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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 |
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