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: DNA BARCODING FOR BIODIVERSITY ASSESSMENT: CROATIAN STONEFLIES (INSECTA: PLECOPTERA)
Review round: 1
Reviewer: 3

Output:
Basic reporting: Wow, this was a tremendous amount of work. I commend you on this ambitious undertaking. You have created a resource that will be used far into the future and you have built a professional roadmap for yourself and others to improve our understanding of the taxonomy and conservation ecology of one the most environmentally sensitive of aquatic groups in the world. Your approach is the route forward for Plecoptera taxonomy and systematics, and your work has application to conservation of aquatic systems as well. I look forward to great work from you.

The English in places is a bit rough. I have tried to suggest improvements. There are some places that I said I did not understand and state so. I think your prose would benefit from writing in the in the first person as opposed to passive voice.

You have covered the literature well, though I have suggested some North American literature that is pertinent.
The article is professionally done. The analyses are thorough. It is a totally original work.
Experimental design: Well done throughout. This is how I want to see taxonomists enhance their good work with molecular approaches.

Questions are well-defined. This work certainly fills a gap in the knowledge of the distribution and genetic variation of species and contributes a roadmap for improving the taxonomy of one of the most environmentally sensitive aquatic groups in the world.

Methods are well documented and should be a model for biodiversity work.
Validity of the findings: Findings are impactful and novel. The work challenges the field of Plecoptera taxonomy to improve in the use of molecular data to circumscribe species and test hypotheses of distinctness of species.

The underlying data have been provided in myriad ways. Analyses are sound and varied in order to test for consensus.

Conclusions are sound and support results.
Additional comments: Some improvement in English is necessary but should not be difficult. I always markup manuscripts to the extreme. Use what you can.

In the results section, I had some difficulty understanding what you were trying to do. I thought you might want to separate the morphological from the molecular results somewhat. I think you might want a Table 1 of species found that is not supplementary. I had some difficulty with your counts of species, so I looked at your specimens supplementary table and could get the numbers close to what you state in the results. It was not until I looked closely at Figure 2 that I saw exactly where 74 species came. You did a great job explaining that circular gene tree.

I don't know what the journal wants, but you don't usually provide author and year with species names, but in some places you did. Be consistent.

You present in some place proportions for genetic difference, others %, but the figures are in %. I expected they would be the same. % is easier to understand in text.

Figures 1 B & C are not particularly informative. The horizontal scale is hardly used to accommodate the data. 0-10% seems to be the appropriate data range . I also thought they might be better as histograms.
Figure 4. Some dots on this map are larger than others. You have richness color-coded, but there is no legend for size of dots.

You frequently talk about "obtained sequences". I think you try to distinguish between your sequences and those from BOLD or GenBank. Maybe here is a good place to use active, first person to shorten the sentence and be more direct, e.g., "Our sequences".

The Discussion is a little long. They seem to be a few to many asides in the text. Stick to the most important information in an effort to shorten the section. Also, in this section you begin a lot of sentences with an abbreviated genus name. I don't think that it is good practice to begin a sentence with an abbreviation and I try not to do it with an acronym either.

In relation to species you occasionally write "source areas" in relation to habitat. I don't know what you mean. Maybe spring sources or headwaters is you intent. If so, say that.

A few citation practices bothered me a bit. First, you citing my Plecoptera Species File for narrow distribution of a species or group is not appropriate and it robs your colleagues of a citation of their work that is specific to your needs. You also cite unpublished data a bit too much. Cite it when it is published--I expect you will have plenty of papers in the future. Also, you need to look at references that are broader than Europe. I have provided several North American references that should be specific to your needs.

Line 618. Species occur at elevations, not altitudes.
Line 692. Perla burmeisteriana Claassen, 1936 is the valid name for the homonym Perla abdominalis Burmeister, 1839. Perla abdominalis Guérin-Méneville, 1838 is also valid, the first instance of Perla abdominalis, by one year.

Throughout your subsection "Extinction and conservation" you use the term extinction to mean the global and local loss of a species. Please use the term extirpation for local loss or range loss.

Supplemental Table S1 has a few errors or suggested changes needed.
I would like to see a column for Family added. It will help others more effectively sort the data.
You record two variants of Leuctra hippopus, most with 1899 year of description, but one incorrect one with 1900.
You list a Leuctra sp. Z, is this correct? It appear nowhere else.
Isoperla popijaci does not have authority or year of description.
Taeniopteryx sp. has not temporary designation. I think that might be useful to you in the future. All are from Croatia currently. Maybe name it "Taeniopteryx n.sp.CR-1"


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: CHANGES IN SERUM AMINO ACID LEVELS IN NON-SMALL CELL LUNG CANCER: A CASE-CONTROL STUDY IN CHINESE POPULATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this study, the authors evidenced that the level of several amino acids were modified in Chinese NSCLC patients compared to control patients. One amino acid was also expressed differentially depending on the subtype of NSCLC (ADC or SSC). While the alteration of AAs in serum is well established, no study reported such effects in a specific population such as Chinese patients and never by considering subgroups of ADC and SSC. This study is interesting and the article is well written and respects the standards for an original research article. Nevertheless, several aspects mentioned in the comments below should be clarified to consider this manuscript for publication
Experimental design: In this study, the authors quantified the level of amino acids in serum to evidence that the level of several amino acids was modified in Chinese NSCLC patients compared to control patients and that one amino acid was also expressed differentially depending on the subtype of NSCLC (ADC or SSC). While the method used is well appropriate for such aim (in term of number of patients included in the cohort, accuracy of the quantification method, statistical approach) and several important details are well described, it would be helpful to edit the manuscript according to the comments below
- line 136, the authors justify the choice of using Kolmogorov–Smirnov test rather than other statistical multivariate tests
- line 141: “a validation set with a ratio of 7:3.” The authors should specify what is this ratio
- Line 178” After adjustment, the concentration of Trp was significantly decreased,” The authors should explain what are these adjustments.
- Line 182-190: The authors consider only the P-FDR lower than 0.01, but the ratio of AA level was pretty close to 1 (i.e, 1.07). Is it relevant to consider AAs with such ratio as significantly deregulated? In my point of view, it would be more correct to apply a threshold a ratio from which the AAs is considered significantly deregulated with a P-FDR lower than 0.01
- Line 247: It is well known that the nutrition impacts the AA expression levels. So, I was wondering the patients followed a specific diet or at least if those data where recorded to ensure that the difference of expression is link to NSCLC and to different diets.
Validity of the findings: Through this study, the authors evidenced modulations of several AA expression level in serum of Chinesse patients to discriminate the NSCLC patients to control patients. They also evidence that Asp expression change depending on the subtype of the NSCLC (ADC or SSC) It will be helpful to classify teh control from NCSCL cancer,but the auhtors should answer the comments below to improve the manuscript

- Table 1. I have concerns that the difference between control and cases groups is significant regarding smoking and BMI aspects. This may have impacts in AA expression level and interferes in the results of the study.
- The authors should present a table of clinical characteristic for the classification of subgroups as they did in table 1
- the authors should discuss about the interest of their method campared to the methods that are currently used in clinics
Additional comments: 1)	An abbreviation list would be helpful to read the article such as for EAA, NEAA, IQR
2)	Line 68: The authors should also add that several AAs such as CIT also activate the mTOR1 signalling pathway through the modulation of the concentration of PA and Rheb activity in the LEL by citing, for instance, the review S. Bourgoin-Voillard et al Proteomics 2016, 16, 831–846
3)	Line 187-190: “However, except for the higher Asp in SCC (P-FDR<0.01), there was no significant difference in AA levels between adenocarcinoma and SCC.” The comparison of AA expression level in subgroups is not shown for all AAs presented in the article. It would be interesting to present it to all AAs and not on a few of them. Otherwise the authors should justify why they selected those AAs.
4)	Table 2: As CIT seems deregulated, it should be in bold
5)	Line 249-262: the authors should also discuss about the role of CIT in mTOR signaling pathway activation
6)	It would be interesting to establish a ROC curve by considering not only one AAs, but several AAs. Such combination may increase the specificity of the biomarker test


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: CHANGES IN SERUM AMINO ACID LEVELS IN NON-SMALL CELL LUNG CANCER: A CASE-CONTROL STUDY IN CHINESE POPULATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper is intriguing and well-written, offering insights into the topic of cancer. I only have two small points to make.
Experimental design: Please provide the internal standard utilized to determine AA.
Please provide further information for metabolic pathway analysis.
Please provide a power analysis of your study as well as type-II error statistics.
Validity of the findings: Please provide a power analysis of your study as well as type-II error statistics.
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: CHANGES IN SERUM AMINO ACID LEVELS IN NON-SMALL CELL LUNG CANCER: A CASE-CONTROL STUDY IN CHINESE POPULATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors improved properly the manuscript according to my previous comments. The manuscript is thus, ready for publication.
Experimental design: no comment
Validity of the findings: no comment
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: CHANGES IN SERUM AMINO ACID LEVELS IN NON-SMALL CELL LUNG CANCER: A CASE-CONTROL STUDY IN CHINESE POPULATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript is well written and ready for publication.
Experimental design: Experimental methods are well designed.
Validity of the findings: The findings are validated using training 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: CHANGE OF INTESTINAL MICROBIOTA IN MICE MODEL OF BRONCHOPULMONARY DYSPLASIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: 1. Write HE staining as hematoxylin and eosin (H&E), if it appears for the first time in the manuscript.
2. For the induction of BPD neonatal mice were subject to high concentration of oxygen (80±5%) for 3 weeks. How did the authors reach to these figures. What about other concentrations?
3. Mention the mice species in the M & M
4. The authors are suggested to submit the data to appropriate data bank and mention the accession number.
5. Validation with human subject is suggested.
Validity of the findings: 1. How can we reach to the species level with a single taxonomic marker i.e., 16S ? Why microbiota study upto species level was not performed?
2. There is no mention of fungal diversity.
3. Justify, how the experiments done in animal model can be used to validate the diversity of microbial diversity in human infants.
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: CHANGE OF INTESTINAL MICROBIOTA IN MICE MODEL OF BRONCHOPULMONARY DYSPLASIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. The disease in question is bronchopulmonary dysplasia. There is no hyphen.
2. Its Linear discriminant analysis not line discriminant analysis.
3. The paper completely fails to discuss the relevant literature. Since all of these analysis have previously been published in both humans and mice, basic scientific practice would be to discuss this in the introduction and discussion, as this is the point of those sections of a scientific paper. Some examples by pubmed ID: PMID: 31644311, PMID: 32877224, PMID: 31644312, PMID: 27488092, PMID: 32801143, PMID: 31664856, PMID: 32117822, PMID: 30487069, PMID: 31967848, PMID: 31898918.
4. The microbiome data in the tables is of limited value and should likely be placed in the supplement.
Experimental design: There are significant limitations in reporting of rigorous research and in study design.
1. Since this study replicates a portion of this study in AJP Lung (PMID: 31644311), I would suggest starting there to see the level of detail needed in reporting the model design and microbiome analyses.
2. The most significant issue is in statistical power. In neonatal microbiome studies the biological replicate is the Litter, not the individual mouse as all pups in the same litter are effectively technical replicates. Therefore the effective samples size is 1-2 for most of these experiments. The whole paper needs to be repeated with markedly more samples. PMID: 32688019
3. In figure 1, the representative histology does not appear to have been prepared correctly as the samples appear under inflated, this calls into question whether the authors actually performed the hyperoxia exposure as described. Since this model is well published norms are available to quantify these values. I suggest following the procedures in the ATS guidelines (PMID: 23624789) and the norms in the paper by the Morty lab: PMID: 29315540.
Validity of the findings: 1.Validity is uncertain as the study is vastly underpowered - see point 2 in Experimental Design
2. Sequencing data must be uploaded to a public repository.
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: CHANGE OF INTESTINAL MICROBIOTA IN MICE MODEL OF BRONCHOPULMONARY DYSPLASIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: This is a nice experimental paper investigating the importance of the gut-lung axis microbiota in the development of BPD. The paper is well designed, nonetheless there are important issues, mainly in the statistical power, that make the paper unsuitable to be published in the present form.

English is overall acceptable, but sometimes the grammar and construction of the phrases makes it hard for the reader ti follow the reasoning or the meaning of the sentence. For example:
"Compared with the control group, the development of lung tissue in model group has been destroyed. " what does this sentence mean?
"The  microflora in gut and lung are rather complicated, which play important roles in the pathogenesis
of pulmonary and intestinal diseases through the gut-lung axis": the sentence is not clear, please rephrase
Line 255: "alternation" Do the authors mean "alteration"?
Line 258: "In the presence of neonatal maturity, the gut microbiota gradually stabilized."  the authors mean " with the progression of neonatal maturity..."?

Therefore the paper has to be revised by a native speaker to improve the quality of the writing and the reasoning

Literature references are overall sufficient, but as the authors are referring to a BPD model , the "new BPD"  has to be cited. In fact the authors refer to the development of BPD related to ventilation and oxygen toxicity, but they have to cite also the changes in vascular structure !that are now the main cause of BPD in extremely preterm neonates.   (Bronchopulmonary Dysplasia: 50 Years after the Original Description.  Bancalari E, Jain D. Neonatology. 2019;115(4):384-391. doi: 10.1159/000497422. Epub 2019 Apr 11.  PMID: 30974430)

Raw data are correct tell shared
Experimental design: The most significant issue is the statistical power. the experiments have been carried out with a limited amount off samples (5 for each group)  Therefore the results have not enough statistical power to be accepted for publication. The experiments should be repeated with more samples as to confirm the results obtained so far. The paper could be accepted as preliminary data foreseeing a more extensive analysis.

Ethical standards were correctly met.

Methods were correctly described.
Validity of the findings: As commented previously, the main issue is the insufficient sample size.

Sometimes data are confusing and contradictory: for example line 195: "there was no significant difference in intestinal microbiota between two groups" opposed to line 200 "and the differences in gut microbiota between two groups were statistically significant (P<0.05)."

Is is not clear if some of the findings discussed in the paper were analyzed in the paper or are just referred from other paper (for example "There was massive generation of TNF-α, IL-6, IL-8 and MCP-1 in lung tissues(Bhandari 2010) " was this confirm by the paper or just cited?)

The authors found 3 different metabolic pathways: Signal transduction, glycan biosynthesis and metabolism , metabolism of terpenoids and polyketides. The authors should extensively comment on the importance of these pathways in the discussion.
Additional comments: With the proposal of the gut-lung axis concept(Budden et al. 2017).: the sentence has to be completed

Line 237 "As previously described, microorganism would affect the progress of certain diseases rather than improving their severity" what do the authors exactly mean? Please explain better

Conclusion:
the is no need to repeat the experimental model her "A BPD model was established in neonatal mice through exposure of hyperoxic environment. Then we analyzed the difference of gut microbiota diversity between natural mice and BPD model mice"

The authors should comment on the potential importance of probiotics to change the intestinal microbiota and reduce the incidence of BPD.


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: CHANGE OF INTESTINAL MICROBIOTA IN MICE MODEL OF BRONCHOPULMONARY DYSPLASIA
Review round: 3
Reviewer: 1

Output:
Basic reporting: the reviewers addressed my concerns in the new version of the paper
Experimental design: the experimental design is clear
Validity of the findings: my concerns on number of the samples have been cleared
Additional comments: the reviewers addressed my concerns in the new version of the 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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper reports the results clearly and in sufficient detail. Clearly written throughout. Introduction shows context clearly. Raw data has been supplied.
Experimental design: This study appears well design to answer the research question, though it is cross sectional and a longitudinal design would have been preferable. This is primary research in line with the journal scope with a clear research question. Ethics approval and considerations is outlined and enough detail is given for replication.
Validity of the findings: Findings are clearly presented, statistics seem appropriate and demographic variables are controlled for. It’s finding about cash reserves being more important than financial shocks is relatively unique an important one for the literature. Data has been provided and the statistical analyses as far as I can tell are sound (I believe they used a difference software package from me so I cannot repeat the analysis but they appear sound from the details given). Conclusions are clear.
Additional comments: •	Introduction line 68: ‘habits in saving’ please reword e.g. ‘usual habits in saving’.
•	Introduction line 77: Change ‘leaky’ to ‘leaking’.
•	Materials line 108: Please explain why the US amount for emergencies of 400 dollars was used and then this change to Thai Baht: Was there not such an estimate available for Thailand specifically to use as a benchmark for case available for emergencies?
•	Materials line 128: Please add to paper that the measure of financial stress by Siapush has also been used in previous studies of mental health and finances (e.g. Richardson et al., 2007).
•	Materials line 136: Please clarify if the PHQ and GAD measures have been formally validated with a Thai sample and if so provide a reference. Please also provide the chronbachs alpha of the GAD-7 in this sample to demonstrate reliability of this measure. Please also clarify how this measure was used in the analysis: I am unclear if this was used in the results i.e. using this score to determine if they had a financial shock. There are other analyses which could have been conducted e.g. correlating PHQ and GAD depression scores with scores on this financial measure: Please explain why this was not done and clarify how this financial measure was analysed in the data.
•	Materials line 177: It says that missing data was excluded ‘except for questions on sensitive issues’, please justify this and explain in more detail: What constituted a sensitive question? Why was this not excluded, and what happened with this missing data?
•	Materials and method line 184: Please clarify what statistical software was used for the analysis.
•	Results line 220-221: Please remove ‘(n=1,555 participants)’ as this is repeated twice in this sentence.
•	Results line 226: Please add the % to ‘the majority of participants’. I am also not sure if you could describe 51.7% as majority of participants unless you conducted a statistical analysis to show if this gender difference was statistically greater than expected.
•	Results line 230 ‘slightly higher than those’ add % here as well please.
•	Discussion line 265-272: I do not think this much discussion about prevalence of credit card use in Thailand is needed, it is not especially relevant to the results. Please edit down.
•	Discussion line 293 onwards. Please add to discussion about limitations that perhaps a more global measure of stress could have been included. I think it is also important to place more emphasis on the cross-sectional nature of the study as a limitation: Reverse causality cannot be ruled out e.g. it could be that those who are prone to depression are less likely to have savings rather than not having savings increase the risk of experiencing depression symptoms.
•	Discussion lines 316-323: There is a lot of discussion about the link between finances and binge drinking, including you saying that ‘those who experienced economic distress were less likely to self-report binge drinking’. Please correct this and edit down this section: There was a non-significant trend, so this needs to be clarified in the discussion and I don’t think this must discussion about alcohol us warranted giving a non-significant finding.
•	Discussion line 324: You say qualitative methods should be used I think longitudinal analysis would be more helpful to look at the link between covid financial impact and binge drinking.
•	Discussion line 330: You say that the characteristics of your sample ‘closely reflect’ the demographics of the Thai population. How can you be sure unless you have formally compared the demographics in the sample? I don’t think such a full formal analysis is required, but linking here to some data about broader Thai population could help with a brief analysis e.g. gender break down and average age here was X compared to general population.
•	Discussion line 337: Add to future studies about the importance of longitudinal studies in the area.


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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: A summary of research
Employing a nationally representative telephone-based survey in late April 2021 with a sample size of 1,555 participants, this study assessed the extent that mental health outcomes and health behaviours were associated with the availability of emergency cash reserves and experience of economic distress in the general adult population in Thailand. The authors found that participants who experienced economic distress but had emergency cash reserves had a lower prevalence of anxiety, depressive symptoms, and a history of gambling than participants who did not experience economic distress but had no emergency cash reserve.

Comments
Abstract
1.	In the Background, it makes more sense to link the importance of emergency cash reserve to economic problems caused by the COVID19 pandemic rather than link it directly to the COVID19 pandemic.
2.	It seems the Cis in Table 2 do not correspond to the Conclusion. The CIs of the two mentioned categories are overlapped.
Methods
3.	The Sampling Methods should be put close to the Study Population and Sample Size Calculation.
4.	The study population should be spelled out clearly whether they are people who have household phone numbers or not. Please make it clear that the phone number list covers the cell phone or household phone. Please provide information regarding the coverage of Thai people who have phone numbers used in the study, and how well the people who have phone numbers represent the general population. The poor people who do not have any phone number may get worse economic and psychological impacts from the pandemic as compared to those who have one. The authors mentioned the sampling list and the replacement sampling model are belong to the research firm; however, this information should be described adequately.
5.	Please explain more about the sampling lists of both sampling levels. For example, the first sampling level uses the individual list or household list; the second level uses what phone number list. Please give more detail on what cumulative systematic sampling is for both sampling levels.
6.	Please provide a bit of information describing the quality of the field researchers (e.g., university students or the research company’s regular researchers) in order for the reader could assess the quality of the study data.
Results
7.	Line 218, please consider changing the term ‘the participant rate’ to ‘the response rate.’
8.	Line 243, please change ‘or’ to ‘for.’
9.	It would be good to show the ORs for all controlled variables in order to know how much those variables influence the outcomes.
10.	The reader might understand all parameters easier by 95%CI than S.E.
11.	It would be good to do cross-tabulations between ‘personal monthly income’ and several other variables, including credit card use, emergency cash reserve, the experience of economic distress, and all health outcomes. These tabulations could show how the baseline income is important and in what dimensions. Or, to do the current analyses but stratified by personal income because it could illustrate the effect of personal income on the outcome measures.
Discussion
12.	The authors should compare the sample characteristics with the Thai population characteristics to explicitly show how well the sample represents the population. Do not have to do an explicit table.
13.	The authors might consider using a different classification for identifying the ‘emergency expense variable’ in the analysis. For example, treating those who use a credit card as ones who have no cash may cause misclassification bias because using a credit card may not mean they have no cash. These people may use their credit cards because it is a better option for them (e.g., save cash to use for other things, no interest, get points, etc.; poor people could not have a credit card).
14.	It might be better to demonstrate sensitivity analysis of the effect of a different definition of the cash reserve than to explain that there was a small prevalence of those who used credit cards in this study.
Additional comments: no comment


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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors address an interesting and important topic related to the relationship between an individual's financial situation and their experience of anxiety and depression. In reading the paper, I am not entirely sure of their conceptual argument or the exact research question they are attempting to answer. Lines 25 to 27 present the objective as assessing "the extent to which mental health outcomes and health behaviors were associated with the availability of emergency cash reserves and experience of economic distress." Lines 84 to 87 present the objective as assessing "the proportion of the general adult population of Thailand with cash reserves for a small emergency, and the extent that the availability of such emergency cash reserves was associated with mental health and behavioral health outcomes during the COVID-19 pandemic." Lines 252 to 255 present the objective as describing "the availability of emergency cash reserves . . . and experience of economic distress uring the COVID-19 pandemic, and their associations with mental health and behavioral health outcomes." The analysis examines the interaction of economic distress and emergency cash reserves although the authors do not discuss moderation.

It seems that a conceptual model and explicitly stated research question might be helpful in clarifying the objective of the study. From my reading, it seems that the question is whether the relationship between the experience of economic distress and mental health depends on the individual's level of emergency cash reserves. For example, the authors state that "participants who experienced economic distress but had emergency reserve had lower prevalence of anxiety, depression, . . . . " (Lines 45-46) This is just a guess, however, and might be incorrect.

Abstract:
It would be helpful to have the mode of survey administration noted in the methods section of the abstract.

Introduction:
The authors make several interesting points in the introduction but do not tie those points together in a clear conceptual argument. There are several leaps of logic. For example, lines 68 and 69 read: "habits in saving for emergencies were less likely to experience economic hardship (Gjertson, 2016). In this regard, maintenance of cash reserve for emergencies may help to lower the likelihood of negative mental and behavioral health outcomes . . . . " Gjertson was examining the relationship between having emergency savings and the experience of economic distress (material hardship). How does her study support a hypothesized relationship between cash reserve and mental health outcomes? The use of the Gjertson article would suggest that the authors are arguing that economic distress mediates the relationship between cash reserves and mental health outcomes?

Lines 60 and 61 state: "Identification of other factors associated with depression, anxiety, and other behavioral health outcomes thus have implications for public health during this pandemic." This statement appears to be related to the intended contribution of the research. It would benefit from an explanation of the implications. Why is identifying other factors important? Relatedly, lines 83 and 84 state: "Empirical data on such prevalence and associations can provide useful basic information for stakeholders in economics and behavioral health." Are there examples the authors can provide? Is there an explanation of why such individuals need this basic information?

There are also several typographical errors and some spacing issues in this section and throughout the paper.

The reader would benefit from an explanation of the behavioral health disorders (gambling, drinking, smoking). No literature was presented to support the inclusion of these outcomes. No attempt was made to explain why mental health (anxiety and depression) and behavioral health (gambling, drinking, smoking) were both included or how they relate to each other.

No hypotheses or research questions were presented for either outcome.

Materials and Methods:
This section is second in length to the discussion (about 30% of the paper). There is tremendous detail related to the data collection methods, training the data collection staff, etc. It seemed a little disorganized as some of this information was presented in study design and setting. Then again in sampling methods, data collection and quality control, monitoring and data cleaning, and ethical considerations. While these subsections have different purposes, those purposes could be made a little clearer. I think that might help the reader appreciate what was done and why it is important for them to know.

In the methods section, two things are sprung on the reader without advance notice in the introduction. One is the behavioral health outcomes and the other is the past 30 day economic distress. These constructs should be introduced in your introduction and incorporated into your conceptual argument.

Were the data collection teams assigned to a region? Or did they all work across regions? If assigned to a region, how did you control or adjust for that?

How did you identify your weights? What adjustments were made to address the complex survey?

Results:
I was expecting a much more complete explanation of your findings. Other than the more robust description of findings related to sample characteristics, this section did not provide more detail than the abstract. I was also expecting to see the details of your logistic regression. Where is your pseudo-R-square? -2LL? Percent classified correctly? You have created an interaction between economic distress and emergency cash reserves yet you do not present the results of the interaction. You also do not consider main effects of economic distress and emergency cash reserves. These details would help your reader understand what you found and set them up to understand what it means in your discussion. Having one or more research questions would help you present the results in a more intentional way.

Discussion:
The discussion would also benefit from having research question(s).

The organization of this section could be improved to support the reader's ability to digest the points being made. For example, the authors discuss limitations of the measures beginning on line 294. Then they present limitations again starting on line 327.

Lines 294 to 300 should be examined. It is difficult to understand the point(s) being made in the current language.

Conclusion:
The conclusion re-presents the findings rather than providing a summary or conclusion for the paper.

Other than the typographic errors, the spacing issues and a few lines, the paper is written clearly.
Experimental design: This paper reports on a cross-sectional study rather than an experiment.

Additional explanation of the methods and alignment between the methods, research question, and results would be helpful. What the authors describe as their method: "We used multivariate logistic regression models to assess the extent that the prevalence of the outcomes differed between each population groups with adjustments for experience of economic distress within 30 days prior to the survey (if applicable) and demographic and socioeconomic characteristics" (lines 187 to 190); is not derived from a research questions or related to what was presented in the results.

There is no research question.The gap and the contribution (intended or actual) are not clear.
Validity of the findings: There are no research questions.

Presentation of the findings seems rushed and limited. The discussion has considerable detail but does not draw important conclusions (perhaps due to the lack of a clear research question).
Additional comments: The paper has potential. The points I have raised are more related to the presentation of a conceptual argument, methods, results, etc. than to the underlying data.


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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 2
Reviewer: 1

Output:
Basic reporting: See initial review
Experimental design: See initial review
Validity of the findings: See initial review
Additional comments: See initial review


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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: It would be easier for the reader if Table 3's title is more specific: showing the alternative definition explicitly in the title.


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: MODIFICATION OF THE ASSOCIATION BETWEEN EXPERIENCE OF ECONOMIC DISTRESS DURING THE COVID-19 PANDEMIC AND BEHAVIORAL HEALTH OUTCOMES BY AVAILABILITY OF EMERGENCY CASH RESERVES: FINDINGS FROM A NATIONALLY-REPRESENTATIVE SURVEY IN THAILAND
Review round: 2
Reviewer: 3

Output:
Basic reporting: The paper still lacks three journal standards:
Literature references, sufficient field background/context provided.
Professional article structure
Self-contained with relevant results to hypotheses

See attached document for comments
Experimental design: The research objective is clearer but the research question is not well defined. The authors do not make a strong case as to why or how the question is relevant or meaningful. There is not an identified knowledge gap.
There are gaps in the method description.

See comments in attached document.
Validity of the findings: It is difficult to assess the conclusions due to the way in which the discussion/conclusions are structured as well as the overall structure of the paper.

See attached document for comments
Additional comments: See attached document.


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: KNOWLEDGE OF ACUTE STROKE MANAGEMENT AND THE PREDICTORS AMONG MALAYSIAN HEALTHCARE PROFESSIONALS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: Can you explain the setting of healthcare system in Malaysia under study settings/design. The difference between primary and hospital care, what’s their functions and personnel?
How many district hospitals, university hospitals were involved? How many hospitals that have stroke units were involved? What kind of specialists/physicians were involved, from which departments were residents involved from? How many private vs the government hospitals were involved?
Validity of the findings: No comment
Additional comments: This is a good and relevant article since stroke is one of the leading causes of disability and mortality in low- and middle-income countries. Despite this, there are some clarifications that are needed.
1. Different healthcare professionals have different roles under stroke management. For example, doctors are responsible for neurological examinations, interpretation of CT/MRI images, and initiating thrombolysis after excluding contraindications, while nurses nurse patients, therefore they do not have the same roles. Therefore, it is difficult to compare them.

2. Looking at the general knowledge of stroke management questionnaire (GSK) and some other few questions are the only parts where one can compare different healthcare professionals since it deals with among others stroke symptoms which are crucial for everyone. Otherwise, other parts of the questionnaire are specific for specific healthcare cadres.

3. Under discussion, some parts of the questionnaire (e.g., stroke symptoms, thrombolytic therapy window, etc) can be compared to some previous studies. Compare with some previous studies that have assessed similar parts, e.g.,
Yang J, Zhang J, Ou S, Wang N, Wang J. Knowledge of Community General Practitioners and Nurses on Pre-Hospital Stroke Preven tion and Treatment in Chongqing, China. PLOS ONE 2015; 10: e0138476.

4. Under limitations: Not all questions on stroke management were included in the questionnaire e.g., thrombolytic medication dosages, contraindications, etc
5. Results - Factors associated with knowledge: Most of the overall knowledge of stroke findings are results of the subsections (GSK, ASM) such that they could be incorporated under it.


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: KNOWLEDGE OF ACUTE STROKE MANAGEMENT AND THE PREDICTORS AMONG MALAYSIAN HEALTHCARE PROFESSIONALS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 69: Rose over what timeframe?
73-74: Change to “Healthcare professionals’ related…” No hyphen needed
76: Change to “possibly” or “likely contributing”
86: Comma after [7]
107: Comma before and
112: For readability, add “The” at the start of the sentence.
113: Change to a :
115: Wording with transition between ASM and Cronbach’s alpha is confusing. The survey as a whole has the alpha level described or just that section? Perhaps two sentences.
115: Replace “They” with “The items” or something descriptive vs the pronoun.
230: Reword sentence about gender differences
124: Although scores are plural, the noun is variable so should be “was categorized” as it is currently written. Or adjust the sentence to flip the wording of Knowledge scores were categorized…, which were utilized as the outcome variable.
262-263: Wording is awkward. The patient isn’t being lowered, but their elevated BP is. Therefore, reword to “Elevated BP after a stroke should carefully be lowered at …”
264: Needing a verb. “…paramedics were not managing”
279: on the front line
283: answering incorrectly
Experimental design: Experimental Design
Strengths: Research question well defined as was the gap in the literature to guide this study.
87: Good job identifying a gap in the literature to form the foundation of the study
116: Perhaps introduce this chapter with the two descriptions of the online survey, and then as section 2 is being described, include the information that is currently 112-115.
354: According to the start of the methods section, the online course mailing list was used to recruit subjects. Is it known if the course had been completed? Or is there a measurement tool (pre/post) for the effectiveness of the training? If all participants have taken the training, it would be anticipated that the knowledge is higher than other medical professionals. Likewise, in the limitation section, if the participants were part of a stroke education mailing list, this would likely bias the results as discussed. This is also an opportunity for replication by using a different means to sample a larger representation of the population that may have even less experience with people with strokes.
Validity of the findings: Validity of Findings
Strengths:
It is important to see that even with practitioners that are seeing patients with strokes at times and have registered for education, still are lacking in areas of advanced stroke management. This has implications for a broader sample that isn’t recruited from a stroke mailing list. A follow-up or replication study could investigate practitioners with wider backgrounds. The findings also reinforce the need for continued education, especially for individuals who do not regularly treat people with stroke. Getting appropriate healthcare fast is key and knowing where to find those resources quickly is of utmost importance.
248: Why would a negative response be preferred with GSK-10? Is it linked to that previous education was happening but changes in performance were not being reflected in the workplace? For practitioners who are not regularly treating patients with stroke, it seems regular education would be beneficial. This may involve including discussion from the initial articles from creating the tool in describing this question. Line 256 does a nice job explaining why the negative answer is correct for GSK-8, this would be helpful for the GSK-10 segment. Line 350 states that training is a strategy to improve quality of care, which seems to contradict this test item.
197 (and throughout): Replace insignificant with not statistically significant or not statistically significant findings because being not significant can often be important.
268: 14% of GPs?
287: Good discussion again on why the item is incorrect with the evidence to back-up the correct answer to the question.
295: Opportunity for further education for readers – perhaps include how long the treatment can be provided beyond the 4.5 hours or what instances allow for increased treatment.
331: Unequal number of women/men in the study, also what was the breakdown of women vs men as far as roles? I appreciate that it is noted that there might be other factors, but those may need to be explored in greater detail or at least identified.
Additional comments: 91 (and throughout): Remove “we”. Change to something like “Therefore, the researchers' goal for the study was to determine…” or “The researchers aimed”
100: Similar to the previous comment: “A convenience sample followed by snowballing was used…”
370: While it is unsurprising that the individuals with the lowest scores see the fewest patients with stroke, the question is what does this mean long-term or what does this mean for Malaysia? If a facility rarely sees individuals with stroke and does not have access to advanced care, is the goal for those practitioners to be able to identify symptoms and make appropriate referrals? Or is the goal to increase facilities with advanced care available? Education is still the key component in either scenario.


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: KNOWLEDGE OF ACUTE STROKE MANAGEMENT AND THE PREDICTORS AMONG MALAYSIAN HEALTHCARE PROFESSIONALS
Review round: 2
Reviewer: 1

Output:
Basic reporting: English grammar: the article needs to be corrected so that its of high quality. If you have any programme like grammarly, it will assist you.

Other sections are satisfactory, but more comments on additional comments category below
Experimental design: Satisfactory
Validity of the findings: Satisfactory
Additional comments: Review comments
Thank you for addressing and clarifying my queries/ questions, the manuscript is now taking shape and getting better. Despite that, there are some parts that need further clarification.

More review comments
Data measurements
1.Section 2: ASMaQ. This is the first time it is mentioned, therefore should be written in full with the abbreviation in brackets.
2. Outcome variables
Grammar: Outcome scores were categorized into … not as

Results
3. Level of knowledge on stroke among healthcare professionals
The highest proportion of good knowledge was noted on GSK (88.5%), followed by ASM (61.2%) and HSM (58.1%) (Table 2).
Was this significant, if so where was the significance between GSK and ASM, or GSK and HSM?
For overall knowledge score, you mentioned the mean score with confidence interval. Would be best to mention the other mean scores with confidence intervals too.

4.  Paragraph 2 (208-212): Rephrasing the sentence will make it easier to read and understand e.g. For GSK among HCPs, the highest proportion of poor responses (undesired and neutral response) was noted on item GSK-10 [Acute stroke management education should be conducted regularly for healthcare professionals] (99.9%)…..
The same applies to paragraph 3 (214-217): For HSM, poor knowledge was noted on item HSM-8 [All acute stroke patients must have a 12 leads ECG before thrombolysis] (93.1%),….
Also paragraph 3 (219-223), paragraph 4 (229-231), .
Mention also confidence intervals of the mean scores unless if your text/wording exceed the requirements

5. GSK
Paragraph 1 (252-253): Rephrasing/rearranging of words to make the point visible and easily understandable. Also, include p-value.

6. ASM
Paragraph 1 (264-266): repetition of ASM (at the start of the sentence and at the end). Rearrange the sentence e.g., For ASM, significant factors for poor knowledge were ….
Also, include p-value.

7. It is best to report results using past tense, not mixing past and present tenses

Discussion
8. Some of the texts in brackets are in italics while some are not, is there any reason? Lines 333, 343, 359, 364 etc.

9. Thrombectomy should be performed after thrombolysis in large vessel occlusion in AIS patients who arrive within 6 hours……
Can thrombectomy not be given without thrombolysis?

10. Line 400: should be module not modules

11. Line 371-373: It needs rephrasing to make the point clearer and easy understandable e.g., For ASM, predictors of poor knowledge were short duration of services, not seeing stroke patients in daily practices, working without specialists, and unavailability or being unaware of thrombectomy services in the workplace

Other
12. Missing subsections before references: Information on funding, competing interests, availability of data and materials, and author contributions

Thank you in advance


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 FECAL MICROBIOTA OF THAI SCHOOL-AGED CHILDREN ASSOCIATED WITH DEMOGRAPHIC FACTORS AND DIET
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper started off nicely, with an abstract and introduction that clearly stated the background and aim of the study. All of the supplementary files are provided, but there are far too many files and some files are unclear.
The result part is overly explained, with all of the statistics specifications used are mentioned. It is highly encouraged for the authors to do a major revision on regards to the result part. A more thorough comment on which result part should be revised can be seen in the attached document of this review.
Experimental design: It is easy to find the research question and the research question itself is well-defined and relevant. The experimental design is explained well and replicable. However, it is not necessary to re-mention the statistical analysis in the result part. If it is needed, the statistical analysis method can be added as a footnote in the figure or table to simplify the result part.
Validity of the findings: The provided data are robust and statistically well-defined. But it is quite hard to review as it mentions all statistical methods used that is better to mention them only once in the materials and methods part of the paper. The discussion part is missing some explanation for the results.
Conclusions answered the aim of the study and were well-stated. However, it is unnecessary to re-mention the analysis used.
Additional comments: The importance of the study is well-received by the reviewer and the result will be impactful for future studies. However, there are major revisions needed for this paper including missing parts, structural revision, and overly-explained statistic methods. The author should focus on the results that are very crucial and answer the research question. It is not necessary to explain all of the results obtained. In the end, the reviewer would like to appreciate the authors' hard work and encourage the authors in revising the manuscript.


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 FECAL MICROBIOTA OF THAI SCHOOL-AGED CHILDREN ASSOCIATED WITH DEMOGRAPHIC FACTORS AND DIET
Review round: 1
Reviewer: 2

Output:
Basic reporting: Submitted manuscript describes an analysis of fecal microbiota in a large cohort of children, and their associations with biometric and life-style parameters. An impressive amount of data was collected and processed. Strengthes of the manuscript include large experimentally obtained dataset, a large number of supplementary materials and figures, and inclusion of data. The analytical output is generally well described. However, I have very significant concerns regarding the validity of the data and the choice of multivariate methods. Specific issues are mentioned below:
Experimental design: More information is needed on qPCR raw data analysis and experimental design - how were raw values adjusted to account for unequal amount of starting gDNA copies? Equal amount of total gDNA added into each well does not equal the same number of starting copies as DNA integrity will not be identical among different samples.

The chosen primers have a nested design – e.g., Ruminococcus is in phylum Firmicutes, Prevotella is in Bacteroidetes. While this can be used if one analyzes results for each individual primer set separately, the values cannot be combined in multivariate analyses because they are not independent. I.e., values for Fimicutes will include Ruminococcus abundance, and so forth. Thus, when running PSL-DA analyses, for example, please use variables at only one taxonomic level, e.g., genus.

Why many demographic variables are arbitrarily (e.g., age < 8.05 years, why not 8.00 or 8.11 or …) split into groups? Why not use the full range of values and do a regression-type analysis which had better statistical power?
Please explain why MFA was chosen as the main multivariate analysis tool, rather than other, in my opinion, more appropriate methods such as RDA or CDA? What are MFA advantages that make it the ebst choice for your particular dataset structure?
Validity of the findings: The qPCR data is pretty unreliable: for example, from Figure S12 one can see that total amount of Prevotella (genus within phylum Bacteroidetes) is higher than actual total Bacteroidetes (the same is true for genera of Firmicutes). Were the actual amplification rates incorporated into the CT calculations? Which of the values should the reader trust? Because both cannot be true simultaneously.

Lines 2221-222: “Bacteroides was highly correlated with dimension 1” – indicates that association is obtained post-hoc. Lines 246-247: “In component 2, Lactobacillus and Ruminococcus were the most discriminative bacteria in HFF-low risk”  - indicates that this is obtained during discriminant analysis and is part of the discrimination model. So, which is the correct description of what was done?
Additional comments: MFA figures contain circles that likely represent confidence clouds. This is, however, not described in the figure legends.
There is no taxon called Bifidobacteria, genus name is Bifidobacterium
Some variables are not well described – e.g., the cohort includes school-age children but there is a variable describing formula-fed vs breast-fed children. I assume this represents the feeding mode in infancy, not at the time samples were collected. Please state this clearly.
Many bee swarp plots do not have any explanation for the values (i.e., units)  shown on the Y axis (e.g. Figure 2CDE)
Please use the same test for the same type of data – e.g., panel 2D shows Anova results, but 2E – Wlech test.


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 FECAL MICROBIOTA OF THAI SCHOOL-AGED CHILDREN ASSOCIATED WITH DEMOGRAPHIC FACTORS AND DIET
Review round: 2
Reviewer: 1

Output:
Basic reporting: The reviewer would like to thank the authors for taking our suggestion in revising the manuscript. Most of all the comments has been thoroughly accepted and changes has been made. However, there are still some points that are need to be revised. First of all, some of the subheadings are still not yet corrected (refer to the document attached in this review). Then, limitations part(preferably after the discussion part) can be added to explain the lack of sample size calculation and that cultural practices data cannot be collected due to language barrier.
Experimental design: As previously mentioned, the sample size calculation was missing from the original manuscript. The author replied that the recruitment was based on voluntary participation; hence, no calculation was made. The magnitude base of mean difference was investigated using multivariate statistical analysis. Although it may be clear that this method was done for some, it would be best to provide supporting reference that also applied this method as well.
Validity of the findings: No comment
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: SERUM EXPRESSION OF VASCULAR ENDOTHELIAL-CADHERIN, CD44, HUMAN HIGH MOBILITY GROUP B1, KALLIKREIN 6 PROTEINS IN DIFFERENT STAGES OF LARYNGEAL INTRAEPITHELIAL LESIONS AND EARLY GLOTTIS CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: The rationale for choosing the markers is not sufficiently described in the manuscript. The introdcution should include more about the markers, and their function, along with a brief, but clear background on why the authors chose these markers.

Throughout the manuscript, there are sentences that are not clear and should be re-worded to ensure that an international audience can clearly understand the text. Some examples include lines 58-59, 78-80.
Experimental design: The findings of the study can be made comprehensive by performing in-silico analysis on transcriptomic data to determine the importance of the specific markers in LSCC vs normal tissue.
Validity of the findings: No comment
Additional comments: The research study by Żurek et al describes the validation of molecular markers- Vascular Endothelial-Cadherin Complex (VE-cad), CD44, Human High mobility group protein B1(HMGB1), and Kallikrein 6 in serum samples from patients with specific stages of laryngeal intraepithelial lesions to serve as diagnostic tools in differentiation of benign and dysplastic lesions from invasive pathologies. The research study is novel, and the authors provide a strong rationale for carrying out the study to identify novel biomarkers preoperatively in serum or saliva to guide therapeutic decisions in laryngeal squamous cell carcinoma. The authors have comprehensively discussed the pitfalls and shortcomings of the study and methods. The authors found the markers under study to not be reliable biomarkers implicating malignant potential within vocal fold hypertrophic intraepithelial lesions.


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: SERUM EXPRESSION OF VASCULAR ENDOTHELIAL-CADHERIN, CD44, HUMAN HIGH MOBILITY GROUP B1, KALLIKREIN 6 PROTEINS IN DIFFERENT STAGES OF LARYNGEAL INTRAEPITHELIAL LESIONS AND EARLY GLOTTIS CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript entitled “Serum expression of Vascular Endothelial-Cadherin, CD44, Human High mobility group B1, Kallikrein 6 proteins in different stages of laryngeal intraepithelial lesions and early glottis cancer” by Michał Żurek et al. investigated the potential value of VE-cad, HMGB1, CD44 and KLK6 in serum samples from patients with non-dysplastic lesions, low-grade dysplasia, high-grade dysplasia and invasive cancers as diagnostic markers. They have found that the four proteins are not associated with the stage of laryngeal lesions.
Experimental design: VE-cad, HMGB1, CD44 and KLK6 were not related to disease stages in the human cohort in this manuscript. Authors could devise a diagnostic signature including these four proteins and measure whether the expression of the 4-gene signature is correlated to the stage of laryngeal lesions.
Validity of the findings: Are the four markers, VE-cad, HMGB1, CD44 and KLK6 associated with survival and disease stage of patients with laryngeal cancers? Are they differentially expressed in tumor compared to normal tissue? To validate the findings in this manuscript, authors could investigate in published human cohorts, like TCGA and METABRIC.
Additional comments: None.


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: SERUM EXPRESSION OF VASCULAR ENDOTHELIAL-CADHERIN, CD44, HUMAN HIGH MOBILITY GROUP B1, KALLIKREIN 6 PROTEINS IN DIFFERENT STAGES OF LARYNGEAL INTRAEPITHELIAL LESIONS AND EARLY GLOTTIS CANCER
Review round: 1
Reviewer: 3

Output:
Basic reporting: In the discussion part, for each biomarker, the author compare their expression levels between different cancers, which is not very scientific.
Experimental design: 1.	No legend for all figure and tables
2.	Please label p value on figure1.
Validity of the findings: 3.	Please point out the clinical meaning of this study, which was not spelled out very clearly.
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: SERUM EXPRESSION OF VASCULAR ENDOTHELIAL-CADHERIN, CD44, HUMAN HIGH MOBILITY GROUP B1, KALLIKREIN 6 PROTEINS IN DIFFERENT STAGES OF LARYNGEAL INTRAEPITHELIAL LESIONS AND EARLY GLOTTIS CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: Authors have successfully responded to my comments.
Experimental design: None.
Validity of the findings: None.
Additional comments: None.


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: SERUM EXPRESSION OF VASCULAR ENDOTHELIAL-CADHERIN, CD44, HUMAN HIGH MOBILITY GROUP B1, KALLIKREIN 6 PROTEINS IN DIFFERENT STAGES OF LARYNGEAL INTRAEPITHELIAL LESIONS AND EARLY GLOTTIS CANCER
Review round: 2
Reviewer: 2

Output:
Basic reporting: Thanks for the changes made by the authors point by point.
Experimental design: Good.
Validity of the findings: Good
Additional comments: No.


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: MICROPLASTIC INGESTION BY COMMERCIAL MARINE FISH FROM THE SEAWATER OF NORTHWEST PENINSULAR MALAYSIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article "Microplastic ingestion by commercial marine fish from the seawater of Northwest Peninsular Malaysia" written by Foo, Zanuri et. Al, shows an interest in microplastic consumption by four different commercial fishes in Malaysia.

Microplastic ingestion in marine organisms is well documented around the world but new results are always welcome since this type of pollution is still increasing and solutions for the problem are far from becoming a worldwide reality.

Although the paper abstract and introduction are relevant and well documented as well as figures and data, the quality of the English language must be improved. Among several English difficulties I could point out orthographic problems (such as “tophic” instead of trophic at line 107), sentence repetition (lines 347-349 and 350 – 352) and long and overloaded phrases. This manuscript should be reviewed for its English prior to publication.

The manuscript structure complies with the journal norms and the raw data was supplied.
Experimental design: The research fills the aim and scope of the journal, since it tries to understand the microplastic ingestion by commercial fish in Malaysia. The authors aimed to elucidate the level of microplastic contamination in fishes, which was explained very well along the manuscript. It obviously fills an important gap in the Asian microplastic knowledge since literature lacks information on sea and marine organisms’ contamination from the area.

The experiment was designed with no negative control. This can be understood since there is a lack of microplastic pollution dispersion information in the region making it impossible to the authors to compare the commercial fishes microplastic ingestion to fishes’ contamination from cleaner regions.

The number of individuals analyzed was good, 72 in total, which would be enough to perform a robust statistical analysis which was not shown. This could improve results since the authors would be able to compare the number of microplastics ingested from the two different fishing areas. These results would allow authors not only hypothesize about different microplastic pollution degrees in the two different sites but to have evidences that this may be occurring.

The fish manipulation and digestion protocols are well documented allowing others to replicate the experiment with other species and areas so results could be compared.

Work with microplastics identification should be done with rigorous protocols to avoid external contaminations. Authors indicates they were aware of the problem, but even though precautions were made, the control digestion experiment detected some microplastic presence, which could be something to explain better, or to insert in their data analysis.
Validity of the findings: Results are well described, and the raw data was attached to eventual observations. The total number of individuals is good enough to perform statistical analysis which I was not able to identify in the manuscript. The figures are well labeled but in the manuscript the authors go from figure 5 (line 278) to figure 8 (line 289). It seems to me that figure 6 and 7 are not referenced in the text thus it shows the microplastic particles on microscopical views which is interesting since until there, there was no mention of microplastic observation.

The SEM-EDX results could be improved and references should be annotated (from line 313 to 318 the authors affirm that presence of carbon and oxygen, potassium and calcium are indication of plastic but in any moment they indicates a literature to substantiate findings).

The conclusions are well stated and limited to the findings. Humans' microplastic contamination possibility and effects are speculated and identified as something remote since there are not enough studies to confirm it, even if microplastic may be found in consumable fish parts.
Additional comments: The manuscript is interesting and well designed but it lacks english lanquage quality that should be assessed before publication, The results have potential to initiate other experiments and new insights about plastic contamination in the area of study which is always something good to the future of your research.
The difficulties of analyzing environemental sampled microplastics are inherent to the ubiquity of ther presence, but your work, using different analysis methods was able to not only identify microplastics but to try to indicate what type of plastic there were in the gut of commercial fishes.
I guess that with robust statistical analysis and a better english language, the article wil be more pleasant to read.


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: MICROPLASTIC INGESTION BY COMMERCIAL MARINE FISH FROM THE SEAWATER OF NORTHWEST PENINSULAR MALAYSIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: - Language and grammar issues: The English language needs be improved to ensure that an international audience can clearly understand your text. Some examples where the language could be improved along the introduction– the current phrasing makes comprehension difficult. I suggest you have a colleague who is proficient in English and familiar with the subject matter review your manuscript, or contact a professional editing service.

- Introduction, background & discussion: Can still be improved and more aligned

- Literature was relatively well referenced & relevant ☺

- Figures are relevant and images are of good quality. Labeling & description can be improved.

- Raw data was supplied

 - No hypothesis was tested or clear research question attempted to be answered. It is important to expand upon the knowledge gap being filled. To me it seems that the major knowledge gap filled was in regards to contributing to data collected in Malaysia.
Experimental design: - Sampling design was intrinsically confounded by species analysed in the different sampling location/ dates.  Crenimugil seheli (collected on November 27th, 2019 at the
location 1) was compared to Atule mate, Sardinella
fimbriata and Rastrelliger brachysoma (collected on
December 3rd, 2019, on location b). You nicely discussed the possible issues related to differences in the sampling dates, locations and other factors such as feeding behaviour (lines 405-414 and 433-466).

- Methods were described with sufficient detail & information to be replicated

- Results on the average abundance (numbers in each species and numbers in individual fish, figures 3 and 4) indicated that amounts of plastics were relatively similar among the three species collected from location 2, although slightly greater to Sardinella. It seems that if you run a statistical test (e.g. analyses of variance), you would find no significant differences in the abundance of plastics among the three sps collected from location 2. After having results from a statistical test, the emphasis you gave Sardinella having the highest abundance should be reviewed on results, discussion and conclusion.
Validity of the findings: The data provided is a valid and adds information about the presence,  abundance and types of micro plastics found in the guts of commercial fish .

However, discussion and conclusions were not clearly connected to a research question and conclusions were not limited to those supported by results from this MS.
Additional comments: General comments:
There is some interesting information in the MS, albeit not unexpected, but it needs important conceptual change and revision to make it a useful contribution to our understanding of the microplastics ingestion by pelagic fish more generally and to its possible transference to humans.
I have made some comments using sticky notes in the PDF and have elaborated some key points below.

Detailed comments

Abstract
Line 62: Replace ‘fishes’ by ‘fish’. The plural of fish is actually fish … awkward things of the English language. Check all the remaining text…

Introduction

Remarks:
Were there any hypothesis being tested?

The significance of the work needs to be more clearly stated during the introduction. Although microplastics ingestion by fish have been previously reported around the world, there is relatively small information from Malaysian’s commercial coastal species? Are there any other significant contributions? Explain it please

Lines 75-78: Please revise the concepts of degradation and biodegradation and the link between ideas was not clear.
I suggest rephrasing as: Plastics are extremely resistant to biodegradation. They, however, degrade into smaller particles over time when predisposed to several natural factors, such as sunlight and wave action (Wang et al., 2016).
Delete: the bit “and therefore rapidly accumulate in marine ecosystems that could be very
harmful especially for the marine environment”. (Does not link with previous idea and, its not because they degrade over time that they accumulate in marine ecosystems…

Lines 81-82: The increase reach of plastics in natural environments is also due to its mismanagement. I suggest rephrase it as:
“Plastics will
 increasingly reach all areas of our environment inherent to the growth of plastic production (Azoulay et al., 2019) and its mismanagement during production, distribution, use and final disposal.

Line 91: Check definition of primary microplastics. Not all of them are produced as pellets.  Primary microplastics are I intentionally produced by industries within this size range, for direct applications (i.e., medicine and cosmetics). Primary microplastics include industrial ‘scrubbers’ used to blast clean surfaces, plastic powders used in moulding, micro-beads in cosmetic formulation, and plastic nanoparticles used in a variety of industrial pro- cesses. In addition, spherical or cylindrical virgin resin pellets…  (see “GESAMP (2015). “Sources, fate and effects of microplastics in the marine environment: a global assessment” (Kershaw, P. J., ed.). IMO/FAO/UNESCO-IOC/UNIDO/WMO/IAEA/UN/UNEP/UNDP Joint Group of Experts on the Scientific Aspects of Marine Environmental Protection). Rep. Stud. GESAMP No. 90, 96 p.
Line 99:  Primary microplastics can also be ingested. Suggest rephrase as: ‘Primary and secondary microplastics have the potential to be ingested …’
Lines 115 – 128: that hole part belongs to the methods

Materials and Methods

Remarks: It was not clear if you used an already available standardized protocol OR if you a proposing a new protocol. From your text and references it seems that you’ve used a combination of previous protocols from Lusher et al., 2017 and Dehaut et al., 2016.
 If that’s correct, why did you adapt protocols?

Lines 136-140: Your sampling design is intrinsically confounded by species analysed in the different sampling dates. It’s not clear why you collected Crenimugil seheli
on the first sampling date and other three different species on the second…This rational or problems associated with your sampling strategy need to be explained.

Results

Remarks:
Do you have data about the actual sizes of microplastics found in your samples? This would greatly contribute to a possible speculation about transference to humans via consumption of fish

Review your statements about greatest abundance of microplastics in Sardinella

Lines 272 -277: Needs to be re-structured / rephrased
Suggestion would be to say  “Meanwhile, the abundance of micropalstics was greater on three species collected at location 2. Relatively similar abundances were among the three species collected from location 2, although slightly greater to Sardinella.”

Discussion

Line 347: Start a new paragraph

Lines 350-351: Delete, repetition of previous phrase.

Lines 364-367: Concentrate your discussion around your research focus, ingestion by fish under natural conditions.

Lines 367-370: “The result of
 this study coincides with the study of Browne et al. (2010) as well, which stated that the
arising risks of microplastic ingestion by marine animals indicates microplastic is more
abundant than large plastic debris in the marine environment now.” Suggest to remove that bit or rephrase it …. Browne et al. (2010) evaluated spatial patterns of macro and microplastics fragments. Their results showed that microplastic accounted for 65% of debris recorded. They mentioned that when macroplastics fragments into smaller pieces, the potential for ingestion and accumulation within the tissues of animals increases. They don’t stated that arising risks of microplastic ingestion by marine animals indicates microplastic is more
abundant than large plastic debris in the marine environment now…
Lines 419-420: “Sardinella fimbriata is
planktonic organisms and filter feeders”. That needs to be rephrased… Did you mean to say “Sardinella fimbriata are filter feeders and feed on planktonic organisms?
Lines 478 – 503: Possible impacts on human’s health can be related to the amounts and sizes of microplastics ingested, among other factors. Having this in mind, if you are going to speculate about transference to humans, it would be important to explain if guts of the four species are normally ingested as human’s food or if only the fish filed is consumed. Also, if possible please provide information about the sizes of the microplastics found on your samples and state the sizes of MP found by other authors (e.g. Abbasi et al., 2018)

Conclusions:

Remarks: needs to be linked to original research question & limited to supporting results.
See detailed comments in the text.


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: MICROPLASTIC INGESTION BY COMMERCIAL MARINE FISH FROM THE SEAWATER OF NORTHWEST PENINSULAR MALAYSIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: changes were made accordingly to reviewers suggestions
Experimental design: changes were made accordingly to reviewers suggestions
Validity of the findings: changes were made accordingly to reviewers suggestions
Additional comments: To explain the tested hypothesis, authors made a bullet list in Material and Methods. It would be better to incorporate the hypothesis at the end of the introduction as a discursive text.


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: MICROPLASTIC INGESTION BY COMMERCIAL MARINE FISH FROM THE SEAWATER OF NORTHWEST PENINSULAR MALAYSIA
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors revised the text according to prior reviews.
Experimental design: The authors revised the text according to prior reviews.
Validity of the findings: The authors revised the text according to prior reviews.
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: UNLOCKING ANDEAN SIGMODONTINE DIVERSITY: FIVE NEW SPECIES OF CHILOMYS (RODENTIA: CRICETIDAE) FROM THE MONTANE FORESTS OF ECUADOR
Review round: 1
Reviewer: 1

Output:
Basic reporting: This study is written very well.  The authors went to great efforts to assure that it is grammatically correct.  It is easily understandable by all readers.  The introduction is thorough; its details welcome. The procedures and methods of the study are laid out nicely.  Figures are clear, tables are thorough, references are complete.  Correct the tittle of Appendix 1 ('studied' specimens). This study is self-explanatory and self-explained.
Experimental design: I like this paper.  It lays out the research objective clearly and reports on relevant information, in this case demonstrating diversity within Chilomys leading to diagnosis of new species,  The standards of research are quite high and rigorous.  Relevant ethical and scientific guidelines are acknowledged. Methods are eminently reproducible and testable.
Validity of the findings: This is a highly original study, completely appropriate and an advance for the field of research.  The results are a useful, positive contribution.  The conclusions flow from the observations and are well-summarized; future work is briefly outlined. Relevant data are presented in detail in the main article and in supplementary files.
Additional comments: This paper is done well and I appreciate the efforts of the authors to assure readability for an international audience.  I have marked the pdf with edits that I hope are helpful and do not change meaning.  Some are meant to help the narrative, sometimes by shortening it.  Authors should check the preferred spelling of 'procumbency' and use it consistently.


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: UNLOCKING ANDEAN SIGMODONTINE DIVERSITY: FIVE NEW SPECIES OF CHILOMYS (RODENTIA: CRICETIDAE) FROM THE MONTANE FORESTS OF ECUADOR
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is clear and technically correct. In general, the article includes a sufficient background about the taxonomy of the group under study. However, I consider that it is necessary to emphasize the phylogenetic relationships of the genus with other genera of Sigmodontinae rodents, as that would help to understand the sampling of external groups in phylogenetic analyses. Furthermore, the Introduction should specify why this study is restricted almost exclusively to Ecuador, even though there are populations in Venezuela, Colombia, and Peru. In this sense, I think that the information indicated in the first paragraph of the results should be raised in some way in the introduction. Thus, it would be possible to know the scope and limitations of this taxonomic study.  I also recommend following a more logical order in the presentation of evidence throughout the entire manuscript. If the authors decide to start the materials and methods with the morphological part, then I think that this should be presented at the beginning of the results (and not the molecular results).
Experimental design: Morphology
•	It is not completely clear what is the purpose of the X-ray Micro CT. Is it used as a source of evidence to distinguish potentially new species? Or, once the lineages were detected, is it used to better characterize those new species? In addition, it should be explained why this process was performed only for three specimens (holotypes). On the other hand, the author uses two multivariate analyses (PCA and DFA), which are very common in taxonomic studies. However, sometimes they are used without solid justification. I highly recommend specifying what the objective of each of the two analyzes is and thus justify their use. See, for example, Strauss, R. E. (2010). (pp. 73-91). Springer, Berlin, Heidelberg.

Genetics
•	It is not clear how many specimens were sequenced. Following up on my comments about multivariate analyses, Cytb and COI are commonly used in mammalian taxonomic studies. I think the authors should at least briefly mention why they use both and not just one, considering they are mitochondrial. I also recommend specifying the PCR protocols used for this study, especially if the authors are using a combination of two protocols (i.e. Bonvicino & Moreira, 2001; Smith & Patton, 1999).
•	The phylogenetic analyses should be better explained.  Hoy many sequences were included in the analyses? What was the length of the amplified fragments per gene and what was the result of the assembly? Is there missing data? Were the same analyses done for each gene separately and also for the two genes put together? Looking at the supplemental material, I see the author used Rhagomys, Thomasomys, and Rhipidomys as external groups, but that should be clearly mentioned and justified in the text. In the discussion, the authors mention that Aepeomys is sister to Chilomys; then, why this genus was not included as an external group?
•	The authors mention that they also used sequences from GenBank. Please, specify what other sequences you mean. Other sequences of the genus Chilomys? Sequences of external groups? Sequences from other studies?
•	It should be specified whether the genetic distance analysis was done for each gene separately or both concatenated.
Validity of the findings: •	The author mentions that Chilomys is “embedded in a clade with Rhipidomys, and Thomasomys”. Although this is true, I think this finding is obvious since they were the only genera used as outgroups ((and rooted with Rhagomys). The Materials and Methods should contain the rationale for why these genera were selected as outgroups. Because the results for the Cytb and COI genes vary as to the relationship of Chilomys to the Rhipidomys and Thomasomys, I think the authors could extend the outgroup sampling to discuss the monophyly of Chilomys and its sister group. If it is not possible to expand this sampling to carry out new analyses, then I recommend that the authors expand the discussion around the possible uncertainty of the relationship of Chilomys with other genera.
•	Regarding the analysis of genetic distances, I think the authors only present the result for the Cytb (Table 1). Why is the COI result not presented?
•	In my opinion, the results from the two multivariate analyses are poorly explained and underutilized. How is the variation distributed among the samples? What are the variables that most contribute to this variation? Are there variables that help discriminate some species more from others? Why is a classification analysis not shown for the samples?
•	Perhaps my biggest comment is regarding the decision to consider 5 new species. It is not clear how the authors integrate the results from the phylogenetic analyses, the species delimitation methods, and the morphometric analyses. For example, the Cytb phylogeny seems to support five lineages, but the COI maybe three or four. The delimitation method even suggests many more than five species. The morphometric study is uncertain since the authors seem to use the five groups identified by the phylogenetic analysis of Cytb as a priori lineages. I recommend adding a section in the results section that integrates the evidence and thus justifies the recognition of 5 species. Overall, I suspect that the authors decide to base their conclusions on the phylogenetic tree derived from the Cytb sequences. I am not saying that it is a wrong decision, but that it must be clear and well justified based on all the evidence that the authors present.
•	Given the sampling (practically restricted to Ecuador), I highly recommend that the authors mention the limitations in the discovery and description of these 5 species for Ecuador. In addition, what are the limitations of the genus diagnosis if samples from other countries are not reviewed in detail?
•	The discussion about the diversification of the genus Chilomys is very interesting, but I think it is outside the scope of this study and appears to be speculative. Therefore, I suggest that that part be removed from the present manuscript.
Additional comments: I suggest that the list of examined specimens be treated as Appendix and not as supplementary material. I think the specimens examined are the most important part of taxonomic reviews and should not be relegated to supplementary material.


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: UNLOCKING ANDEAN SIGMODONTINE DIVERSITY: FIVE NEW SPECIES OF CHILOMYS (RODENTIA: CRICETIDAE) FROM THE MONTANE FORESTS OF ECUADOR
Review round: 1
Reviewer: 3

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: I am sending to you my suggestions and appreciation of the manuscript entitled “Unlocking Andean sigmodontine diversity: Five new species of Chilomys (Rodentia: Cricetidae) from the montane forests of Ecuador” (#68489).

The work done by Dr. Brito and his collaborators is clear, complete and follows the rules and norms of the journal. Authors followed the criteria and the high quality of the figures and tables, and citations and literature are impeccable. The manuscript is clearly written in professional and unambiguous language.
I consider the authors made excellent fieldwork efforts, and after many years of research in scientific collections, they compiled an extensive dataset that includes external and internal morphological data of the different species here proposed, including information obtained from x-rays of the skull, which makes this work a reference for the proposal of new mammal species.
However, the molecular analyses are not as complete as the morphological ones (it was not included any nuclear marker), but all other evidence is trustworthy, including the support of the different mitochondrial clades.
I only disagree in the recognition of the clade Chilomys carapazi sp. Nov. Brito and Pardiñas as an independent species, and I would recommend discarding it as a new species for the following reasons. After analyzing the diagnosis of the species and the information obtained throughout the analysis of this single specimen, I found the proposal very risky, since there is no molecular evidence that supports it, and the morphological analysis was based is an old specimen with a wearing teeth pattern that does not correspond to the ages of the rest of the individuals (Fig. 8). In other words, even though the PCA analysis showed this specimen was different, the comparison with other specimens is not clear (and possibly not valid because authors could be just comparing specimens of different ages). The lack of strong differentiation of that specimen resulted in its recognition as new species based on the combination of multiples characters, which in my opinion is not enough to recognized it as a different species.


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: ASSOCIATION OF HEAT SHOCK PROTEIN 8 WITH ATOPIC MARCH IN A MURINE EXPERIMENTAL MODEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this study Kyu-Tae et al., propose an improved murine model for atopic march and suggest HSP8 as a candidate biomarker. Authors have applied low-dose of allergen and hence have minimized skin irritation when inducing atopic dermatitis. Using this improved methodology they have assayed IgE level, Th2 response cytokine and lastly have done proteomic analyses to find candidate biomarkers.
The experimental design is methodical and understandable. Manuscript is well-written.
Experimental design: Methodical and well designed
Validity of the findings: Conclusions are well stated and authors have discussed limitations of the study
Additional comments: Authors have done a very good job of writing this manuscript in clear and unambiguous language which is very important to understand the goal of the study. Data provided in the manuscript is also convincing and clean. I also commend authors for discussing the limitations of this study. I have few other comments about this manuscript:
1.	In ELISA assays (Fig4), authors did not observe differences in IFN-gamma. To establish that Th response is more of Th2 type and not Th1 type, they can also do ELISA for IL-2 and IL-12, cytokines hallmark of Th1 response.
2.	Authors have suggested HSP8 as a candidate biomarker using proteomic data analyses. A simple WB showing expression of HSP8 in the groups compared in the study will further validate this observation. I guess this simple experiment will be under scope of this manuscript.
3.	Assays showing TSLP, IL-33 and IL-25 expressions will further validate this improved model for atopic march.
4.	Is it possible for authors to provide the fold changes for the proteins depicted in fig 5. It may prove helpful to prospective readers.
5.	Lastly, I was just wondering if the list of proteins and (particularly HSP8) the authors have got from proteomics data show modulated expression in response to low-dose of allergen used to induce AD. In future, it will be interesting to see if HSP8 and its interacting partners are still modulated at higher/different doses of allergen. Also, it will be interesting to see if HSP8 and its interacting partners are modulated in other models of atopic march studies. Such studies will help establish HSP8 as a reliable candidate marker for atopic march irrespective of dose or models used for the study. These are just suggestion as a part of peer-review and authors are not needed to respond to this particular comment.