indices/assessment tools/initiatives.			Overall the information presented it makes sense to me.			su14095590_perova	0
+Some authors were added, but for me, this section demands an improvement to enhance the quality of the text, and to turn it into a scientific paper.			It was not clear to me who is the author of Figure 1, the B Lab, or the authors of this paper?			su14095590_perova	0
+The manuscript has been revised to improve grammar and english language.			We have made all the minor corrections indicated.			su14095590_perova	0
+Point 3: Unfortunately, I cannot recommend publication unless major changes are made to the discussion section and possibly recommendations are added for future research and practice.			First, it should be stated prior to the methods section.			su14095590_perova	0
+The authors must add the source of that areas, and it is enough for the objective.			Overall the information presented it makes sense to me.			su14095590_perova	0
+For me, the answer to Hypothesis 1 isn’t clear.			1 Response to Reviewer 1 Comments Point 1: Thank you for giving the opportunity to read such an interesting paper.			su14095590_perova	0
+The section demands a literature review about the theme, as the relationship between the certificate with adding value, sustainability, and the ‘areas’ governance, workers, community, environment, and customers.			Line 199, L. 223) • Paragraph presented after Table 2 (page 7) is a repetitive content to what has been presented in the Table.			su14095590_perova	0
+- workers values greater then 40, community greater than 58, environment 45, customers 46 (the bullets), in Fig 3 - values with global score greater than 130, in Fig.			1 Response to Reviewer 1 Comments Point 1: Thank you for giving the opportunity to read such an interesting paper.			su14095590_perova	0
+For me, the answer to Hypothesis 1 isn’t clear.			The source of data from all companies in the sample isn’t clear, is B Corporation (site) or authors contact all these companies and did the survey?			su14095590_perova	0
+The authors answer the comments by saying that Figure 1 was done by them, it’s fair to provide this information.			1 Response to Reviewer 3 Comments (round 2) Point 1:			su14095590_perova	0
+The sentence in the Introduction is more complete than the main objective in the Abstract.			What is its contribution to the structure of a model using Confirmatory Analysis?			su14095590_perova	0
+• It is better to show the P-values in Table 3 rather than (***) although a note is indicated in this regard.			This is an interesting result, which could be more analyzed with other assessment models presented in the text, to say what the authors wrote at the end of the section.			su14095590_perova	0
+I suggest the explanation given in the reviewers’ response about the decrease of companies in the sample be available in the text (“The reduction in the number of companies was based on the analysis of the database and information collected from B Lab that allowed us to see that there was a change in the measurement model used in B Corp certification during the period January 2017 and March 2021.			The authors should develop more theory than describe a model which has already existed.			su14095590_perova	0
+The content in this section is exclusively dedicated to the Certification and the Corporation which offers it, there isn’t any scientific information there.			• It is better to show the P-values in Table 3 rather than (***) although a note is indicated in this regard.			su14095590_perova	0
+Point 2: Abstract: The content is well organized, but the results need some to attract the reader.			Point 2: Are the research design, questions, hypotheses and methods clearly stated?			su14095590_perova	0
+Response 5: We believe that the reviewers' suggestions have been very helpful in improving the manuscript.			For me, the answer to Hypothesis 1 isn’t clear.			su14095590_perova	0
+For me, lines 39 and 40 show the aim of the paper, but the idea is different from the ones presented in the abstract (“The main objective is to understand the B Impact Assessment, verifying its added value in the sustainability process and socio-economic development of the business sector”).			I suggest cutting the text from lines 90 to 103 because it’s quite an advertisement for the B Certificate.			su14095590_perova	0
+Overall, I believe that this study has a high potential to add to the literature on the relationship between gender diversity and firm environmental performance.			This is particularly relevant for countries vulnerable to shocks whether internal or external (Covid-19, war, conflicts, trade tensions, etc.).			su14127346_makarova	0
+Below we provide the details on how your comments and suggestions have been addressed.			In terms of policy, our findings suggest that promoting gender diversity cannot reach full potential, and may even lead to adverse results unless accompanied by measures that mitigate political and economic uncertainty.			su14127346_makarova	0
+This, in turn, discourages in-vestments in costly abatements technologies and environmental-friendly processes.			The rational for this result stems from the idea that uncertainty adversely affects the enabling environment of an economy.			su14127346_makarova	0
+In terms of policy, our findings suggest that promoting gender diversity cannot reach full potential, and may even lead to adverse results unless accompanied by measures that mitigate political and economic uncertainty.			In terms of policy, our finding suggests that promoting gender diversity on corporate boards should be accompanied by measures that reduce political and economic uncertainty.			su14127346_makarova	0
+This threshold value points to a standard and very important result in the literature.			These reduce their aversion to risk, which increases their confidence and willingness to invest.			su14127346_makarova	0
+These measures would create an enabling environment both for households and entrepreneurs.			This refers to the critical mass theory, by which the impact of gender diversity on the environmental performance of firms materializes only when a critical mass weight of females is realized in corporate boards (Konrad et al., 200839; Torchia et al., 201140; Schwartz-Ziv, 201741; among others).			su14127346_makarova	0
+Amended data is already available on the product.			As it stands, the paper offers little insight into the reasoning behind their hypothesis.			su14127346_makarova	0
+More gender-diverse corporate boards would not lead to better environmental performances of firms in countries with higher uncertainty (higher WUI score).			Please provide some, and position your work in that (brief) strand of literature.			su14127346_makarova	0
+This is a well-executed study with a solid research base.			4- Add a discussion of results section and relate your results with theories.			su14127346_makarova	0
+Similar problems might exist for a negative tobin's Q.			Similar problems stem from the following paragraphs discussing the results.			su14127346_makarova	0
+Ultimately, this would put pressure on firms’ behavior and worsen their environmental performances (Bloom, 20149; Atsu and Adams, 202137).			The only remark on the content refers to “Are the arguments and discussion of finding coherent, balanced and compelling?			su14127346_makarova	0
+It would be useful if the authors provided some speculative thoughts on the finding that the slope of the GenDiv variable is negative in highly uncertain countries on the firm's environmental performance (Figure 3 (c) red line).			The arguments above provides a rationale for the worsened environmental performances of firms in response to higher uncertainties, and explains the gap in effects for firms across countries of different uncertainty levels.			su14127346_makarova	0
+Similar problems might exist for a negative tobin's Q.			I only have a few minor concerns that I'd like the authors to clarify and address when revising.			su14127346_makarova	0
+Ultimately, this would put pressure on firms’ behavior and worsen their environmental performances (Bloom, 20149; Atsu and Adams, 202137).			We deeply regret the inconvenience caused due to this.			su14127346_makarova	0
+They contribute to anchoring economic agents’ expectations, and stabilize the economy.			They contribute to anchoring economic agents’ expectations, and stabilize the economy.			su14127346_makarova	0
+I would like to recommend its acceptance for publication.			I would like to recommend its acceptance for publication.			toxins14060409_makarova	0
+Among the three methods used, the upper bilateral floret injection (UBFI) maximizes DON accumulation in grain and significantly distinguishes DON content in different wheat genotypes, indicating that the method could be useful for breeding program to develop wheat cultivars with resistance to DON accumulation.			The paper describes a set of experiments carried out in 2020 2021 on isogenic lines of wheat using different methods of Fusarium inoculation in order to assess the effect of Fusarium inoculation on FHB disease index and DON accumulation.			toxins14060409_makarova	0
+Mycotoxin deoxynivalenol (DON) contamination of wheat and other grains has been a concern of food safety.			Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			toxins14060409_makarova	0
+Minimizing DON contamination in grains is a major goal of Fusarium Head Blight (FHB) research community.			Point 7: Fhb1- and Fhb1+ don’t appear in Table S1, so they can be removed from the footnote.			toxins14060409_makarova	0
+Mycotoxin deoxynivalenol (DON) contamination of wheat and other grains has been a concern of food safety.			Point 2: Line 142: “proportion of PPSD” should be “proportion of PSD or PPSD”.			toxins14060409_makarova	0
+Given that 2.4 chapter discusses data of supplementary figure 4 would suggest to make figure S4 as Figure 4 of the manuscript.			Point 2: To further clarify the infection methods a supplementary figure detailing graphically the modes of inoculation with figures would make the paper extremely useful for the community.			toxins14060409_makarova	0
+1 Response to Reviewer 2 Comments Point 1: I would suggest to avoid citing supplementary figures at the beginning of the chapter.			Among the three methods used, the upper bilateral floret injection (UBFI) maximizes DON accumulation in grain and significantly distinguishes DON content in different wheat genotypes, indicating that the method could be useful for breeding program to develop wheat cultivars with resistance to DON accumulation.			toxins14060409_makarova	0
+Given that 2.4 chapter discusses data of supplementary figure 4 would suggest to make figure S4 as Figure 4 of the manuscript.			Authors in this manuscript report that DON accumulation in wheat grain is affected by inoculation methods (infection points).			toxins14060409_makarova	0
+1 Response to Reviewer 2 Comments Point 1: I would suggest to avoid citing supplementary figures at the beginning of the chapter.			The paper describes a set of experiments carried out in 2020 2021 on isogenic lines of wheat using different methods of Fusarium inoculation in order to assess the effect of Fusarium inoculation on FHB disease index and DON accumulation.			toxins14060409_makarova	0
+While DON concentrations in grains are usually correlated with disease severity, high DON levels have been observed in asymptomatic kernels.			Point 7: Fhb1- and Fhb1+ don’t appear in Table S1, so they can be removed from the footnote.			toxins14060409_makarova	0
+Minimizing DON contamination in grains is a major goal of Fusarium Head Blight (FHB) research community.			1 Response to Reviewer 1 Comments Point 1: I would like to suggest authors label inoculation points in Figure 1, which could give readers a clear view of three different inoculation methods.			toxins14060409_makarova	0
+This may be true, but far more studies have confirmed this link than have refuted it.			The IFITM3 blot shows wide variability with no pattern from one hour to the next, and yet the conclusion is that there is no change in IFITM3 levels during this time period.			v11060548_makarova	0
+Figure 1 should be labeled as to what treatments are used in panels A-E. As it is now, the figure on its own cannot possibly be interpreted without the legend.			d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.			v11060548_makarova	0
+There are few clarifications that would strengthen the overall manuscript.			Additional minor comments on the figures include: a.			v11060548_makarova	0
+At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			Supplemental figures should be included in the main text.			v11060548_makarova	0
+There are few clarifications that would strengthen the overall manuscript.			d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.			v11060548_makarova	0
+At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.			v11060548_makarova	0
+Using super-resolution microscopy the authors show colocalization of IFITM3 with influenza viral nucleoprotein and early endosomal proteins.			c. Labels or zoomed out images from those presented in Figure 3 and 7 are needed to orient the reader to where within a cell the colocalization is observed.			v11060548_makarova	0
+If this is not acceptable we will make the changes in final revision.			IFITM3 protein increased significantly after longer infection times such as 24 h p.i.			v11060548_makarova	0
+Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			v11060548_makarova	0
+Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity.			Additional minor comments on the figures include: a.			v11060548_makarova	0
+This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species.			Additionally, a second SNP in the IFITM3 promoter has also been linked to severe flu.			v11060548_makarova	0
+At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			This does not appear to increase post infection as was shown previously.			v11060548_makarova	0
+They show that IFITM3 clusters within 1 h of influenza virus infection, prior to increases in the protein’s levels as induced by the infection.			qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.			v11060548_makarova	0
+These IFITM3 clusters were found to correlate with restriction of virus infection and to co-localize with incoming virus.			There are few clarifications that would strengthen the overall manuscript.			v11060548_makarova	0
+Lines 67-73 The inhibition of virus membrane fusion by IFITM3 has been attributed to the presence of a palmitoylated amphipathic helix within IFITM3 (Chesarino, EMBO Reports, 2017).			There are few clarifications that would strengthen the overall manuscript.			v11060548_makarova	0
+Using super-resolution microscopy the authors show colocalization of IFITM3 with influenza viral nucleoprotein and early endosomal proteins.			In addition, the movie does not provide any details on cellular features and a viewer is left wondering where the foci are coming from and going.			v11060548_perova	0
+The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining.			Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			v11060548_perova	0
+d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.			The largest concern is regarding the colocalization of IFITM3 and Rab11A.			v11060548_perova	0
+The NP stain in HSAEpCs is only shown to be cytoplasmic, inclusion of an uninfected control would demonstrate specificity of the staining.			For a review of these studies, see Zani, Current Clin Microbiol Reports, 2018, though the primary articles should be cited.			v11060548_perova	0
+Supplemental figures should be included in the main text.			This helix and neighboring palmitoylation sites are among the most highly conserved residues among IFITMs from all species.			v11060548_perova	0
+qPCR is acceptable, but an IFITM3 Western would be better, particularly since the antibody is readily available.			d. Figure 2B - Western blot examining levels of IFITM3 should include 24 hpi to correlate with the data presented in Figure 1.			v11060548_perova	0
+Additionally, the role of cholesterol in IFITM3’s mechanism of action has been largely disproven by the field, and reference 52 seems to have been misused in line 73.			Following the suggestion of this reviewer, we have removed the movie in the revised version.			v11060548_perova	0
+Figure 3A: Why is there so much IFITM3 present in the uninfected cells?			At later timepoints is the NP stain nuclear as expected, is this simply a delay in replication?			v11060548_perova	0
+Figure 4: Label the figure so that it can be interpreted independent of the legend.			Further, they observed that the IFITM3 clusters that form early in infection are at early endosomes and progress to late endosomes and lysosomes as infection proceeds, consistent with IFITM3 directing incoming virus to degradative compartments.			v11060548_perova	0
+Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			In addition, the movie does not provide any details on cellular features and a viewer is left wondering where the foci are coming from and going.			v11060548_perova	0
+Lines 227/228: This sentence is misleading as the references cited are not contradictory, nor are they unclear as to IFITM3 possessing antiviral activity.			Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.			v11060548_perova	0
+If this is not acceptable we will make the changes in final revision.			This study provides imaging of the antiviral protein IFITM3 at endogenous levels and at super-resolution in the context of influenza virus infection.			v11060548_perova	0
+Further, the studies which failed to find an association were generally performed in populations in which the SNP is almost non-existent.			Therefore the inclusion of the movie examining the colocalization of Rab11A GFP, IFITM3-SNAP, and labeled virion seems completely unnecessary.			v11060548_perova	0
+Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			Additional minor comments on the figures include: a.			v11060548_perova	0
+This may be true, but far more studies have confirmed this link than have refuted it.			Figure 5A/C: Label the figure so that it can be interpreted independent of the legend.			v11060548_perova	0
+Following the suggestion of this reviewer, we have removed the movie in the revised version.			Furthermore, the authors need to examine the literature more carefully, as previous studies have indeed shown that IFITM3 knockdown in A549 cells increases influenza virus infection, e.g., Lin, Cell Reports, 2013.			v11060548_perova	0
+Author Response Thank you very much for your support and help.			Authors may provide data in the supporting information file.			w14030367_perova	0
+The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water.			Please target the specific gap such as 2015-2021 etc Response 3:			w14030367_perova	0
+Techno-economic assessment of Minimal Liquid Discharge (MLD) treatment systems for saline wastewater (brine) management and treatment.			The reviewer suggests statistical analysis using a t-test (Raw water- BAC-Effluent and Raw water – BAC/UF-effluent).			w14030367_perova	0
+In the present form conclusion is very weird.			Response 7: Thank you very much for your kind reminder.			w14030367_perova	0
+In the present form conclusion is very weird.			I can see very well-defined experiments and produced good results.			w14030367_perova	0
+The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances.			Point 14: Please use one style for units such as m3/h or m3h-1 Please revise your paper accordingly since some issue occurs on several spots in the paper.			w14030367_perova	0
+Size exclusion is considered the primary removal mechanism for the UF.			The manuscript could be accepted for publication after a minor revision.			w14030367_perova	0
+Response 11: Thanks very much for this comment.			They also discussed their results in detail and put the difference in their results compared to the data with the literature.			w14030367_perova	0
+Point 5: Conclusion: Make it as one or two paragraphs.			Point 5: Line 53-55: “The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”.			w14030367_perova	0
+Two native English-speaking colleagues help us verify the manuscript.			Results seem to be related to a unique experiment.			w14030367_perova	0
+Response 5: Thanks very much for this comment.			Study and evaluation of the characteristics of saline wastewater (brine) produced by desalination and industrial plants.			w14030367_perova	0
+Response 6: Thanks very much for this comment.			Response to Reviewer 4 Comments Point 1: Authors should avoid abbreviations in the title and the abstract.			w14030367_perova	0
+Point 9: Please check the abbreviations of words throughout the article.			However, in the case of the PPCPs with a small molecular weight (typically < 600 Da), UF membranes also cannot effectively reject these PPCPs, but nanofiltration and reverse osmosis are able to remove these PPCPs based on the thin-film composite”.			w14030367_perova	0
+The article topic is intriguing and promising in the area.			Response 13: Thank you very much for your kind reminder.			w14030367_perova	0
+It was found that the BAC unit 25 played a key role in PPCPs removal and the UF unit also degraded them by the biomass on UF 26 membranes.			The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water.			w14030367_perova	0
+In the present form conclusion is very weird.			: +86 15765532017; E-mail: chnwangjinlong@163.com (J. Wang) 14 Abstract： Biological activated carbon (BAC) biofilter coupling ultrafiltration (UF) is a promising 15 process for the treatment of surface water contaminated by pharmaceutical and personal care prod- 16 ucts (PPCPs).			w14030367_perova	0
+The comparison of previous research with existing research findings was added in the final part of the results and discussion.			Response 9: Thanks very much for this comment.			w14030367_perova	0
+Response 8: Thanks very much for this comment.			“The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”.			w14030367_perova	0
+If possible, please add at least one paragraph.			Response 2: Thank you very much for your kind reminder.			w14030367_perova	0
+Please revise your paper accordingly since some issue occurs on several spots in the paper.			43 Citation: Wanga, Q.; Tanga, X.; Zenga, W.; Wangb, F.; Gongb, W.; Chena, J.; Wanga, J.; Lia, G.; Lianga, H. Pilot-scale biological activated carbon filtration-ultrafiltration (BAC-UF) system for removing pharmaceutical and personal care products (PPCPs) from surface wa- ter.			w14030367_perova	0
+Point 9: Point 6: Why the same trend of increase or decrease in graphs (Fig.			Sorry, but I don't see why UF can reject viruses and bacteria but not PPCPs.			w14030367_perova	0
+Point 9: Point 6: Why the same trend of increase or decrease in graphs (Fig.			Response 6: Thanks very much for this comment.			w14030367_perova	0
+Greetings, Editor thank you for providing me with the opportunity to review the article.			Please clarify, why ultrafiltration cannot remove the PPCPs since it can remove bacteria and viruses.			w14030367_perova	0
+Please also see the attached file for my corrections/comments.			Size exclusion is considered the primary removal mechanism for the UF.			w14030367_perova	0
+Two native English-speaking colleagues help us verify the manuscript.			The comparison of previous research with existing research findings was added in the final part of the results and discussion.			w14030367_perova	0
+Point 3: Introduction section must be written on more quality way, i.e., more up-to-date references addressed.			Generally, many PPCPs 41 flow through conventional water treatment processes with little degradation due to their 42 persistency or/and the continuous introduction [5].			w14030367_perova	0
+The concentrations and diversities complicate the associated detec- 40 tion and even create challenges for water purification processes.			The same trend (Figure 2) was mainly due to the stable removal ability of BAC and UF for organics, causing the removal restriction.			w14030367_perova	0
+Response to Reviewer 4 Comments Point 1: Authors should avoid abbreviations in the title.			Point 3: References should be according to the journal format.			w14030367_perova	0
+Please consider these at end of this sentence……The oxidation method exhibited a fast reaction speed and high removal efficiency…(i) Role of nanomaterials in the treatment of wastewater: A review (ii) Advances and challenges in developing efficient graphene oxide-based ZnO photocatalysts for dye photo-oxidation.			Response 14: Thank you very much for your kind reminder.			w14030367_perova	0
+If possible, please add at least one paragraph.			The study was carried out in the pilot-scale BAC-UF process with a treatment capacity of 0.16 m3/h.			w14030367_perova	0
+The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water.			Point 3: Line 46-52: & line 84-85 “Ultrafiltration (UF) as an emerging alternative technology to conventional water treatment processes, has been widely used to remove pollutants such as particles, colloids, bacteria, and viruses, thus reducing the risk of water-borne diseases [10].			w14030367_perova	0
+Water 2022, 13, x. https://doi.org/10.3390/xxxxx Academic Editor(s): Received: date Accepted: date Published: date Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations.			The authors reported the results with an average value and standard deviation.			w14030367_perova	0
+However, the long-term pilot-scale study is urge to be investigated.			Point 8: The main objective of the work must be written on the more clear and more concise way at the end of introduction section.			w14030367_perova	0
+Response 8: T Thanks very much for this comment.			The two-stage biofilms located in the activated carbon column and on the UF membrane synergistically, can be conducive to the removal performances.			w14030367_perova	0
+Response 10: Thank you very much for your kind reminder.			After the gas scrubbing and the hydraulic backwashing, the dissolved oxygen detection of the effluent was carried out, resulting in the above results for dissolved oxygen.			w14030367_perova	0
+Why is the standard deviation of Erythromycin showing a high value?			The comparison of previous research with existing research findings was added in the final part of the results and discussion.			w14030367_perova	0
+Response 7: Thank you very much for your kind reminder.			The BAC biofilter can remove the PPCPs, and then the following UF can reject micro-organisms and particles flowing out from the biofilter to ensure the quality of drinking water.			w14030367_perova	0
+Moreover, the authors present the results properly.			I am pleased to send you major level comments, there are some serious flaws which need to be corrected before publication.			w14030367_perova	0
+Point 14: Please use one style for units such as m3/h or m3h-1 Please revise your paper accordingly since some issue occurs on several spots in the paper.			Author Response Thank you very much for your support and help.			w14030367_perova	0
+Energy Conversion and Management, 235. Panagopoulos, A. (2021).			Results seem to be related to a unique experiment.			w14030367_perova	0
+These 38 PPCPs generally present in waters at trace concentrations, ranging from several ng/L to 39 thousands μg/L [4].			After the gas scrubbing and the hydraulic backwashing, the dissolved oxygen detection of the effluent was carried out, resulting in the above results for dissolved oxygen.			w14030367_perova	0
+Raw water quality parameters of river water (significant value, 0.05).			I can see very well-defined experiments and produced good results.			w14030367_perova	0
+“The combined process with ultrafiltration may be another promising choice, featuring a comparable removing performance as the nanofiltration and low operational cost”.			Environmental Science and Pollution Research, 1-14.			w14030367_perova	0
+So I had to acquire the Florida soil properties information by website.			Journal of Contaminant Hydrology,2015,177-178（June–July）:85-92.			w14081258_makarova	0
+Author Response Dear Editor： Thank you very much for the comments of my manuscript.			At first, the soil was 0.3 mm sieved, but the big particle size was not appropriate for the soil ultrasonic treatment of NP extraction.			w14081258_makarova	0
+Please feel free to contact me if any questions.			Consequently you excluded an important part of the active soil from your study, therefore your resuts are very limited.			w14081258_makarova	0
+I strongly recommend the authors to analysis the biomass of the soil samples R: Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential.			Indeed, the characteristics of the soil are too limited.			w14081258_makarova	0
+Yours sincerely, Shiyu Wang [email protected] Author Response File: Author Response.docx			Yours sincerely, Shiyu Wang [email protected] Author Response File: Author Response.docx			w14081258_makarova	0
+The main reason is that this experiment was conducted during my being in University of Florida, USA as a visiting scholar.			Please feel free to contact me if any questions.			w14081258_makarova	0
+The basic physi-chemical parameters of soils should be given.			The irrigation water physi-chemical parameters should be given.			w14081258_makarova	0
+Your careful review of this manuscript is highly appreciated.			(+86)13910796093 March 27 Dear Editor and Reviewers： Thank you very much for the comments of my manuscript.			w14081258_makarova	0
+(+86)13910796093 Apr 2 Dear Editor： Thank you very much for the comments of my manuscript.			The following are the details of the revision and marked red in the “resubmit” version.			w14081258_makarova	0
+I have carefully read the comments and made a revision.			I have carefully read the comments and made a revision.			w14081258_makarova	0
+Ecotoxicology and Environmental Safety 158 (2018) 28–36.			R: It has been given in Supporting Information 2.			w14081258_perova	0
+Yours sincerely, Shiyu Wang [email protected] Author Response File: Author Response.docx			Figure 1 legends of axis need font size larger, please make readable the figures R：Accepted and revised in L204.			w14081258_perova	0
+R: It has been given in Supporting Information 2.			Your careful review of this manuscript is highly appreciated.			w14081258_perova	0
+Actually, the soil samples were 0.25 mm sieved.			Figure 1 should be revised according to kinect model.			w14081258_perova	0
+I strongly recommend the authors to analysis the biomass of the soil samples R: Indeed, in this study, the analysis of biomass and microorganism of the soil samples is essential.			(+86)13910796093 March 27 Dear Editor and Reviewers： Thank you very much for the comments of my manuscript.			w14081258_perova	0
+However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.			Shiyu Wang, Wenyong Wu*, Fei Liu, Shiyang Yin, Zhe Bao, Honglu Liu.			w14081258_perova	0
+However, the information I can acquire from the website was limited, which were shown in Supporting Information 2.			The following are the details of the revision and marked red in the “resubmit” version.			w14081258_perova	0
+The following are the details of the revision and marked red in the “resubmit” version.			All the isomers were revised according to kinect model except some isomers, such as NP2 NP5 and NP11.			w14081258_perova	0
+Sorption and desorption behaviours of 4-nonylphenol on reclaimed water-irrigated soils.			Indeed, the characteristics of the soil are too limited.			w14081258_perova	0
+Then the soil was 0.25mm-sieved, which was suitable for the NP extraction.			Comments I do not understand why soil samples were 0.3 mm sieved.			w14081258_perova	0
+Indeed, the characteristics of the soil are too limited.			But for this study, the degradation was completed within 30 days in both of the two soils.			w14081258_perova	0