Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: TEMPERATURE ALTERS THE SHAPE OF PREDATOR–PREY CYCLES THROUGH EFFECTS ON UNDERLYING MECHANISMS
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors reduced the number of free parameters in the model fitting from 8 to 6 by carefully fixing two parameters to reasonable values. They further added a test for model identifiability of this 6-parameter-model by using simulated data and checking if „true“ input parameters can be recovered by the fitting routine.

Yet, I as far as I understand from the Matlab code, the simulated data does not contain any stochasticity (e.g., observation error). If this is the case, it is no surprise that the model fitting will return the input parameters exactly (Fig. S1) and the fitted trajectory perfectly matches simulated data. In a strict sense, this does not indicate model identifiability, it merely shows that the optimizer correctly converges to the minimum of the deviance function. Model identifiability would require that, even including some noise, this minimum is still close to the true input parameters.

I suggest that a little observation error is added to the simulated data to test this. Maybe this does not require another round of reviews if the results don‘t change. But it would rule out any concerns about biased or redundant parameters and non-identifiability.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE LEVEL OF PUTATIVE CAROTENOID-BINDING PROTEINS DETERMINES THE BODY COLOR IN TWO SPECIES OF ENDEMIC LAKE BAIKAL AMPHIPODS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article structure follows conventional standards of scientific publication (good written English, figure quality, article structure etc.), but the authors include elements that should be provided in the Material and Methods section within the Results section, which is confusing.
Experimental design: The research question is not well defined and this evident for the introduction and the discussion section. The authors are not clear about what they are studying, which is the molecular formation of colour, but not the polymorphism. Their study objective to “uncover the mechanism underlying the formation of color polymorphism in E. cyaneus and E. vittatus” is incorrect, because they have investigated the molecular mechanisms of color formation (correctly stated at the beginning of the discussion). Consequently they authors are also drawing false or unsupported conclusions about the role of crustacyanins in colour polymorphism. The research gap is stated (lack of knowledge concerning colour formation in amphipods), but this is partly gone to water with the considerations about polymorphism. For parts of the study, the technical standards are perhaps suboptimal and need confirmation by additional work.
Validity of the findings: The novelty of the findings is limited as the mechanisms of colour formations are basically known and have been more or less confirmed here for amphipods. Some of the conclusions are, however, not valid as much as polymorphism is concerned. The authors have been comparing blue and orange morphs of two gammarid species. I do not think that this can be considered studying the mechanisms underlying polymorphism in these species and the focus should, therefore, rather be on the molecular role of crustacyanin in the formation of different colours in amphipods as compared to other crustaceans. With their focus on polymorphism the authors also tend to draw false and/or unsupported conclusions about the role of crustacyanins in producing highly polymorphic amphipods (for instance there are no data whether different populations of the studied amphipods would vary in polymorphism, as could be established by a Shannon-Wiener index, and how this relates to variability in the expression and concentrations of crustacyanins in the various morphs).
Additional comments: The manuscript PeerJ 44934-v0 describes the contribution of presumed amphipod crustacyanins to the formation of different colour morphs in two grammarids of Lake Baikal. The authors investigate the molecular mechanisms of colour formation in two species of the genus Eulimnogammarus from Lake Baikal, which has a particularly rich and polymorph diversity of amphipod species. The manuscript is well written, but the structure lacks clarity, particularly because the Material and Methods section lacks information that is contained in the results part. Furthermore, the objectives of the study are not clearly defined. Especially, the expression “mechanisms forming … diversity” or “mechanism underlying the formation of colour polymorphism” are, to my point of view, misleading. The scope of the study is much more limited and describes a putative mechanism of how different colours are formed in these animals, which, of course is a prerequisite for the observed colour polymorphism, but it is not the mechanism responsible for the polymorphism. Knowing how the colours are formed does neither explain within population variability in colour morphs, nor why these amphipods produce so many different colour morphs. In other words, it does not answer the question why the many different amphipod species in Lake Baikal are highly polymorphic. The mechanisms for this remarkable inter- and intraspecies polymorphism are not the capacity of forming different colours, but rather the interactions of these amphipods within a population and with their specific environment: location of habitat, depth, temperature, exposure to different predators, i.e., ecological and population parameters that trigger colour polymorphism.

Considering the article structure, the introduction describes this specific polymorphic composition of the vast amphipod fauna in Lake Baikal, which provides the background for the study. This done, however, too extensively and, more importantly, it should be clearly distinguished from the question of how the colours are formed. Within the introduction, the authors start with some information about the elements of colour formation (carotenoids, i.e., astaxanthin, crustacyanin) and their supposed roles in colour formation, but this aspect, which is fundamental to study, needs to be treated in more depth (e.g. see Cianci et al., 2002, Wade et al. 2005). The authors should inform how and where colours are formed in crustaceans. They should also point to the fact that they are dealing with morphological shell coloration. With regards to the method section, it would be interesting to know, what the authors are actually measuring (carotenoids from the shell, whole body, haemolymph) and how the haemolymph concentrations relate to the concentrations in the cuticle, which eventually determine crustacean shell colour. The authors, however, switch to the polymorphisms in Lake Baikal amphipods. What is further lacking in this context is a link how the authors consider the molecular mechanisms of colour formation to be linked to the polymorphisms. Hence I would recommend beginning the introduction with some remarks on the astonishing variability of coloration amongst Lake Baikal amphipods and the intraspecies polymorphism making it worthwhile to study the phenomena of colouration in these amphipods. The authors should explain to the reader what is the nature of this coloration (morphological pigmentation of the cuticle incorporating pigments from the epithelium) and how astaxanthines and crustacyanins interact with each other to produce different colours. Then they could continue with their statement, that a lack of knowledge exists with regards to these pigment and protein molecules in amphipods, providing the justification for the study.

The Material and methods section also lacks clarity. Unfortunately, a good amount of relevant information has been included into the results section. For instance, l. 196-198 explains the amount of animals samples, how they were classified and sorted, which is information that should be provided in ‘Animals and sampling’, as should be the information provided in l. 203-205. L. 206 talks of an experimental setup, but from the Material and methods section it is not clear that an experiment was carried out and what would be the nature of it. That animals were photographed after acclimation period also is methodological information. The paragraph ‘Carotenoid measurements’ mentions ‘samples’, but the authors do not specify what type of samples. From the results part it becomes clear that these samples are whole animals that were fixed in either ethanol or liquid N2. As the final colour is formed in the cuticle, why do the authors not study the carotenoids in the cuticle (extraction from shell as in Wade et al. 2005)? Similarly, why are crustacyanins from the haemolymph quantified although these are, most likely not involved in colour formation in the cuticle and why do the authors not extract crustacyanins from shell (e.g. Cianci et al., 2002). Can the authors ensure that both levels of crustacyanin are at least correlated? From the paragraph on electrophoretic methods it remains unclear what the 1D- and the 2D-PAGE are used for? It is quite surprising to learn from the results section that 2D gel electrophoresis is only used for a kind of pseudo-identification. Even if I follow the argumentation of the authors about the MW and pI matching theoretical values, it is difficult to accept that the authors have not confirmed the identity of the respective spots by MS/MS. The same holds true for the quantification of the 25kDa and 15kDa bands. Please understand that I do not question the assumptions made. The argumentation made by the authors is plausible (l. 249-250). Nevertheless, you are more or less blindly quantifying bands the nature of which has been inferred from secondary information. Furthermore, the manuscript would have gained impact if the authors could confirm the AA sequences and compare them with those retrieved in genbank. It should not be too difficult to find a group of mass spectrometrists that would analyse the respective spots. To my liking the proposed article presents too much ‘predicted’, suggested and ‘putative’ findings, which significantly lessens its overall impact. Given the coherence of the findings, I do not doubt the general conclusions of the study, but the relatively simple mass spectrometric analysis of four spots plus four bands would greatly increase the novelty, and thus the value of this study! Furthermore, the authors should demonstrate that their quantitative analyses from heamolymph do indeed reflect the levels of crustacyanins in the shell/cuticle, to ensure the validity of their measurements. Again the results are coherent and the conclusions drawn are plausible, but not verifying the correlation between the concentrations of crustacyanins in the cuticle and heamolymph and how these are related it is at least contestable. According to Ertl et al. (2013) the major site of crustacyanin expression is the endocuticle/outer epithelium and hence they are likely to be synthesised therein, or according to other sources in the subepidermal adipose tissue. But Ertl et al (2013) also found crustacyanin to be expressed in other tissues. It is therefore not straightforward to measure haemolymph crustacyanins as a surrogate of cuticle crustacyanis. It is also unclear how various physiological parameters or moult status would influence the concentrations of circulating crustacyanin. Hence, numerous open-ended questions may cast doubt over this method.

Eventually, the discussion is not strictly related to the role of astaxanthine and crustacyanin in the formation of shell colour. Here the manuscript suffers from the same lack of clarity already stated for the introduction and enters into a discussion about, how colour may change over lifetime, ecological aspects that contribute to polymorphism in Eulimnogammarus and relates the presence of crustcyanins to this polymorphism. To my point of view the authors mix the mechanism of forming blue colour in crustaceans, with mechanisms that produce the observed polymorphism. This is by no means the same! Furthermore, the authors claim that not the carotenoids are responsible for the colouration, but they are not precise about their comparison: the amount of asthaxanthin in the cuticle is certainly responsible for the degree of coloration between paler or more intensively coloured (orange) morphs (e.g. Wade et al. 2005). When it comes to blue coloured morphs, this requires molecular modifications of the astaxanthins to obtain a different absorption spectrum and in this case the crustacyanins are crucial for producing blue colour. Hence the difference between blue and orange morphs does not stem from the amount of carotenoids and this would indeed not be expected. It is known that he crustacyanins change the colour of astaxanthins and the results of the present study confirms this principle for the studied amphipods, thus increasing evidence that this is a general mechanism, even if some difference in the crustacyanin sequences exist. The authors should rather discuss these molecular mechanisms, which they have been studying. A good example is given by the penaeid shrimp F. merguiensis (Ertel et al. 2013). As the authors mention in their discussion the formation of colour morphs is a complex interplay between astaxanthin, crustacyanin and further factors. Hence, their conclusion that the carotenoid content is unimportant for the diversity of colour morphs (l. 312) cannot be generalised from the results of this study.

Furthermore, the capacity of forming blue, orange and other colours is not the reason for the observed polymorphism. The crustacyanins are simply a necessity to form complexes with the asthaxanthins that bend them into different orientations so as to change their absorption and produce different colours. But why these colours are produced in some morphs or in some body parts and not in others has not been studied and the discussion of these aspects is, therefore, not supported by any data of this study. The authors do not produce results that could explain the reasons for the polymorphism of these amphipods, i.e., why crustacyanins are produced at a higher degree in some morphs and to a lesser degree in others. Therefore, the discussion about polymorphism is distracting from the essence of the study, which was to confirm the role of crustcyanins in the formation of blue coloured amphipods. As the mechanism of blue colour formation has been described earlier (e.g. Krawczyk and Britton 2001, Chayen et al. 2003) and the extracted putative crustacyanins have been confirmed only indirectly the novelty of the proposed publication is, unfortunately, greatly reduced.

In summary, it is strongly recommended to repeat the gels with sufficient material for subsequent AA sequence analyses by MS/MS and to focus the introduction as well as the discussion on the molecular mechanisms of colour formation in crustaceans across different taxonomic groups (decapods, amphipods, …). If the MS/MS analyses cannot be carried out, the authors should at least whet the objectives of the study within the limits of what has been studied and avoid deviating from the molecular formation of different colours in the two gammarids into general considerations of polymorphism, which are not supported by the data. Nevertheless, the authors may, of course, highlight that the many highly polymorphic species of Lake Baikal provide a particularly interesting study subject.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE LEVEL OF PUTATIVE CAROTENOID-BINDING PROTEINS DETERMINES THE BODY COLOR IN TWO SPECIES OF ENDEMIC LAKE BAIKAL AMPHIPODS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The present study assessed the underlying molecular mechanisms that explain colour morphs in freshwater amphipods. The images are excellent and helpful, but the organisation of the Figures and the way in which the Results are presented is not so logical, for me at least.

1.	This entire Results section needs to be written more directly and in past tense. For example: L208-210 “We found that the individuals of this species had different colors, from completely blue to completely orange (Fig. 2A), and the metrics of R, G, and B colors of the pereon corroborated what was visible by eye (Fig. 2B).” and L221-225 “Six individuals from each end of the color distribution (as quantified using R/B values, Fig 3B) were used to quantify their carotenoid content. No significant difference in the overall carotenoid content of the body was found between these colour morphs (Fig. 3C), and thus indicated other mechanisms were involved.” There are many more examples throughout other sections, partly caused by the arrangement of data (see next comment).
2.	Once arriving at Fig 4 and 5, the Results begin to combine sequence comparisons and protein blots from one species, then repeats several colour analyses, carotenoid and protein extractions from another. In my opinion, it would be better to put the animal descriptions (for both species) up front in Figure 2, then organise the proceeding Figures more consistently to contain shared data from both species in successive Figures. This might be something like: PCA of RGB values in one Figure, carotenoid quantification in another, then protein blots. I question the need for phylogenetic analysis as part of these Figures, as this is covered in Figure 6. As presented, the organisation of the Figures does not allow for a very easy description of Results and leads to confusion in the Results section, plus considerable Discussion points in the Results. This suggested re-organisation is supported by the authors own statement on L277-279, that Results are similar for both species. Perhaps consider a combined Results and Discussion section?
Experimental design: An impressive series of experiments was conducted on a species with a distinct colour morph, with the clear goal to understand the role or protein bound carotenoids to produce their colouration. In addition, further investigation of CRCN sequences from a range of crustaceans was performed, and further resolved the diversity and evolutionary significance of this protein family. The authors should be commended for including their coding as part of the manuscript, this is often overlooked. The extraction of hemolymph from such small animals also shows great technical skill.

3.	Please provide a more detailed description of the method used to quantify colour from digital images.
4.	The methods are well explained but would benefit from clearly stating how many individuals were used for each method. This would clearly describe how many individuals were used for each of the colour quantification, carotenoid extraction or protein gel sections. If the same individuals were used in different sections this should also be made clear, as this can strengthen the associations between the data sections.
5.	Please describe the use of coloured dots in Figure 2 as part of the Figure legend. However, only two pale blue and intermediate blue/orange individuals were shown on Figure 2. I question the need, ability or importance to isolate them from the blue and orange forms. Were these individuals only defined by eye, or was there supporting RGB values that could classify them? Without support for the additional colours based on RGB quantification, it might be best to remove the two individuals from the other graphs and just use the individuals from the blue and orange dots.
6.	The field frequency data is useful, but less so the survival information provided as part of Supplementary File S3. If not replicated and significantly different between the two morphs (which was not stated or evident from the data presented), this would be the weakest part of the study and might be best left out so remove lines 203-206.
7.	Although quite distinct species, the past study by Mojib et al 2014 (Molecular Ecology 23: 2740-2756) should be included in the phylogenetic analyses and Discussion, and mentioned in the Introduction as well. The present work builds on this nicely to demonstrate the expansion of CRCN genes among animals.
8.	It was not clear whether the protein alignments used full length sequences, which could be made clearer either by denoting the corresponding residues from the reference sequence (as part of the Methods) or include the alignment file used for the phylogeny as a Supplementary file.
Validity of the findings: It is very interesting to me the appearance of several CRCN genes in diverse crustacean lineages, broadening the functional diversity of this family. The conclusion that these proteins are acting in the same way to produce colour despite considerable sequence diversity is solid.

9.	L43-48 – several studies in decapod crustaceans have shown that pigment levels are not correlated with body colour, particularly in response to environmental cues. The study by Ertl was the exception to others such as Tume et al 2009 (Aquaculture 296: 129-135) and Wade et al 2015 (Aquaculture 449: 78-86). Indeed, the latter study quantified CRCN abundance using an antibody
Additional comments: It would be interesting to know whether the carotenoids extracted from red morphs were esterified, as this was found to be the case for Penaeid prawns that had removed their crustacyanin. Perhaps more a comment for future work than a requirement for modifying this work, this can be done relatively simply using thin layer chromatography or HPLC if you have one.

Would be nice to crystalise these proteins, or maybe even just predict their 3D structure based on decapod CRCN!

Minor comments
•	Line 123 One blue and one orange individual was…..
•	Line 207 I would say this method was quantitative. You should remove “rough”
•	Exact significance values are not important, and it is sufficient to use a P value cutoff of 0.05 or 0.001. Non significant P values do not need to be reported at all
•	The arrows in Figure 4B don’t quite align with the protein spots
•	It would be helpful to define aLRT in the Figure 6 legend, and explain the associated red dots on the phylogeny.
•	Lines 360-364 has some awkward wording, perhaps divide into several sentences.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE LEVEL OF PUTATIVE CAROTENOID-BINDING PROTEINS DETERMINES THE BODY COLOR IN TWO SPECIES OF ENDEMIC LAKE BAIKAL AMPHIPODS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors write a correct scientific English that describes the research subject, the methods used as well as the results and corresponding discussion in a clear and unambiguous manner

The introduction of the revised manuscript is focused, places the research work in a clearly defined context and delivers the necessary background in a concise, but comprehensive way. The cited literature is pertinent and references the current state of the art.

The structure of the manuscript conforms the discipline norm of scientific publications in the field of life sciences and has been greatly for improved for clarity as compared to the original manuscript.

All figures are relevant and of exceptional quality. The figures are extensively labelled and described correctly.

The data supplied are ample and exhaustive. Extensive supplementary data are provided, but mass spectrometry data seem not to be included.
Experimental design: The manuscript describes original primary research that is within the scope of the journal.

The research question is now defined more clearly. The authors have clearly outlined the lack of knowledge for the taxonomic group subject to their investigations, as compared to the decapods, where coloration has been studied in more detail. Hence, the research question is relevant and meaningful. The results obtained in this study also underscore the research need with regard to the mechanism of colouration in species less well studied.

The investigations performed are scientifically sound and of high technical standard.

The methods are described with sufficient detail and should be reproducible. The material and methods section has been improved as compared to the original manuscript. Given the impressive amount of work accomplished at different levels (sampling, image analyses, protein biochemistry, molecular biology) this section naturally has to be quite voluminous.
Validity of the findings: The data supporting the conclusions have been provided exhaustively; they appear to be coherent and reliable.

The impact and novelty of the data is highlighted in the results and discussion sections. By including the results of mass spectrometric analyses, the manuscript has considerably gained in impact and novelty, because the results revealed an unexpected nature of the proteins. These surprising results open new research questions and should be very interesting to the scientific community working in the field. It is understood, that the new proteins revealed by this study, need to be confirmed and their role remains, for the time being, speculative. The authors have made this perfectly clear.

The conclusions in the revised version of the manuscript are well stated, clearly linked to original research question described in the introduction and based on the outcome of the study.
Additional comments: First of all I would like to express my respect for the impressive amount of work the authors have invested into this study and congratulate them for the additional work that they have accomplished in a little while. This is a magnificent example of how to revise a manuscript! . By my reckoning, the investment into mass spectrometry was worth the while and has both increased the novelty and impact that the paper will have.
Indeed, the manuscript has exquisitely evolved and matured to an impactful research work that should make an excellent contribution to the scientific literature on colouration in crustaceans. The authors have responded perfectly to the questions of the reviewers. For my part, I have no further questions or comments. I can only applaud the authors for the work they accomplished and wholeheartedly recommend the manuscript for publication!
However, I would suggest to improve the title and suggest something like “Putative carotenoid-binding proteins determine colour formation in two species of endemic Lake Baikal amphipods”. Furthermore, there is a typo in the abstract (L.41 “then” should be “than”). I would also like to suggest some corrections to the figure legends:
Rather use the term “Representative photographs” in the legend to figure 4, as in the legends to figure 1, 2 and 3 and replace “example” by "representative", i.e., "representative 2D-PAGE/1D-PAGE". Perhaps specify (MS/MS) for the mass spectrometric analysis. In the legend to figure 5, replace “some amphipod species” by “selected amphipod species”.
I will take great pleasure to see this manuscript published and congratulate the authors for their work! The acknowledgements for reviewing this article are flattering, but would have not been necessary. I did my best and if this was helpful the sense of reviewing was fulfilled.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ALVEOLAR SOFT-PART SARCOMA (ASPS) RESEMBLES A MESENCHYMAL STROMAL PROGENITOR: EVIDENCE FROM META-ANALYSIS OF TRANSCRIPTOMIC DATA
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The author reports here that ASPS resembles a mesenchymal stromal progenitor at the mRNA level. The study is well illustrated and documented, and the conclusions are certainly supported by the data presented. However, some parts of the manuscript need revision, before it can be accepted for publication.

#1 Materials and methods are too short and not sufficient to understand.
In Table 1, the author has listed the top 50 transcripts overexpressed in the ASPS-1 cell line relative to the average. Is it an appropriate analysis method to use the average value of completely different samples? A bias exists in the number of cell lines. The number of Ewing sarcoma cell lines is 22; on the other hand, the number of DDLS cell lines is only 2. The average value reflects the majority of data (Ewing sarcoma gene expression value). The author has to better compare with ASPS-1 individually for each cell line type.

#2 In Figure 1A, cell lines from the same tumor are grouped into separate clusters. For example, fibrosarcoma and osteosarcoma cell lines are classified into various clusters. The author should check the bootstrap value to confirm the reliability of the branches of the phylogenetic tree. The author may also attempt nonhierarchical clustering, such as PCA.

#3 In Figure 3, the letters are too small and the graph is complicated. The unit of the vertical axis of the graph is not listed.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ALVEOLAR SOFT-PART SARCOMA (ASPS) RESEMBLES A MESENCHYMAL STROMAL PROGENITOR: EVIDENCE FROM META-ANALYSIS OF TRANSCRIPTOMIC DATA
Review round: 1
Reviewer: 2

Output:
Basic reporting: This study investigated the many genes expression levels in ASPS compared to other sarcoma cells, normal tissues etc. They showed ASPS transcriptome resembles that of mesenchymal stromal progenitor. Therefore, their study based on the data from gene expressions, I thought that it had a certain level of reliability.
Experimental design: Experimental design had no problem. I thought they should do the experiments such as RNAi for comparison between the gene expressions of ASPL-TFE3 knockdown and not. But they commented about the point in the Conclusions. ASPS sarcoma cells have chromosomal translocation of ASPL-TFE3, so, in the cells, various changes could be occurred. So, it is very interesting that they investigate the changes of gene expressions based of RNAi experiments (ASPL-TFE3 knockdown).
Validity of the findings: Their findings had a certain level of reliability.

There are several points that need minor changes as followings;
・In line 53, ASPSCR should be changed to ASPSCR1.
・In line 309, ASPL-TFE should be change to ASPL-TFE3.
・Author should use the unified abbreviation such as MSC in line293-302.
・In Figure2, Table1, 2, author should correct the spelling of “fifty”.
Additional comments: I thought this study is acceptable for this journal. ASPS sarcoma cells have chromosomal translocation of ASPL-TFE3, so, in the cells, various changes could be occurred. So, it is very interesting that they investigate the changes of gene expressions based of RNAi experiments (ASPL-TFE3 knockdown).


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ALVEOLAR SOFT-PART SARCOMA (ASPS) RESEMBLES A MESENCHYMAL STROMAL PROGENITOR: EVIDENCE FROM META-ANALYSIS OF TRANSCRIPTOMIC DATA
Review round: 2
Reviewer: 1

Output:
Basic reporting: I have no comment.
Experimental design: I have no comment.
Validity of the findings: I have no comment.
Additional comments: Authors change the points of reviewers, and this article is acceptable for this journal.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE COMMON PERSONAL BEHAVIOR AND PREVENTIVE MEASURES AMONG 42 UNINFECTED TRAVELERS FROM THE HUBEI PROVINCE, CHINA DURING COVID-19 OUTBREAK: A CROSS-SECTIONAL SURVEY IN MACAO SAR, CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This manuscript is addressing an interesting question: to identify common characteristics of personal behavior and preventive measures of 42 uninfected travelers from the Hubei province, China during COVID-19 outbreak. Further, the authors perform a comparison of personal preventive measures before and during disease outbreak,  showing increased alert and practice of personal protection and hygiene during the spread of SARS CoV 2. The result will contribute to the improvement of epidemic prevention mesures policies in future and individual health guidelines.

Minor comments:

1. Were IgM and IgG titers quantified on travellers? If yes, please add the information to the text.

2. line 76. "After arrival to Macao, all 57 citizens were sent to public health clinical center for a 14-day quarantine. A total of three serial nasopharyngeal swabs were obtained on day 2, day 7 and day 13 for viral RNA detection by real-time RT-PCR techniques, which were all negative (100%). Sera antibodies of SAR-CoV-2 were tested with all negative results (100%) on day 14 before citizens released from quarantine (6)." This reference do not mention serological tests as indicated on text.

3. All tables needs to be revised. To reduce the excessive unnecessary informations on tables, remove % symbol from first column of all tables and number 42 from all results of table 1 (it is redundant once the number is at the topic Proportion (n=42).

4. Table titles should be presented once. The title is rewrited inside the tables.

5. Table 1. Once sex/gender may influence on adoption of prevention measure and epidemiological data,  please organize the second column in male and female

6. Table 1. "Presence of chronic disease(s)" put the none % at the result

7. Table 1. turns "current smoker" a subtopic of "Tabagism" as the "ex smoker" and "never"

8. Table 2. Remove "No" column and reorganize "yes" data on male and female

9. Table 3. set the * as statistics significance at the table footer. It is not clear the P value numbers.

10. line 97 "A total of 42 effective questionnaires were analyzed in final after exclusion of 14 infants and
98 children with age less than 15 years old and missing of one questionnaire (response rate: 97.7%)". It is not clear why some topics at table 3 has only 40 or 41 instead 42 total N?

11. Some numbers of table results are not discussed. The discussion is attentive mainly to lockdown measures and the paper has demografic data (table 1) such as tabagism, chronic diseases, education that may reflet the ausence of COVID19 positive among these group.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE COMMON PERSONAL BEHAVIOR AND PREVENTIVE MEASURES AMONG 42 UNINFECTED TRAVELERS FROM THE HUBEI PROVINCE, CHINA DURING COVID-19 OUTBREAK: A CROSS-SECTIONAL SURVEY IN MACAO SAR, CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article meets the standarts.
Experimental design: The article meets the standarts.
Validity of the findings: I believe that in the "conclusion" the last sentence (line 201) will look better if it is written clearer, more understandable and highlighter.
Thanks.
Additional comments: Thank you


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE COMMON PERSONAL BEHAVIOR AND PREVENTIVE MEASURES AMONG 42 UNINFECTED TRAVELERS FROM THE HUBEI PROVINCE, CHINA DURING COVID-19 OUTBREAK: A CROSS-SECTIONAL SURVEY IN MACAO SAR, CHINA
Review round: 1
Reviewer: 3

Output:
Basic reporting: There are several areas where the authors need to improve writing.  For instance, line-81, line-176, line-186, etc.
Experimental design: More details are required for the survey design and its implementation.
Validity of the findings: In general, the findings seem to correspond to intuition.  However, in Table 3, why "clean and disinfect house regularly" drops from 36 to 31 for the before and after cases.  Is there any specific reason behind this?
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE COMMON PERSONAL BEHAVIOR AND PREVENTIVE MEASURES AMONG 42 UNINFECTED TRAVELERS FROM THE HUBEI PROVINCE, CHINA DURING COVID-19 OUTBREAK: A CROSS-SECTIONAL SURVEY IN MACAO SAR, CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: NO COMMENTS
Validity of the findings: No comments
Additional comments: The authors clarified all questions and made the suggested changes to improve the text quality.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: THE COMMON PERSONAL BEHAVIOR AND PREVENTIVE MEASURES AMONG 42 UNINFECTED TRAVELERS FROM THE HUBEI PROVINCE, CHINA DURING COVID-19 OUTBREAK: A CROSS-SECTIONAL SURVEY IN MACAO SAR, CHINA
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Perform another round of proof-reading and grammatical check.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HOW CAGED SALMON RESPOND TO WAVES DEPENDS ON TIME OF DAY AND CURRENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: Is it a limitation to one of the two methods applied for the evaluation of the objectives? Otherwise, please explain that acquired data were sufficient to extract the related results. At least, define entire time period that video recordings were acquired.
Validity of the findings: No comment
Additional comments: The aim of the study looks very interesting and provide state of the art knowledge on a very promising attempt to expand aquaculture towards a more sustainable way in the future.

In my opinion, manuscript should be accepted for publication, under some minor revisions.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HOW CAGED SALMON RESPOND TO WAVES DEPENDS ON TIME OF DAY AND CURRENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Basic reporting

This manuscript addresses an important issue in salmon aquaculture, and presents interesting results based on a valid methodology, so has the potential for publication. However, the work needs be presented in a much clearer fashion. It requires a lot of polishing and more attention to detail.

1)	The Discussion section could be a lot more focused.

For instance, a discussion could lead with the main points shown by the work (e.g. https://www.biosciencewriters.com/How-to-Write-a-Strong-Discussion-in-Scientific-Manuscripts.aspx), and then elaborate on these within the context of the literature (including a section on potential weaknesses/limitations). The discussion in this manuscript is not well organized. For instance, the sub-sections are “Daytime behaviour", “Night time behaviour" and “Video data” – the first two sections are on behavior, so surely the final section (“Video data”) should also be named in the same way? – for instance “Swimming behavior”

The authors open with a line (“When there is enough light so that salmon are able to easily see each other and their surroundings, they need less space to avoid collision.”) that feels like it could be part of the Introduction section. It is not a strong opening. What about a paragraph summarizing the key findings of the research, followed by an expansion on these in subsequent sub-sections?

The authors often use the present tense to describe their results in the Discussion section, which implies that that they are talking in general, when they are sometimes clearly referring to their own work. For example, L409 “In strong currents, fish mostly move downwards as opposed to during the day with the exception of fish at the front of the cage in short period waves.” Or L460 “There is a need to seek deeper water where waves are weaker, but there is also a need to move away from the net, which again may be encroaching on the usual space if there is any net deformation.” I strongly suggest that they change to pass tense except when the authors talking in general.

A lot of the Discussion reads a bit like the Results section. For instance, a lot of the text on daytime and night time behavior is just repeating what was in the Results section.

The Discussion is also sometimes a bit repetitive. For instance, the final paragraph on video data begins with “This indicates a preference for avoiding the net and orienting according to wave rather than current when waves are large.” (i.e. begins with net avoidance in large waves) and ends with “This makes sense considering the increased risk of collision with the net that is likely when waves are large and unpredictable.” (i.e. ends with net avoidance and large waves). It could be much more succinct.

The Discussion is also quite difficult to read, with a lot of description of results that could be summarized better. The authors could consider introducing each paragraph in the Discussion with a topic sentence, which summarizes what the paragraph is about. For instance, what about merging the “Daytime behaviours” and “Night time behaviours” sub-sections, and introducing the text with a clear statement (“Fish were more spread out during night than day”), and then expanding upon this? Framing with an introductory topic sentence makes it easier for the reader.

2)	There are too many figures.

In particular, figures dealing with shoal position and distribution (Figure 8, 9, 10 and 11) are difficult to compare with one another because they are separated across multiple pages. Could these be integrated into one figure (each figure would be a panel) as follows?
i.	Upper left panel – front of cage, daytime
ii.	Lower left panel – back of cage, nighttime
iii.	Upper right panel – front of cage, daytime
iv.	Lower right panel – back of cage, nighttime
This would allow the reader to directly compare between the front and the back of the cage, and between daytime and nighttime

Figures 1, 4, 5 and 6 could potentially be in the supplementary material.

3)	The English is generally ok, but could be a bit more formal and precise.

The authors should use past tense when describing their results (and when referring to them in the Discussion). Sentences could be made more direct, with fewer clauses. I go into more detail in General Comments below.
Experimental design: I can find no big problems with the experimental design. However, the authors could be a bit clearer about the statistical methods.

1)	How did they find the upper and lower bound to be used in the linear models?

2)	The authors need to also include some text of how they qualitatively chose wave size for analysis of the video data (first addressed in the Discussion on L449)
Validity of the findings: The findings are valid
Additional comments: 1)	The abstract needs improvement.

There are seven introductory sentences before the authors get to their study, which is too long.

Also "This study employs two major fish monitoring methods to determine the ability of Atlantic Salmon (Salmo salar) to cope with wavy conditions”. Add “…in offshore fish farms” (otherwise it is too general).

“Video cameras are used to monitor … proximity to camera locations” Would “spatial distribution” be better than “proximity to cameras” (the study is not interested in proximity to camera locations per se).

“The results indicate interacting effects between wave parameters and currents.”. So one would expect the following sentence to lead on from this (i.e. on interaction between waves and currents), but it then talks about fish behavior instead.

“In weak currents, fish generally move further down in taller waves, but stronger currents generally caused fish to move upwards.” This has too many clauses and is difficult to read. Change to “Fish generally moved downwards under conditions of weak currents and taller waves, …”?

The abstract does not mention day night effects, which is a key part of the findings.

2)	The authors should make sure they are presenting scientific units correctly. E.g. write  “14-20 s”  rather than “14-20sec”, “30 cm s-1” rather than “30cm/s”

3)	Regarding sub-headings in Material and Methods, could the opening paragraph and the sub-sections “Waves” and “Currents” all merged into the same subsection? e.g. “Study Area” (because they are all describing the study area).

4)	I suggest changing the order of the figure panels so that they are consistent with the order of the text (L338-350). e.g. they are referred to in the text in the following order: Fig 13 D, Fig 13 A, Fig 13 C, Fig 13 B, whereas it should be A, B, C and then D. Also, the text on L340 implies that Figure 13 only refer to then fish were shoaling (“but when fish did shoal”)? If so this should be mentioned in the figure caption.

5)	In the Materials and Method section, I suggest moving the video camera text (L179-186 and L224-239)  to be after the echo sounder text, so that it is in the same order to that where it is presented in the results.

6)	The figures could be improved.

Font sizes need to be increased.
Numbers should not be engineering notation. Axes should be capitalized (e.g. “Mar 04”, “count”).
Figure 1: the axes need labels, and the caption should specify “wave period (Tp)”. Tp (not T) should be used in the figures for the sake of consistency.
Figure 3: Can a scale be included? Or could this be superimposed onto Figure 1.
Legend for plots of shoal position and distribution: Suggest in full “Wave height (Hs)” “Wave period (Tp”).

7)	Minor points

There are quite a few small changes that could improve clarity, including:
L45: “44 %”?
L69: remove comma after “there is a risk”
L93: should be “a recent study has found”
L122: specify the country
L124 better as “The maximum wave height measured at the most exposed cages is 5.3 m”.
L127: remove “the one”
L155: remove italics from “coming”
L175: “these did not”, not “these do not”
L225: “at a two hour interval”, not «with…»?
L321: “were distributed nearer to the surface” or “were distributed higher within the water column” better than “were generally higher up”
L334: Could be better expressed: “Video data indicated a reduction in tail bear frequency with increasing wave size”
L359 “muddies the waters” is too informal. “makes interpreting the data more difficult”?
L369 “cast more light” is too informal. “provide more information”?
L397: “However, the number of fish in the shoal does not explain the depth change as all of the shoal at the back is above 8m depth in large, long period waves and strong currents.” Not very clear and needs rephrasing.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HOW CAGED SALMON RESPOND TO WAVES DEPENDS ON TIME OF DAY AND CURRENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Nothing to add
Experimental design: Nothing to add
Validity of the findings: Nothing to add
Additional comments: Nothing to add


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HOW CAGED SALMON RESPOND TO WAVES DEPENDS ON TIME OF DAY AND CURRENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The reporting is now clear. The results and discussion have been improved. Some minor improvements to some figures could be made (see General comments for the author)
Experimental design: The experimental design is ok. The authors acknowledge potential limitations (mistiming between ADDP and video data, and their lack of information on cage deformation). Other aspects are fine, and are discussed well.
Validity of the findings: The findings are valid.
Additional comments: This manuscript has been greatly improved. The authors have done a good job in presenting the results, and the discussion is now a lot clearer and better structured. It presents interesting and useful findings, so I can recommend this for publication.

However, I can’t find a list of captions for the supplementary figures. Were these lost on the upload? I think these will have to be checked, so this part of the manuscript might require minor changes that probably do not require re-review.

There are a few (mainly optional) minor points that could be addressed, and some figures could be improved.

Minor points

L145: “(here meaning the directions between east and south)”. Probably not necessary. Remove?

L304: “Data are presented separately for T1 and T2 (“front” and “back”) echo sounders.” I don’t think this is necessary. Remove?

The authors should check that there is always a space between value and unit. e.g. “10 m”, not “10m”.

A very minor point: the final sentence of the conclusion uses “smolt”. Is this appropriate? Is “post-smolt” a better term for a salmon in a fjord, with “smolt” referring to the fish in freshwater?

Figures

Fig 2: the caption could include “PRC Nautical Area Scattering Coefficient (PRC_NASC)”.

Fig 5B should be same size as the other panels.

Fig S2 should have “Temperature (oC)” under upper panel. The x-axis values should ideally have the same precision (e.g. not 7.60 and 7.7 together).


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1) English needs to be edited by a native speaker. There  are many sentences that are not clearly written or ambiguous.
2) References: There are numerous mistakes in style and content such as author names, year of publication, and so on. In this connection, there are mistakes in citing references in the text, e.g., line 194.
3) Introduction: Please explain why rabbit melanocytes need to be studied regarding the role of KIT when many studied have been done on its roles in mouse and human melanocytes. Any advantages or any specific purposes? Also, what the KIT is needs to be briefly explained along with citation of proper reviews or articles regarding this point, such as a brief summary of signaling pathway in melanogenesis.
4) Discussion is not up to the standard of Peer J. There are many sentences whose meanings are not clear.
5) In Discussion, please discuss how KIT acts on proliferation and melanogenesis in rabbit melanocytes in comparison with mouse and human melanocytes. Are they similar or different?
Experimental design: 1) Materials and Methods section is written adequately.
2) Experimental design is acceptable. However, Results section needs to be written more in detail.
Validity of the findings: The authors discussed melanogenesis in term of eumelanin to pheomelanin ratio. However, from the results of this study, discussion in this regard is not fruitful. But speculation is welcome (to be identified as 'speculation').
Additional comments: Two additional minor comments:
1) Some names of Rex rabbits are strange: protein chinchilla and protein yellow. Are they well-known? Any references to color of those Rex rabbits?
2) I believe that scientific names of species (scubas Oryctolagus cuncils) are written in italic.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 1
Reviewer: 2

Output:
Basic reporting: Hu et al. revealed that KIT could affect the expression of pigmentation-related genes and affect the proliferation and apoptosis of melanocyte. The results are not enough to support their conclusions. The original data are needed. The use of English should be improved.
Experimental design: 1. Fig 2. The author only detected the expression of the genes at the transcriptional level. They should also detect the expression at the protein level with WB.
2. Fig3. The author detected the proliferation with CCK-8 and detected the apoptosis with FACS. Only one preliminary screening method does not provide enough proof for the conclusion. The expression of proliferation and apoptosis markers are needed.
3. The authors revealed a phenomenon without any potential mechanism. To make the story complete, they could detect the expression of some potential molecules at the same time, and speculate on the potential mechanism.
Validity of the findings: 1. For nearly all the statistical figures, the author only provided the mean and SD. They should provide the original data.
2. Fig 1a. Please provide the uncropped figures
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors consider the role of KIT in pigmentation of rabbits.They chose to focus on KIT as a pivotal gene of melanin synthesis, modulated its expression in cultured melanocytes and observed the melanogenesis related genes expression after KIT modulation.

The introduction would benefit from more details and references around melanin synthesis and the genes involved. For example, it would be interesting to detail why MITF, TYR, PMEL are studied here, and how their expression is regulated by KIT. The whole melanogenesis being regulated by KIT and MITF, among other molecules, the authors should highlight the connections and regulations of these genes. In this regard, nearly half of the discussion should be moved to the introduction part.

In the discussion (line 220), the authors can not conclude that "KIT directly affected melanin-related genes at the transcription level". The showed a correlation of expression between those genes, but not a direct effect of KIT modulation.

ENglish should be revised, a few typos and grammar issues can be found in the manuscript.
Experimental design: The experimental design is relevant regarding the question defined. However, more experimental details should be provided by the authors.

Concerning the animals, pictures of each coat color should be provided.

Transfection experiments : How did the authors control the transfection efficiency?
how is the expression level of the genes (fig 2 and 3) corrected for transfection efficiency?
How was the transfection performed? in which culture medium?

Quantitative PCR : how did the authors check the quality of RNAs? How much RNA was used for cDNA synthesis? and how much cDNA for RT-PCR? Please provide the primer efficiency and concentration used.

Melanocyte proliferation and apoptosis : were the experimenst performed on several replicates of the transfection?

More generally, for each experiment, the number of samples used and number of replicates (technical, biological) should be precised,  as well as the statistics used.
Validity of the findings: The authors should explain the difference and added-value of the present study compared to their precious study (Hu et al., Biochemical Genetics, October 2019, Volume 57, Issue 5, pp 734–744), since this article deals with melanogenesis gene expression in rabbits with different coat colors. Were the seame samples/data used in the two studies?

The authors chose to clone the KIT gene in rabbit. The sequence they described was deposited in Genbank in 2017. SInce then, a detailed and annotated assembly was released (and accessible for example through Ensembl), and should be used by the authors to be compared to their sequence. What does the difference in size account for? Are there variation in nucleotide sequence or protein sequence compared to the reference assembly?

The gene expression of KIT and melanogenesis related genes was investigated in skin. these genes are specific to melanocytes, which are a minor population of skin. Also, in many species, coat color differences are due to gene expression differences and/or differences in amount of melanocytes in the skin. The authors should therefore provide a picture and estimation of quantity of melanocytes in the skins investigated. Without this data, teh authors can not conclude that KIT is less expressed, since the lower level could be due to a lower number of cells where KIT is expressed.
Isn't it possible to quantify the protein signal with the Wes?

Figure 2 : how was annotation of KIT performed?

The authors should comment on the transitory effect of the transfection. They analysed expression 48 or 72 hours after transfection, so how did they ensure i) that the transfection was efficient and ii) that cells were still transfected after 48 or 72 h?
Fig 2c : which siRNA was chosen?
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors addressed most of the concerns. There are still some concerns remained.
Experimental design: 1. The authors claimed that they performed WB with antibodies target TYR, MITF, PMEL, and DCT, but no result was obtained. Why? Can they submit the "negative result" as supplementary or just as raw data?
2. The authors did not respond to one main concern: " The authors revealed a phenomenon without any potential mechanism. To make the story complete, they could detect the expression of some potential molecules at the same time, and speculate on the potential mechanism. "
3. Fig 4a and 4b, the melanin content can be seen directly in the concentrated cells ( Pls refer to DOI: 10.1016/j.jdermsci.2016.04.005), which will make the claim more reliable. Can the author provide the pictures?
Validity of the findings: Some legends are wrong or confused.
1. All the relative expressions were not illustrated clearly. Were they standardized by internal control?
2. The author should quantitatively analyze all of the WB results because the expression of internal control seems apparently different.
3. Fig 3d looks like WB results, but the authors claimed it to be a qPCR results. Fig 3e is apparently not obtained from the KIT-overexpressed group.
4. Fig 4e-4h was wrongly described in the legends.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors improved the clarity of the manuscript and structure regarding introduction/discussion.
Experimental design: Many comments were addressed to the authors regarding experimental set-up and results.

Some of the comments were not clearly answered (for example: primer efficiency for the qRT-PCR experiment, efficiency of the transfection, etc...) and must be stated to ensure validity of the findings.

Most comments got, however, a detailed response. Unfortunately, the authors did not include elements of their response in the corrected manuscript. The objective of the reviewers questions is to clarify the article and add essential technical elements to the article. Thus the authors should carrefully add technical details that were asked by the reviewers. For example it is crucial to add the number of animals studied per experiment, in the figure legend, and not only in supplementary data. Also, the differences between the KIT sequence obtained by the authors and the reference sequence should be described and discussed.

The supplementary data are quite complete, although it is difficult to follow; It seems that the numbers attributed to the excel sheets do not correspond to the figures numbers. And there are some missing data (for example, Wes quantification on fig3d)

Finally the authors should review the protocols they described in the Materials and Methods: for example, it is very surprising thet qRT-PCR experiments were performed with 10µM solution of  primers. I guess it is the initial concentration of primers. The information that is needed is the final concentration of primers in the PCR reaction.
Similarly, the quantity of siRNAs used or silencing should be given instead of a volume.
Validity of the findings: The results obtained seem relevant and sound. However, if teh authors do not provide the elements asked, the study can not be considered fully validated.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors addressed most of the concerns. There are still some concerns remained.
Experimental design: no comment
Validity of the findings: 1. It seems that the raw picture of figure 2f-GAPDH and the figure they used in the manuscript is not the same.
2. What do you want to express with Table 4? It seems that there was no data.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: KIT IS INVOLVED IN MELANOCYTE PROLIFERATION, APOPTOSIS AND MELANOGENESIS IN THE REX RABBIT
Review round: 3
Reviewer: 2

Output:
Basic reporting: Despite typos left, the article is now clear and quite well structured
Experimental design: Most questions regarding experimental design and methods where addressed
Validity of the findings: Fine
Additional comments: The authors addressed the majority of remarks. Their corrections improved the mansucript a lot.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HABITUAL AND READY POSITIONS IN FEMALE TABLE TENNIS PLAYERS AND THEIR RELATION TO THE PREVALENCE OF BACK PAIN
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article is written in English and  use clear, unambiguous, technically correct text. The article is conform to professional standards of courtesy and expression.
The article include sufficient introduction and background. Literature is appropriately referenced.

The structure of the article is in  acceptable format

Figures are relevant to the content of the article and appropriately described and labeled.

All appropriate raw data have are  made in accordance with of Data Sharing policy.

The manuscript include all results relevant to the hypothesis.
Experimental design: The manuscript is original primary research within Aims and Scope of the journal.
The manuscript should clearly define the research question. The knowledge gap being investigated should are identified.
The investigation has high technical standard.
The research were conducted with the prevailing ethical standards in the field.
Research procedures described very clearly.
Validity of the findings: The presented results were based on objective research and bring practical solutions to sports training of table tenis.
The data and conclusions are based on available in an acceptable discipline-specific repository. The data  are robust, statistically sound, and controlled.
The conclusions presented in the article contain the results, the following conclusions should be described in more detail from the three lines of text 338,339,340. The rest of the text from this section should be moved to the results and discussions section.
Additional comments: The manuscript presented is original contains, objective research, results are original and conclusions concurrently Apply the development of better training methods for famale playing table tenis.
However, it is necessary to rebuild the chapter of conclusions.
Results are not conclusions, so results should be removed from this chapter and the focus should be on the use of the results in practice.
The importance of the results obtained should be thoroughly discussed in terms of improving the efficiency of table tennis players' training and reducing pain felt during training.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HABITUAL AND READY POSITIONS IN FEMALE TABLE TENNIS PLAYERS AND THEIR RELATION TO THE PREVALENCE OF BACK PAIN
Review round: 1
Reviewer: 2

Output:
Basic reporting: This article writing conforms to the writing structure of the thesis.
Line 58, "(Fig. 1, Hudetz, 2005)" is not found in the References.
Lines 246-249, "The aim of the study was to …………… and to compare the body posture of the athletes studied with their non-athlete peers." No measurement data with non-athlete peers was shown in this article.
Experimental design: Lines 111-112, “The aim of the study was to evaluate the effect of body position during play on the change in the shape of anterior-posterior spinal curvatures and trunk asymmetry in table tennis players.” This study measured the difference between table tennis player’s ready position and habitual position. There must be a difference in body structure when posture is changed. The relationship between this difference and spinal pain should be explored.
Validity of the findings: This research concluded, “after adopting this position (table tennis ready position), there are significant differences in the angles of anterior and posterior spinal curvatures compared to the habitual posture. Adopting the ready position is also associated with an increase in asymmetry in the position of the pelvis and spinous processes.” Is this a table tennis player plays with one hand and the inevitable difference in the posture change?
 Lines 55-58 , "The so-called ready position, in which the lower and upper limbs are flexed, the torso is significantly leaned forward, the rocket is kept in front of the body, the center of gravity of the body is shifted forward, the body weight is kept on the forefoot, etc. (Fig. 1, Hudetz, 2005)" This seems to indicate the possible results of this study. If this study could analyze the correlation between spinal pain and body posture of table tennis players, it would strengthen the value of this research.
    In the conclusion section, “After adopting this position, there are significant differences in the angles of anterior and posterior spinal curvatures compared to the habitual posture. This may be the cause of overloads and pain complaints reported by the study participants.” Ready position is basically relaxed and not fixed for a long time. During the game or practice, players must change different ways to hit the ball, thus changing different body positions. From this point of view, it is difficult to determine the cause of pain complaints. Is it possible that the player's high-impact actions such as smash or loop drive during the game may be the cause of pain complaints? Therefore, if the relationship between spinal pain complaints and posture differences can be found, it would be more convincing. Pain complaints may also be caused by other improper postures or weekness of related muscle, not necessarily caused by the ready position.
    Line 261, "hence the pressure on intervertebral discs increased significantly." This is only speculation based on the results of Nachemson (1987), and had not been confirmed in this study.
Additional comments: In summary, this study completed measuring the difference of table tennis player’s ready position and habitual position, and the investigation of spinal pain complaints. Although kyphotic body posture and asymmetry in the ready position  was found in table tennis players. If the study could figure out the association between spinal pain and body posture, it could improve the readability of this study. Lines 109-110, “Can overload occur in certain parts of the spine and can the asymmetry deepen as a response of adopting this position?” The results of the study did not answer the hypothetical question raised by the authors. Therefore, publication is not recommended.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HABITUAL AND READY POSITIONS IN FEMALE TABLE TENNIS PLAYERS AND THEIR RELATION TO THE PREVALENCE OF BACK PAIN
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: 1.	The title in my opinion doesn’t convey the essence of the topic, it need to be overthink and authors should try to not to use suggestive words as “asymetry” or “differency” (my recommendation eg.: Effect of playing table tennis position on posture female players).
2.	What were the sampling methods? Please complete into materials and methods/ participants part. probability or non-probability (Simple random sampling, Stratified random sampling, Systematic random sampling, Multistage random sampling, Cluster sampling/Judgement Sampling, Quota Sampling, Convenience Sampling, Extensive Sampling). There is lack of the information how the authors selected the study group
Validity of the findings: 1.	Conclusions need to be re-edited, there is not necessarily to give any explanation; my suggestion: This study demonstrated the dominance of kyphotic body posture in table tennis players., which can be caused by many hours of using the ready position during playing. After adopting this ready position, there are significant differences in the angles of anterior and posterior spinal curvatures compared to the habitual posture. This may be the cause of overloads and pain complaints reported by the study participants. Adopting the ready position is also associated with an increase in asymmetry in the position (transverse palnerotation) of the pelvis and spinous processes (frontal plane). Therefore, Training programs should be extended with exercises that relieve the spine in the vertical line and exercises that improve symmetry of the work of the upper limbs, body trunk muscles and the pelvis.
2.	line 145: who was doing the examination, marked points? it was the same person? was the person a physiotherapist or experienced researcher?
3.	line 146: in methodology of the mora there is necessary to mark also a peak of lordosis and kyphosis. does the authors marked this points also?
4.	line 178: norms by Bibrowicz (Bibrowicz 1995) are for adults or children, men or women? Please explain this or assign this fact as a limitation of the study.
5.	line 189: the player carried the racket during keeping the ready position or not? if not explain why?
6.	line 222: Explain what is the value of compensation index and on what is based this index?
7.	line 229: “moderate level” Please explain on which form was based this statement?
Additional comments: 1.	There is different English name of the school on the webside: Faculty instead department; and there is a typing error (Department of Sports, Universisty School of Physical Education in Wrocław, Wrocław, Poland).
2.	Text is not justified.
3.	Single typing or editorial errors throughout the text, please trace the text: line 174, line 137,line 176,
4.	line 29: add (averrage, sd)  age, body mass, time of study population  study population.
5.	line 30: completed a questionnaire. add a :author's own” questionnaire
6.	line 51: delete unnecessary “- -”
7.	line 68: correct “(excessive kyphosis, lordosis, scoliosis)”; my suggestion: Postural defects (excessive spine curvatures, scoliosis)
8.	line 70: even internal organs.. add “even internal organs functioning.”
9.	line 77: “muscle tears” correct into “muscle strain”
10.	line 103 and 109-111: I recommend to remove this sentence or transfer to discussion
11.	line 123: the same as in line 30
12.	line 137: “that maps the anteroposterior curvature” add a “spine”
13.	Table 1: there is no explanation for the “*” symbol. Center the text in the table.
14.	line 246-253: This is unnecessary repetition from introduction part.
15.	Authors should indicate the limitations and strengths of the study (there are some more of them except of indicated eg. Mora measurement error, sampling method; indicate the postural problems in group of table tennis players)


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HABITUAL AND READY POSITIONS IN FEMALE TABLE TENNIS PLAYERS AND THEIR RELATION TO THE PREVALENCE OF BACK PAIN
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have followed all of my corrections. The work meets all criteria for publication.
Experimental design: The authors have followed all of my corrections. The work meets all criteria for publication.
Validity of the findings: The authors have followed all of my corrections. The work meets all criteria for publication.
Additional comments: The authors have followed all of my corrections. The work meets all criteria for publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: HABITUAL AND READY POSITIONS IN FEMALE TABLE TENNIS PLAYERS AND THEIR RELATION TO THE PREVALENCE OF BACK PAIN
Review round: 2
Reviewer: 2

Output:
Basic reporting: No Comment.
Experimental design: No Comment.
Validity of the findings: No Comment.
Additional comments: Thank you for your solid corrections and explanations! It was a pleasure to do a review your paper.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMAGE DATABASE OF JAPANESE FOOD SAMPLES WITH NUTRITION INFORMATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper presents a validated image database of Japanese food items. The availability of such a database is important for cross-cultural food research. The paper is generally ell written and easy to follow.

An essential reference to an existing and freely available validate food image database that also contains many Japanese and Thai images is missing.

The raw data only contains the demographics and rating data. The food image database itself is not publicly available and will only be supplied to buyers of a “Iwasaki SAT system”, which appears to be a set of plastic food models.
Experimental design: Some details about the experimental designs are missing.
Validity of the findings: The restricted age range (only young people) and the small number of food items (only 46).  clearly limits the general validity of the results.

The variation in plateware is a serious limitation (especially when the images are to be used in fMRI studies) that should be mentioned.
Additional comments: General comments

My main concerns are (1) the fact that the photographs appear to represent replicas of plastic food items, which raises doubts about their naturalness, and (2) the fact that the image database is not made publicly available.

Abstract

Line 26: “To verify the validity of the database”: verifying implies that the validity of the database has already been established, which is not the case here. This should be:  “To validate the database”.

Line 28 and rest of the text: liking and wanting are not emotional responses, but affective appraisals

Line 32: “better” is incorrect, you probably mean that the ratings were higher

line 34: “and objective information and subjective perception of …” >> “and between objective nutrition information and subjective perception of …” >>

keywords: add valence and arousal

1. Introduction

Line 60: “existing databases contain only Western food” : this is incorrect, for instance,  the publicly available CROCUFID food image database contains images of over 200 Japanese food items  and more than 100 Thai food items (see Toet, et al., 2019, available at  https://osf.io/5jtqx/).

2.1 Participants
Some essential information is missing, like whether the participants had any color vision deficiencies, whether they were following diets or whether they had any food allergies, and their current physical state (hungry, thirsty).
Did the participants receive a financial compensation?
How were the participants recruited? From their low average age I assume that they were all students. How representative is this sample for the general Japanese population? Young people may be more familiar with Westerns food, so the difference may become more significant when a wider age range is included.

2.3 Stimuli

Lines 109-110: I understand (not completely sure since the web page to which the authors refer is only available in Japanese) that the Iwasaki food items are plastic replicas of real food. This is a serious limitation: how realistic are these images compared to real food? Why did the authors did not photograph real food items?

Line 111: The link refers to a site in Japanese; I was therefore unable to verify what it is exactly about. Please explain what the Iwasaki SAT (what does SAT stand for?) system is and why the reader should buy it to obtain the image database (line 253).

Lines 113-114: “The food samples bear a close resemblance to actual food.” >>  Please explain what these “food samples” are when they are not real food but when they only resemble food?  Why did the authors not make pictures of real Japanese food?

Line 121: Please provide the identifiers of the 5 images from the Food-pics database that were selected for this study. Why did this study include only 5 Western food images?

2.4 Procedure
Was the stimulus presentation self-paced or timed?
What was the reason to show thumbnails of all images at the start of the experiment?

3.1 Image information

Lines 113-114: “Figure 1 shows examples of images of the Japanese food samples” : it appears that all food items were plated differently. It is known that plateware significantly affects emotional and affective response to food items (Piqueras-Fiszman, et al., 2012, Piqueras-Fiszman, et al., 2013, Stewart & Goss, 2013, Van Ittersum & Wansink, 2012). This may be a confounding factor in the present study, especially since the items from the Food-pics database are not plated. Most existing food image databases therefore use a uniform or constant background. Using different backgrounds for each food item makes the images useless for brain studies, since one cannot be sure that differences in brain activity are related to differences in food characteristics or in background characteristics.

Line 159: “Table 1 lists the names and descriptions of the food” >>  please mention that this table lists all 46 items in the database

3.2 Subjective ratings
Line 166: 7 t-tests were performed, but only one (which?) result is reported for the significant differences. Also, please add the effect size.

4. Discussion

Line 195: “the images of Japanese food samples were more natural” >> Please rephrase as “the images of Japanese food samples were rated more natural”

Line 199: “than Western foods” >> This is stated too general. Please rephrase, e.g. as “than the five selected Western food items”.

Line 223: The practical significance of this database cwould increase if it was made publicly available. Now it is only offered to buyers of the Iwasaki SAT system

Line 234: The variation in plateware is a serious limitation (especially when the images are to be used in fMRI studies) that should be mentioned.

Other limitations are the restricted age range (young people), which clearly limits the general validity of the results, and the small number of food images (only 46).

5. Conclusion
Line 245: “better” is incorrect, you probably mean that the ratings were higher

Raw data:
“Fe1ale” should be “Female”

References

Piqueras-Fiszman, B., Alcaide, J., Roura, E., & Spence, C. (2012). Is it the plate or is it the food? Assessing the influence of the color (black or white) and shape of the plate on the perception of the food placed on it. Food Quality and Preference, 24 (1), 205-208. doi: 10.1016/j.foodqual.2011.08.011.
Piqueras-Fiszman, B., Giboreau, A., & Spence, C. (2013). Assessing the influence of the color of the plate on the perception of a complex food in a restaurant setting. Flavour, 2 (1), 24. doi: 10.1186/2044-7248-2-24.
Stewart, P.C., & Goss, E. (2013). Plate shape and colour interact to influence taste and quality judgments. Flavour, 2 (1), 27. doi: 10.1186/2044-7248-2-27.
Toet, A., Kaneko, D., de Kruijf, I., Ushiama, S., van Schaik, M.G., Brouwer, A.-M., Kallen, V., & van Erp, J.B.F. (2019). CROCUFID: A cross-cultural food image database for research on food elicited affective responses. Frontiers in Psychology, 10 (58). doi: 10.3389/fpsyg.2019.00058.
Van Ittersum, K., & Wansink, B. (2012). Plate size and color suggestibility: The Delboeuf Illusion’s bias on serving and eating behavior. Journal of Consumer Research, 39 (2), 215-228. doi: 10.1086/662615.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMAGE DATABASE OF JAPANESE FOOD SAMPLES WITH NUTRITION INFORMATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. English:
Overall, the English language used in the manuscript is clear, however, changes must be done in order to improve the readability of the manuscript to an international audience. In particular, the Abstract section (lines 20-39) requires major improvements in English language and concepts presentation. Stimuli section (lines 109-123) also needs major improvements since it is not clear as it is presented.

2. Literature references:
- In the Introduction section the role of visual processing in food perception is not correctly referenced (lines 43-49 in particular), important papers present in the literature are missing, such as Van der Laan et al., (2011; NeuroImage) and Huerta et al., (2014; Obesity), important meta-analysis works for the field.

- In line 48 the authors refer to “seeing annd eating food” here they should specify that seeing refers to pictorial representations (images) of food and eating refers to liquid tastants stimuli (see Huerta et al., 2014, Obesity).
Moreover, when referring to neuroimaging studies which used the abovementioned pictures databases the authors refer to a single paper, more papers must be cited here (line 58).

- The authors omitted a whole line of research on processed and unprocessed food when describing the stimuli (only processed foods in this study lines 116-117), previous findings using different techniques clearly find differences, both at the behavioral level and neural substrates, in response to these stimuli (see Aiello et al., 2018, Cortex; Coricelli et al., 2019, European Journal Neuroscience).

- When describing results on calories/fat estimations by visual inspection of stimuli (lines 74-81) the authors do not mention an important result by Horne et al. (2019, Appetite) which find that most participants were unable to accurately estimate the caloric content of most of the presented foods (images that were selected from FoodPics, FRIDa and OLAF databases). This point is relevant in light of the findings of the present manuscript (Figure 4 and S2).

3. Structure, Figures, Tables:
- Figure 1 description should contain the description of the presented stimuli, exemplar stimuli of the Western foods should be included in Figure 1

- The complete set of stimuli should be provided as Supplemental Material

- Figure 2 the dimensions of the labels of the axes (too small) and the quality of the image (bad) must be fixed in the revised version of the manuscript

- Quality of images in Figure 3 and Figure 4 is also not optimal and should be improved

- Table 1 should be revised in the description of Western foods; English description should be added (German description can be mantained but in brackets)

- Supplementary Figure 1 (S1) regression values and p values should be added to each plot

- Supplementary Figure 2 (S2) regression values and p values should be added to each plot

- A Table including the average responses of both emotional (Valence, Arousal, Liking etc) and subjective measures of nutrition (total calories, % carbo, % fat, % protein) should be added to ease the reading of the manuscript
Experimental design: 1. The main issue with the experimental design is the unbalanced number of images for Japanese (46 images) vs Western stimuli (5 images), could the authors explain such decision, since this big difference could affect all of the reported statistics.

2. Could the authors explain the small sample size (n=32) since other food images databases have larger samples (FRIDa n = 84; FoodPics n = 638 + n = 831 “german-speaking sample”; OLAF n = 612).

3. The choice of including only processed foods in the study is never explained.

4. How do the authors explain the high ratings on "Naturalness" of Japanese foods given the fact that all of the selected foods were processed foods.

5. (Lines 131-132) The authors do not explain how total calories, percentage of carbohydrate, fat and protein measures were acquired (VAS scale? n-point scale? paper-pencil? on pc?), this should be added to the Procedure section. Giving a look at the raw data a non-continuous variable was used.

6. Did the authors try to analyze the correlation coefficients between the subjective perception of nutrition and emotional responses in participants? Is there a correspondance in the results of such analysis and the reported analysis in Figure 3 (and S1) given the high correlation of objective and subjective nutrition measures (results reported in Figure 4 and S2). If of interest, this further analysis could be added in the Supplementary figures section.

7. Line 105 the image presentation was controlled by Powerpoint 2007 but the authors do not describe how emotional and subjective measures of nutrition were acquired, were the 9-point scales presented on paper-pencil or on the pc? How did the participants report the subjective measures of nutrition (see point 5)?
Validity of the findings: 1. The most interesting finding of the research is not stressed by the authors, the fact that objective and subjective measures of nutrition are highly correlated is of great interest (given the findings of Horne et al. 2019, Appetite, which find that most participants were unable to accurately estimate the caloric content of most of the presented foods), here participants are very accurate in estimating the total caloric content (very clear from figure S2), as well as carbohydrate, fat (smaller effect) and protein content.
The authors should stress such finding in the abstract and the discussion, instead of stressing the fact that previous databases did not contain objective nutrition information which is also not completely accurate given the objective values included in Western-food image databases
Additional comments: 1. Did the authors acquire information regarding participants’ hunger level at the moment of the experiment? If not, why? given the effects of hunger on explicit ratings (see Coricelli et al. 2019, Food Quality and Preference).

2. Did the authors acquire any information regarding participants’ eating habits? Were the participants all omnivores? Were vegetarians/vegans included in the study? Such info is not present in the manunscript nor in the raw data. If not, why?
Additionally, was information regarding dieting habitudes of participants acquired? given the big amount of literature and the well-known effects of dieting (Restrained eaters) on explicit ratings (see Hoefling & Strack, 2008, Appetite and Coricelli et al. 2019, Food Quality and Preference).

3. Low-level objective measures of the images are not reported in the manuscript (i.e. brightness, spatial frequency, image complexity), why?

Minor changes:

- Line 32 replace the term “better” with another term more specific to the results.

- Line 43 replace the term “controlling eating behaviors” with a more exhaustive explanation of the role of visual processing in food perception such as the ability to extract inherent information of the presented foods (i.e. edibility, Tsourides et al., 2018, NeuroImage).

- Paragraph from 224-233 must be revised; both objective (caloric/fat, healthiness, level of processing) measures and subjective measures (ratings) have been investigated with EEG (see Toepel et al., 2009, NeuroImage; Bielser et al. 2015, Brain and Cognition) and fMRI (see Killgore et al., 2003, NeuroImage; Frank et al., 2010, Brain Research).


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMAGE DATABASE OF JAPANESE FOOD SAMPLES WITH NUTRITION INFORMATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors did a nice job in revising the manuscript, and appropriately responded to all remarks of the reviewers.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMAGE DATABASE OF JAPANESE FOOD SAMPLES WITH NUTRITION INFORMATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Overall there was an improvement in the revised version of the manuscript, in particular in the Introduction section and the paragraph of the Discussion assessing the limitations of the study. The complete set of stimuli in the Supplemental Figure 1 eases the reader to understand the type of food stimuli chosen. The additional results regarding the low-level features (i.e. brightness, sp. frequency, entropy) comprising three Tables of results should be moved to the Supplemental material.
Below specific modifications to the text that should be implemented.

- Line 35 I suggest replacing “the existing database” with “an existing database”

- Line 192 I suggest replacing the term “physical” with “medical”

- Line 198 I suggest replacing the term “visual deficiencies” with the term “color vision deficiencies”

- Line 268 "Participants" should be in lower case

- The authors must make sure they were coherent with the terminology throughout the paper, in particular with the terms “subjective affective ratings” “subjective appraisals” and “subjective evaluations”, (i.e. lines 157-162, 322-326). One term should be chosen and used in the entire manuscript/figures/tables and captions.

- Line 464 “are valid visual stimuli of Japan foods” consider revising this sentence

- Lines 519-521 “Use of the current database may allow studies of the specific brain activities associated with a visual analysis of food nutrition.” This sentence must be reformulated (or removed). Visual processing of food stimuli is quite consistent across studies as shown by the results of meta-analyses and reviews of neuroimaging data (correctly cited in the Introduction), the proposed database contains most of the information contained in other image databases therefore it would not reveal “new” brain activities of visual analysis of food stimuli. It also presents plastic representations of foods, factor that must be taken into account in comparing the visual processing of such stimuli.

- Line 554 I suggest replacing “our stimuli contained different plates” with “our food stimuli were depicted in different plates”

- Line 566-567 “restricted participants” remove “restricted”

- Results related to the low-level features should be moved to the Supplemental material.
I propose to move Table 6, Table 7 and Table 8 to Supplemental material.
Moreover, Table 7 and Table 8 should have a consistent order of statistics (t, p and r or r, t, p)


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: LEGACY EFFECTS OF HISTORICAL GRAZING ALTER LEAF STOMATAL CHARACTERISTICS IN PROGENY PLANTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Poor English.
Experimental design: Poor design.
Validity of the findings: Not sure
Additional comments: My comments are as follows:
1.	For the legacy effect, it is important to compare the phenotypes between parent plants and progeny plants, not the phenotypes between the grazed plants and CK. I did not see any information about the stomatal characteristics of parent plants in both grazed and fenced pastures. Therefore, it is not clear whether it is due to genetic performances of the plants or not. I suggested to add another experiment in the field to test the difference of stomatal characteristics of grazed plants and the CK. If it shows the same phenomena as the greenhouse experiments on bud. Then, you can say this is the legacy effects.
2.	For the grazing experiment, I am not sure how you can define overgrazing? What is the criteria to rule the overgrazing in your study.
3.	I am not clear what the linkages between the 8 genes measured and stomatal characteristics. You need to make clear frameworks/flowchart for the relationship between genes and the stomatal characteristics.
4.	How can you draw the conclusion of down-regulated 2-18 times and up-regulated 3 times without any quantified analysis?
5.	English is poor. There are many grammatical errors in the paper. Please find a native English speaker to make corrections.