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Comment 35: l. 500 : The chemical shift range is more than from 1 to 5.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
l. 725 possibility of future
| 1 | 2 |
biom12060834_makarova
| 0 |
Comment 1: Unfortunately, its structure was not fully determined.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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:
| null | null |
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:
| null | null |
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
| null | null |
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?
| null | null |
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.
| null | null |
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.
| null | null |
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” ?
| null | null |
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:
| null | null |
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.
| null | null |
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.
| null | null |
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?
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
l. 228 GPC defined on l. 231
| 1 | 2 |
biom12060834_perova
| 0 |
Figure 6: Bacterial EPSs are normally composed of repeating units.
| null | null |
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:
| null | null |
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.
| null | null |
255, 286, and 441 et al.
| 1 | 2 |
biom12060834_perova
| 0 |
The studied EPS showed promising properties to use in the food industry.
| null | null |
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.
| null | null |
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.
| null | null |
Figure 3
| 1 | 2 |
biom12060834_perova
| 0 |
TOCSY and HMBC are mentioned in methods, but no results are presented.
| null | null |
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.
| null | null |
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:
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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:
| null | null |
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.)
| null | null |
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?
| null | null |
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:
| null | null |
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?
| null | null |
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.
| null | null |
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?
| null | null |
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?
| null | null |
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:
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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”?
| null | null |
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.
| null | null |
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.
| null | null |
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:
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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
| null | null |
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".
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
The supplementary figures are inaccessible.
| 1 | 2 |
brainsci12060789_makarova
| 0 |
There are also a number of sentences that are so complicated for easy read.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
The supplementary figures are inaccessible.
| 1 | 2 |
brainsci12060789_perova
| 0 |
There are some concerns need to be addressed.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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.
| null | null |
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 |
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