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: GENETIC DIVERSITY AND STRUCTURE OF THE NOBLE CRAYFISH POPULATIONS IN THE BALKAN PENINSULA REVEALED BY MITOCHONDRIAL AND MICROSATELLITE DNA MARKERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: -The article is written in clear, unambiguous, technically correct English text.
A few points that need attention:
L265. Α comma after ‘(655 bp long)’ is needed.
L446. ‘Study also revealed’. The word ‘also’ might be removed.
L779, L913. Journal in italics.
L983-984. ‘Add your references here’ has to be removed. Please check the references’ formatting one more time.

-The article includes sufficient introduction and background information and the study fits well into the broader field of knowledge for the species.

-The article is well structured and all Figures and Tables are relevant to its content with sufficient resolution, appropriately described and labelled. Also, all appropriate raw data have been made available in GenBank.
A few points that need attention:
FIGURE 1. In the down-right corner of the figure, it is stated the Black Sea. This is actually part of Greece and it is the Aegean Sea. The Black Sea is not visible in this figure.
TABLE 1. For site Prespa, the coordinates indicate L. Prespa as the sampling site but it is stated L. Ochrid as the river basin and major tributary. It is not very clear.
L292-297. Are you referring to Fig. 2A, particularly?
L301. Please correct ‘3)’ to ‘(Fig. 3)’.

-The article is self-contained, representing a unit of publication, and includes all results relevant to the hypothesis.
Experimental design: -The article contains clearly defined and meaningful research questions. The knowledge gap being investigated is also stated clearly and statements are made as to how the study contributes to filling that gap.

-The investigation has been conducted rigorously and to a high technical standard. The research has been conducted in conformity with the prevailing ethical standards in the field.
Just a point that needs attention:
The number of samples per population varies from 9 to 33 (Table 1). Nine individuals are quite a few to statistically represent a population. How did you confront this issue? Please mention it also in the Discussion.

-The Materials and Methods of the study are described with sufficient information, in general, to be reproducible by another investigator.
A few points that need attention:
L208. ΦST is presented (also in L345, L346 L556) instead of the FST fixation index. Please explain.
L227-228. Please provide details on how the microsatellite genotypes were scored. Genotyping is a very bias-sensitive procedure. Have they been scored automatically? Were they manually double-checked and from how many experts? Is there any sample, or more, double analysed to ensure the genotyping consistency? What else did you do to avoid any relevant bias?
Validity of the findings: -Decisions are not made based on any subjective determination of impact, degree of advance, novelty or being of interest to only a niche audience. We will also consider studies with null findings. Replication studies will be considered provided the rationale for the replication, and how it adds value to the literature, is clearly described. Please note that studies that are redundant or derivative of existing work will not be considered.

-The vast majority of data on which the conclusions are based is provided or made available in an acceptable discipline-specific repository. The data seems robust, statistically sound, and controlled.
Just a point that needs attention:
L363-364. Why are data not shown? There are other 3 instances in which data are not shown (L350, L367 and L405). Could you please explain each one?

-The conclusions are appropriately stated and connected to the original questions investigated, limited to those supported by the results.
Additional comments: I have read thoroughly the article ‘Genetic diversity and structure of the noble crayfish populations in the Balkan Peninsula revealed by mitochondrial and microsatellite DNA markers’ which address the understudied genetic diversity and population divergence of noble crayfish populations from the western part of the Balkans using mitochondrial and microsatellite DNA markers. The study fills a knowledge gap and provides useful data for conservation planning and management of the species populations.
I would like to congratulate the authors for their clearly written manuscript which is based on large data set, compiled over many years of detailed fieldwork (2008-2016). If I must state some points, as I stated above and in order of importance, which should be improved upon before Acceptance, these are:
1.	Clarification on the sample sizes bias of the studied populations
2.	Microsatellites scoring
3.	ΦST vs FST usage
4.	Explanation of why some data are not shown
5.	Figure1 and Table 1 corrections
6.	Just a few linguistic corrections


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: GENETIC DIVERSITY AND STRUCTURE OF THE NOBLE CRAYFISH POPULATIONS IN THE BALKAN PENINSULA REVEALED BY MITOCHONDRIAL AND MICROSATELLITE DNA MARKERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The finding of this research is very interesting. I would like to suggest the editor accept this manuscript with minor revision. The manuscript is well structured and written, however, there are still minor grammatical mistakes that can be observed throughout the manuscript. The methodology used in this research is satisfactory and suitable to be published in this journal. Even though this project is regional, the data presented are sufficient and discussed well, important as a reference for future research due to the translocation of this species to many places of the world for human consumption. I suggest the authors improve:
-	Figure 1: enlarge the country name on the map.
-	Figure 2: make all the characters there readable.
-	Please label the location name of the black dots in Figure 6.


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: GENETIC DIVERSITY AND STRUCTURE OF THE NOBLE CRAYFISH POPULATIONS IN THE BALKAN PENINSULA REVEALED BY MITOCHONDRIAL AND MICROSATELLITE DNA MARKERS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Dear authors,
I was pleased and happy to read through your manuscript bringing light into genetic diversity of native European crayfish species – Astacus astacus. As authors declared, there have been several studies already published, but not considering samples from extant area of southern Europe, especially Balkan countries. That countries have been previously marked as possible genetic diversity refugia for Austropotamobius species and as proven by this study for A. astacus as well. Big effort was done on completing all existing sequences of A. astacus available on GenBank and BOLD databases to present complex structure and diversity of this species. Methods are written clear and detailed. Results are presented precisely with nice figures. Discussion is extensive considering each possible explanation of the results.
I have only very minor comments to the text
Figure 2. The Bayesian phylogram is not well readable due to low resolution. I hope that in final state this will have sufficient resolution. However as this was the case of most of the figures, I guess this is caused due to compression when pdf version of submission was created. Anyway, in the Figure 2, the labels of phylogram are not readable.
Another mistake was found in Supplementary figure 1, where the most of picture is black and all labelling is not visible at all. I don’t know whether this was purpose or a weird artefact, but it needs to be displayed correctly.
Lines 185-186: I would delete bracket from “dataset I) and II)” just dataset I and dataset II are fine.
Lines 423-424: “The lowest Ne values were found in the Romanian population of BEZ (Ne = 11), and the Serbian populations of GRL (Ne = 9) and GAZ (Ne = 18).” – I would switch the two last values being in descending order, i.e., GAZ and GRL.
Lines 495-507: Authors discuss the low genetic diversity of A. astacus comparing to other European crayfish species, however not mentioning the possible relationship to A. balcanicus, the valid species according to Crandall and DeGrave (2017) and their „An updated classification of the freshwater crayfishes (Decapoda: Astacidea)..”. Although, Lake Ohrid is the type locality of this species, adopting sequences from Mrugala et al (2017), the haplotypes were not different enough to show different species status. Most likely also some populations from Laggis et al. (2017) should originated from Vardar river basin, but again didn’t show sufficient divergence. I found the recent paper discussed A. balcanicus status, however dealing with another species A. colchicus (Blaha et al. 2021 Integrative Zoology 2021; 16: 368–378). I would be grateful to authors to add several sentences discussing this issue.


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: POTENTIAL ARABIDOPSIS THALIANA GLUCOSINOLATE GENES IDENTIFIED FROM THE CO-EXPRESSION MODULES USING GRAPH CLUSTERING APPROACH
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall, the reporting is clear. However, there are some sentences that require rephrasing. Here are some specific examples:
-Line 65: “Glucosinolates (GSLs) are nitrogen-containing compounds” the “nitrogen containing” is redundant, please delete.
-Lines 251-252: “We hypothesize that co-exist genes with known GSL genes in the same statistically significant clusters can be used to predict potential GSL genes” – this sentence is not clear. Please rephrase/ explain.
-Lines 328-330: “Sulfur content in GSLs indicates that sulfur in plant is critical in GSL biosynthesis. Several metabolomic and transcriptomic studies reported a significant reduction in GSL accumulation under sulfur deficiency environment, suggesting the role of sulfur in GSL biosynthesis”- redundant, please rephrase.
-Lines 357-358: “In GSL biosynthesis, several groups of GSLs differ from the side chain derived from the amino acids that serve as the precursor” – rephrase.
Experimental design: The experimental design makes sense and is summarized very nicely in Figure 1. However, the reader will benefit if the authors try to engage Figure 1 to the results/methods sections by referring the reader to this figure throughout the manuscript, and by adding the relevant figure references to the steps describes in Figure 1.
Validity of the findings: The findings are well validated. No other comments.
Additional comments: Line 253-254: “Five sets of clusters were generated using density values of 0.5, 0.6, 0.7, 0.8, and 0.9 with cp value of 0.5”- Please elaborate, mainly on the density values and cp value, as part of the results, not only in the methods section, to give your results more context.
-Line 269: “Based on Table 3, the top six highly significant clusters … were chosen” – based on table S2, right? Table 3 is the six clusters.
-Figure 1 is very helpful in facilitating your workflow. Try to engage it to the results/methods sections by referring the reader to this figure throughout the manuscript, and by adding the relevant figure references to the steps describes in Figure 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: POTENTIAL ARABIDOPSIS THALIANA GLUCOSINOLATE GENES IDENTIFIED FROM THE CO-EXPRESSION MODULES USING GRAPH CLUSTERING APPROACH
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors’ provide sufficient field background/context to understand the topic and purpose of the research. Although most of the relevant references are included, the authors’ overlooked an important publication, Wisecaver et al., (2017) The Plant Cell. Wisecaver et al. used an alternative method of gene co-expression analysis to identify specialized metabolite genes, in particular glucosinolate genes, in Arabidopsis thaliana and Brassica rapa. Some potential glucosinolate gene candidates recovered in this manuscript were identified by Wisecaver et al. previously. MRSA2 (AT5G07460) and AT1G21440 were recovered as co-expressed with glucosinolate genes in Wisecaver et al., supporting these genes may be involved in glucosinolate biosynthesis but not as novel candidates. This manuscript should consider referencing Wisecaver et al. and incorporate the findings from that study into the results, discussion, and conclusions of this manuscript.

Although the underlying data used in the study is already publicly accessible through well curated databases, the addition of the original gene co-expression matrix that was generated from these data sources, used in analyses, and referenced on lines 148-150 would further strengthen the quality and robustness of this manuscript.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors' have prepared a well-written manuscript reporting the results of their method of identifying glucosinolate biosynthesis genes and candidates. Identification of specialized metabolite genes through sequence similarity is problematic, and gene co-expression analysis have been shown to successfully recover clusters of genes involved in specialized metabolic pathways. There are many ways of performing gene co-expression analysis and results being dependent on the input gene expression data. This study provides an additional, alternative method of performing gene co-expression analysis to recover glucosinolate genes from the dataset, as well as identify additional glucosinolate gene candidates.

A small factual error stated should be corrected. Line 68 states “GSLs are exclusively found in the Brassicaceae family…”, which isn’t quite correct. Glucosinolates are predominantly produced by Brassicaceae species, but have been identified in other plants outside the Brassicaceae family.


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 EFFECT OF UNCORRECTED AMETROPIA ON OCULAR TORSION INDUCED BY CHANGES IN FIXATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: Introduction: There are several points described in the literature, but no further elaboration to inform the details of the cited studies.
1.	Line 57: “…, but studies on these changes due to eye movement are insufficient compared to other factors.” This statement is not clear. The authors need to explain what are the factors and the short-coming related to these factors. Please include the references.
2.	Line 58: “There are several studies on visual perception and eye movement (Westheimer & McKee, 1975),…” Only one study for this statement. What is the significance of this reference study in relation to your research? This statement needs clarification.
3.	Line 59: “…, but relatively few studies have examined ocular torsion.” Please quote the references for this statement.
4.	Line 60: “The study by …….of the retina.” The authors relate ocular rotation to the peripheral retina. What is the relation of stimulation of the peripheral retina and the relevance to this study's aim?
5.	Line 64: “…, can affect uncorrected ametropia.” Please include references for this statement.
6.	Line 67: “Although there are several methods for measuring ocular torsion…”. For this statement, please list the techniques used in the literature with references.
7.	Line 71: “However, since the ……..fixation distances”. The statement is not clear. The authors are explaining the weakness of the technique. Perhaps, elaborating the amount of change in ocular rotation for a specific range of fixation distance could better clarify this statement.
8.	Line 81: “The method….. iris image (Kothari et al., 2005).” The statement is repeating information in line 67-68. Suggest removing the sentence.
Experimental design: Participants:
1.	Please include age information. Although the age factor may not affect the convergence, it affects the accommodation function. A poorer accommodation function has been related to the instability of fixation at near. It could change the exertion from the vergence system, particularly in certain binocular vision anomalies such as the accommodation insufficiency or accommodation excess. Has the accommodation function been considered in participants’ screening?
2.	Participants who cannot achieve 6/6 vision with optical correction in each eye and binocularly were eligible for the study? For instance, amblyopic patients and patients with high astigmatism (-2 DC) show poor visual quality and likely affect the visual fixation.

Experiment protocols:
1.	Line 109: “The fixation target used point target,…”. When changing fixation to a near object, always near triad happens (miosis, accommodation and convergence) to ensure clear single vision at near. My question, is there any particular reason for using the “point target” as the fixation target? As we know, point target is not a good stimulus for accommodation.
2.	The size of the point target is not informed in the article. Did the authors change the point target size when testing different myopia groups?

Data analysis:
1.	The analysis section is not clear. Suggest the authors elaborate the purpose of the analysis and inform the analysis tools applied. For example, ”To compare the ocular torsion between emmetropes and myopes, unpaired t-test was conducted….”.

Figure 5
Suggest to include “myopic subjects” and number of subject in the title.
Validity of the findings: Participants characteristics:
1.	Please include the number and refractive range of participants with emmetropia, simple myopia, myopia with astigmatism of 1 DC or lesser, and myopia with astigmatism of 1.25 DC or greater.
2.	Also, please include the age range for each of the refractive sub-categories

Results: Line 142 – 191
1.	Overall, the main objective of the paper is to relate the type of ametropia (emmetropia , simple myopia, myopia with low astigmatism, myopia with high astigmatism) and the ocular rotation at 3m and 40cm. Sub-sections in the results are confusing; without reporting which refractive groups were compared, no information of the number of participants included in the type of ametropia group and the simple t-test limit the testing of the interaction effect of within-subject factors (fixation distance and type of ametropia). Perhaps, performing a repeated measure ANOVA (exclude the 3 hyperopes) could simplify the analysis and better present the results.
2.	Based on analysis in point 1, if the type of ametropia did not change the ocular rotation significantly, then, the data can be grouped to test the effect of ocular dominancy and ocular rotation.
3.	Line 148: “It has been reported that……” This statement is inappropriate in the result section. Please move to the discussion.
4.	Line 151 – 152: This statement does not add additional information and redundant to the statement in line 154. Suggest removing the statement.

Discussion:
1.	Line 195 – 197: The results showed the ocular rotation was not statistically significant between emmetropia and myopia. But, the statement reported myopia and astigmatism group showed larger ocular rotation. This statement is inappropriate unless the authors could relate the minimum angle of rotation that impacts the visual functions.
2.	Line 254: “….increased more when ametropia and astigmatism…”. The word “ametropia” is not appropriate since the authors compared the emmetropia and myopia group. Suggest changing it to “myopia”.

Conclusions:
Line 292 – 293: “Therefore, we suggest that a full correction should be performed ……near distance.” I am concern with this statement. Myopia of 3 D or greater is rarely left uncorrected for distance and near vision in real-world practice. What is the significance of this study’s results in relation to clinical practice?
Additional comments: No additional comments


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

Input:
Title: THE EFFECT OF UNCORRECTED AMETROPIA ON OCULAR TORSION INDUCED BY CHANGES IN FIXATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: - One potential confound in the article is the amount of vergence across distances. I imagine this is something that it is not possible to measure at this stage but at least it should be addressed in the discussion as a limitation. It could be that some subjects were more accurately fixating binocularly in the target and thus inducing a different amount of torsion.
- Throughout the authors propose the idea that torsion is somehow used to stabilize the retinal image that moves due to eye movements. I cannot follow the logic of the argument as stated right now in the manuscript.
Additional comments: - Abstract and line 202: It is not correct to say "Ocular torsion, one of the eye movements to stabilize the
retinal image". Ocular torsion is a more generic term simply referring to eye movements when the eye rotates around the line of sight.
- Lines 34-35: These two sentences appear to contradict each other. Appears to say something like, "Eye movement is required .... because eye movements move the retina"?
- Line 35: I would not use the word type I would say components or dimensions.
- Line 42: the sentence "but it can also occur with a static posture depending on the distance of the fixation target" seems a bit out of place. Specially since the next sentence uses the term ocular counter-roll. The next paragraph seems a more appropriate place to introduce torsion due to posture.
- Line 61: Do you mean the studies are smaller (fewer) or that the torsional movements are smaller?
- Line 63: How do torsional eye movement stabilize the image following changes in fixation distance?
- Line 74-83. It is very hard to follow which methods you propose and which ones you actually used?
- Line 109: The sentence seems incomplete "used point target".
- Line 157: I don't understand the logic for using only the dominant eye data. Why not averaging both, for example?
- Line 237: Similar to a previous comment. I don't understand the idea of torsion stabilizing the retinal image that moves due to eye movements.


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: TRANSTIBIAL LIMB LOSS DOES NOT INCREASE METABOLIC COST IN THREE-DIMENSIONAL COMPUTER SIMULATIONS OF HUMAN WALKING
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is well-written in an organized manner and figures are well-presented. However, I suggest that the authors check the references. For example, line 36, “Pinzur et al., 1992” is not found in References. (This was found by chance, and there may be more. Therefore, a thorough check is suggested.) Also, please clarify what “maximum isometric force” (line 154) is, as opposed to “maximum isometric muscle forces” on line 153.
Experimental design: I agree with the authors’ statement, “the resent model is on the upper end of the complexity spectrum (line 362).”  However, one concern may be the sensitivity of the muscle fiber-type composition to the metabolic cost. The metabolic-cost computation in this study is based on a mathematical model developed by Umberger et al (2003) (modified by Koelewijn et al (2018)), which includes percent fast-twitch fiber type in the computation. Therefore, different fiber compositions in the model muscles would influence the computed metabolic cost. In fact, Umberger et al. (2006) (*1) who used the same method showed different metabolic cost with different fiber types in the simulation study of cycling. Some experimental studies have shown variability of fiber types in muscles in different individuals (e.g., Staron et al., (2000) (*2)). I suggest that the authors discuss the extent of influence of the fiber type variability on the study results.
*(1) Umberger et al. (2006) Journal of Biomechanics, 39: 1472-79
*(2) Staron et al. (2000) Journal of Histochemistry and Cytochemistry 48(5): 623-39
Validity of the findings: No comment.
Additional comments: The primary objective of the study is to compare the metabolic cost of walking between pre- and post transtibial amputation, using a 3D dynamic simulation approach. The results further confirmed the author’s previous work using 2D models, concluding that the transtibial amputation by itself would not cause an increase in metabolic cost. The strength of the study is demonstrated in the extensive work to generate simulations and verify the models and simulation results. (Testing with different fiber type compositions is recommended as stated in Experimental Design.) I think the study was conducted well to reach the conclusion, but I personally would like to know why or how some subjects increased post-amputation metabolic cost and others decreased as shown in Figure 9. If the authors can discuss what muscle group(s) contributed to the metabolic cost increase or decrease even in a few specific subjects, I think it would further enhance the value and quality of the study. If not in this paper, I certainly look forward to it in the future.


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: TRANSTIBIAL LIMB LOSS DOES NOT INCREASE METABOLIC COST IN THREE-DIMENSIONAL COMPUTER SIMULATIONS OF HUMAN WALKING
Review round: 1
Reviewer: 2

Output:
Basic reporting: Manuscript is well structured and generally clear. Some content might be transferred to supplementary material
Experimental design: Research question is well defined and methods are generally clear. Some minor issues regarding methods could be clarified and
Validity of the findings: The conclusions are sound and valid. Some additional analysis and conclusion could be added on the relation between the few observed kinetic and kinematics differences between pre-post conditions, how compensation for these differences occur and how they affect metabolic cost of walking
Additional comments: This simulation study investigates the metabolic energy cost of walking with a transtibial prosthesis and questions whether a post-limb loss gait would result in a different metabolic cost compared to pre-limb loss gait when initial musculoskeletal properties and body composition were maintained, besides from replacing the amputated limb by a passive transtibial prosthesis. This simulation study allows to further investigate the experimental observation that high functioning amputees do not seem to have greater energy cost than able-bodied controls. It seeks whether musculoskeletal mechanics allow a near normal gait pattern and non-significant differences in energy cost when the biological lower limb is replaced by a passive prosthesis (in the absence of other constraints such as balance control or stump-socket comfort).  It is a very interesting and relevant outcome that this hypothesis could be confirmed.
This is a very relevant study, using a state of the art modelling approach. The procedure is generally well described and the conclusion is generally sound. It is of interest to the scientific and clinical community.  Nevertheless, there are some issues that would deserve more attention to further enhance clarity and of the methods, results and interpretation.

Major comments:

-	The results demonstrate that metabolic cost does not differ significantly between simulations per and post limb loss and that deviations in the gait pattern are limited. Nevertheless, they indicate some deviations that are similar to experimentally obtianed data from amputee gait. Specifically, they observe reduced peak anterior GRF and reduced second vertical GRF peak on the prosthetic side. Next to some other small kinematic changes, of course plantar flexion of the prosthetic ankle was absent. It is of interest to further analyse to what extend these changes in gait kinetics and kinematics affect energy requirement and how this is compensated in the post-limb loss gait pattern.  For instance, it would be of interest to know how much mechanical energy was stored and released in the prosthesis, as this energy does not need to be generated by muscles at the expense of metabolic energy (hence this might reduce energy cost post-limb loss). Similarly, it is of interest how the reduced propulsive GRF (reduced anterior and vertical peak) affect mechanical and metabolic work? From a mechanical perspective the increased energy cost of prosthetic walking has been attributed to reduced push of which increased the energy required for step-to-step transitions (e.g. Kuo and Donelan Physical Ther 2010, Houdijk et all G&P2009). Did external mechanical work differ between pre- and post-limb loss simulations? or did joint (or muscle) work distribution differ between pre-post?  As the authors have the data to answer these questions, it would be interesting to expand the results section and discussion a bit in this direction.

-	The normalization of metabolic energy cost to body mass is an issue that deserves much attention in the results and discussion of the manuscript. While it is indeed not trivial which mass should be used for normalization and this is indeed often not well reported in literature, in this case I wonder whether any mass normalization would be required. The authors stress that they investigate the effects of limb-loss on metabolic cost with emphasis on only altering the mechanical properties of the lower limb, keeping all other musculoskeletal properties and body composition equal (and without taking into account other constraints such as balance and comfort). Hence, it is the same person with similar cardiorespiratory (aerobic) capacity that  walks with and without prosthesis. The true absolute energy consumption will be most relevant to him/her, regardless of which mass you divide it by. As you compare pre-post limb loss energy consumption within a participant, there is no need to normalize to body mass and absolute aerobic strain seems the best comparison.  (see also Wezenberg et al APMR 2012;94;1714-20.  Who use energy consumption normalized to aerobic capacity, i.e. relative aerobic load, as important factor in amputee gait)

-	In the cost function for optimization the authors include two terms; one related to deviations in the gait pattern and one related to energy cost. There is no explicit weighing between both I believe. Is this correct and would the results chance when the weighing between these two terms would be different?
It might be that the variable w is regarded as a weight factor. Although, it seems that that factor is used more to obtain realistic metabolic values from this approximated term for energy cost in the cost function, than that it is meant as a weight factor between the two terms in the cost function.

-	There are two tests for the validity of the model in the methods section (comparing the model to experiments with different masses to the leg and using an ankle brace). These validity tests are very relevant and adequate, but they are described very brief and the accompanying figures are rather prominent in the manuscript next to the main results of the study. Could these validations be transferred to supplementary material, where they can be described in little more detail and presented as addition experiment?

-	Figure 8-10 contain some overlapping information, one or two could be removed or merged. In my opinion

Minor comments:
-	Line 83-91:   several changes and additions were made to the original model of Dembia. Could you motivate why item 3-5 were modified/added? Was this crucial for this studies aim? Were these characteristic in the original model not valid?
-	Figure 2 is bit redundant as it only concerns one of many muscle groups in the model and it does not concern a main result of the study. Could be just in text references.
-	Line 137: does y in this equation only apply to the longitudinal displacement of stump relative to socket?
-	Line 143:  The methods and results of this analysis could be presented in supplementary material
-	Line 154:  maximum isometric force should be deleted from this sentence I guess, as it is already mentioned in the sentence before.
-	Line 160: I believe that a clinical relevant difference is more important than a minimal detectable difference. But no real consensus is available on clinical relevance difference on energy cost. Often 5-10% is used for walking in patients (no clear reference available on this)
-	Line 191: explain here that the output of the energy cost model used (Koelewijn 2018) cannot be used directly in the Moco software, and therefore you use this proxy equation. (now this is only clear at the end of the discussion line 372)
-	Line 192: can the excitation parameter t range between 0-1?
-	Line 194: explain the meaning of parameter w
-	Line 238: explain which initial guess is meant here (on which variables), how can these result from different values of w in the cost function? And why use the best out of three?
-	Line 262: when you talk about biological mass post-limb loss, do you mean the mass of the body with the mass of the amputated part of the leg, or the original biological mass before amputation?
-	Line 268: why use two one-sided tests and not one two-sided test?
-	Line 284: would AP impulse not be more relevant and reliable than peal anterior force, It is the impulse that relates velocity.
-	Line 291:  “few larger changes”  : which do you mean
-	Line 291: did you test the observed kinematic and kinetic differences statistically?
-	Line 355: IJmker et al did not conclude that energy cost of frontal plane balance is similar with and without transtibial limb loss, but suggested that constraints of the external stabilization system might conceal potential effects.  In both your 2D and 3D model you remove the need for ML balance control. Also the 3D model is ML stable because of tracking a cyclic motion including similar step width between models. As such you cannot investigate the effect of balance control in the 3D model and in that regards 2D and 3D model are equivalent I believe.
-	Line 422: I do miss some extra discussion on the few differences that exist in kinematics and kinetics between pre and post limb loss and how energy cost can be similar between both simulations despite these difference in gait pattern. Fig 4-6 present these kinematics and kinetics in detail but deserve limited attention in the text.
-	Fig 4: explain non-zero GRF in swing phase (most prominent in AP GRF)
-	Fig 5 knee extension differences in stance are remarkable and opposite than I would expect (I would expect moor knee extension in stance after limb loss, conform literature)


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: TRANSTIBIAL LIMB LOSS DOES NOT INCREASE METABOLIC COST IN THREE-DIMENSIONAL COMPUTER SIMULATIONS OF HUMAN WALKING
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall, this was good. I have one comment regarding figure 4, and a couple of possible typos:
1.	Figure 4: In both the pre- and post- simulation, there is ground contact after about 80% of the gait cycle, while the simulation should still be in the swing phase. Could you elaborate on what is happening there?
2.	Line 165 and 167: First you mention 26 subjects are required, and then 36 are created. Why the difference?
3.	Line 163: shouldn’t the third word be tests instead of test?
4.	Line 416-417: this sentence has “only” twice.
Experimental design: The experimental design is appropriate for the presented research question. I do have a couple of questions regarding the methods:
1.	Line 112-118: Why did you use the metabolic model developed by Koelewijn et al. (2018). I would not recommend to use this metabolic model for any post-simulation analysis. It was developed purely to be able to solve predictive simulations, and the outcome is different from the outcome of the original Umberger model, which is validated against data.
2.	Line 237-239: I would like to know a little bit more about the initial guesses that were used for the pre-op simulations, because the description was a bit vague here. It says that the initial guesses were from more preliminary simulations, but what preliminary simulations? And what were the initial guesses for these preliminary simulations?
3.	Line 148-159. What were these different ratios based on? Do I understand correctly that the total mass of the person was not altered? Why was that decision made? Why were the segment lengths not altered? I recently found that the effect on the final result is larger when changing the full body mass and length than the muscle parameters, so I was a bit surprised. Also, maximum isometric force and maximum isometric muscle force is mentioned twice. What is the difference?
4.	Line 187: why were the joint angles for which no (reliable) data was available not removed from the optimization instead of tracking 0 degrees?
5.	Line 130-144: Why is a double exponential function appropriate to model the socket motion? If this is presented, please add a justification. On the other hand, I am not sure if it really makes sense to mention this aspect so explicitly, when it is not actually used in the results.
6.	Line 191-198: I am a bit unsure about how you describe the choice of tracking weight here. If metabolic cost is the outcome parameter, it does not make sense to me to also only use metabolic cost to set the weight of the objective function. Then, you don’t know if maybe somehow two errors inside the simulations/analysis pipeline are cancelling each other. Since in my opinion an absolute comparison of the outcome of a metabolic energy model (only the modelled muscles + basal rate) to direct calorimetry measurements (which includes all muscles and other metabolic processes) does not really mean much anyways, this process is not convincing to me. However, the validation simulations are a good support for the choice of weights. Also, the kinematics do look realistic.
Validity of the findings: Overall, the presentation of the findings was good and meets the criteria.
1.	Section 3.3: I am not surprised that the damping values are so small, it is in my opinion surprising that they did not decrease to 0 exactly, because the higher the damping, the more energy is removed from the system, thus the higher the objective. However, this might actually be related to the tracking that is also included in the objective. Is there a way to actually compare the numbers to what is common in a current prosthesis? Numbers are presented for the stiffness in the discussion (line 376-391), but not for damping. I wouldn’t know where to find those myself.

I also have two comments regarding the supplementary data/code:
•	I changed line 267 to: path = [pwd filesep];% '/Users/rosshm/Documents/Matlab/TTA3D_solns/';
Then, the code should work when the current folder is TTA3D_solns for any computer.
You could probably even add a check to see if the user is in the right folder as follows:
curpath = pwd;
strcmp(curpath(end-10:end), ‘TTA3D_solns’);
•	If possible, it makes sense to me to also share to .osim files of the models. I did not find those in the data.
Additional comments: I enjoyed reading this paper and it is great that such a study can be performed with the MoCo software.


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: TRANSTIBIAL LIMB LOSS DOES NOT INCREASE METABOLIC COST IN THREE-DIMENSIONAL COMPUTER SIMULATIONS OF HUMAN WALKING
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is well written and clear
Experimental design: The research question is very relevant and well formulated. The design of the study is original and valid. Methods are clearly described and motivated in revised manuscript.

One small issue to address: While you basically test for equivalence in metabolic cost (based on research aim in introduction and statistical test used), you open the second paragraph of the discussion with  "This study was hypothesis-driven and addressed a research question with a yes/no answer, “Does limb loss increase metabolic cost?”   "
this should be adjusted.
Validity of the findings: Results and conclusions are valid within the general limitation of the simulation approach used. Potential effects of assumptions and simplifications in the simulation model are adequately discussed
Additional comments: Interesting study that provides valuable information and provokes relevant questions for further research.  Pleasure to 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: TRANSTIBIAL LIMB LOSS DOES NOT INCREASE METABOLIC COST IN THREE-DIMENSIONAL COMPUTER SIMULATIONS OF HUMAN WALKING
Review round: 2
Reviewer: 2

Output:
Basic reporting: The paper is clear and well written. I just have two small comments:

Line 285: a word appears to be missing
Line 346: "GRF appear" is used. I would use "GRF appears". When looking up if this was a plural abbreviation, I noticed that the abbreviation GRF was not defined in the paper. Therefore, please make sure that all abbreviations are defined.
Experimental design: Concerning one comment of the first review, I would recommend to also add this information to the manuscript itself:
Line 187: why were the joint angles for which no (reliable) data was available not removed from the optimization instead of tracking 0 degrees?

If there is no tracking target then the model tends to move these joints in unrealistic ways to track the other tracking targets unnecessarily well. Zero was used to minimize the range of motion of these joints, which are typically small.
Validity of the findings: No further comments.

It makes most sense to me to report the metabolic cost the way that it is done, because scaled metabolic cost makes the results comparable to experimental studies. However, it is interesting that the unscaled results could actually lead to a different conclusion.
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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an interesting study, and much work have been done to elucidate this topic.
While there are still much major and minor concerns for this study and manuscript.
Minor concerns:
1. Line 86, "This finding further was confirmed in CUMS rat model" was repeatedly.

2. Line 136, the last word should be "Guizhou" rather than "Guizbou".
Experimental design: 1. Why in the preclinical study, the female rats were selected as the animal model? how can author exclude the bias because of the menstrual cycle, especially in the female adults rats?

2. And totally 7 proteins were found differently between patients with MDD and healthy control, why author selected only two of them for the further study?

3. Why author selected the brain hippocampal tissues of SD rats to perform the Western blotting? How about the mPFC or other brain regions?
Validity of the findings: Author should add the examination of the other 5 proteins in the animal model, MYSM1, ITGB2, SMARCA2, RAB5C and PPP3CB. And Provide more evidences to support their discussion, for example, the 3 and 4 paragraph in the discussion.
Additional comments: 1. Line 86, "This finding further was confirmed in CUMS rat model" was repeatedly.

2. Line 136, the last word should be "Guizhou" rather than "Guizbou".

3. Why in the preclinical study, the female rats were selected as the animal model? how can author exclude the bias because of the menstrual cycle, especially in the female adults rats?

4. And totally 7 proteins were found differently between patients with MDD and healthy control, why author selected only two of them for the further study?

5. Why author selected the brain hippocampal tissues of SD rats to perform the Western blotting? How about the mPFC or other brain regions?


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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper was not well written with some details missing.
The overall results were weak.
The raw data were shared in public datasets.
Experimental design: For the RNASeq, the sample size is too small. Further, the lack of basic sample evaluations made the further judgment of their work difficult.
Validity of the findings: Weak connections between the validation rats and their RNASeq results.
Additional comments: Review of  Manuscript 56643, " Identification of key genes and crucial pathways for major depressive disorder using peripheral blood samples and chronic unpredictable mild stress rat models"

This paper analyzed the RNAseq data of blood samples from a total of 8 unique individuals, including 4MDD and 4 controls. A total of 247 DEGs, including 180 upgraded and 67 downgraded, were identified. These DEGs were annotated using GO and KEGG genesets. In addition, the DEGs were also put to String-db for network analysis. Using cytoscape cytoHubba plugin, 10 hub genes were identified and further annotated using David online annotation tools. Expression of these 10 hub genes was verified using qRT-PCR of blood samples from 50 MDD and 50 controls. Among these 10 genes, SQSTM1 was further studied and the protein level of SQSTM1 was shown to be positively correlated with TNFa, an inflammation marker with a sample size of 44 MDD and 44 controls. Finally, in the brain, SQSTM1 were also found to be overexpressed in CUMS rats.
Before publications, there are still a lot of questions that need to be addressed. The most important results of this paper came from DEG analysis of the blood of human subjects, with only 4 samples per group. Given the high diversity of human subjects, these results were not reliable especially that the authors wanted to use the gene expressions in the blood to study a brain disorder.
Besides, there are some other technical details that need to be clarified.

Main comments:
1.	Please add a table describing the basic demographic information of the key 8 human subjects. This table should include the information of sex, age(or age range), medicines, etc. Since only 4 subjects per group, the review wanted to exclude the possibilities of the effect from other covariables.
2.	From Figure 1A, it seems that their results were driven from MDD3. A PCA plot of the 8 subjects may help to check whether MDD3 is an outlier or not.
3.	In Figure 1, the log2fold change of most DEGs were above 10. This means a 1000 fold increase. This is unlikely based on the heatmap in A. Please check. Also, in 1A, please add a legend of the values in the heatmap. If it was the gene expression, please add whether it is the raw counts or lcpm, or something else. In the related method part, please add details on how the sequencing data were processed.
4.	In Figure 2 and Figure 3C, Please add p-value information.
5.	Related to Figure 3B, the details of how the 10 hubs were selected should be described, including the rules of the thresholds, and the basic algorithms of the plugin.
6.	Colors in Figure 3A is not described. Also, in String-db, the thresholds of the chosen PPI were not described.
7.	Why only SQSTM1 were selected for further studies?


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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall, the paper is written in good format with minor editing in the languages. The reference are properly used.
Experimental design: The experimental design has sufficient amount of biological replicates and the data has been analyzed and validated using various techniques. There are some gaps that need to be filled. Please find the details in the attached documents.
Validity of the findings: The authors need to provide the raw files for the sequencing data. The fold change in the sequencing data needs to be shown and along with some background about the clinical data to appreciate the differences seen in figure 1.
Additional comments: Overall it is well thought out plan. The authors need to address some of the comments to strength the manuscript.

The authors have a good number of samples for their RNA sequencing data, validation, and rat model data. However, there are some blanks that need to be filled to strength the findings.
•	TNF-α needs to be questioned in the rat model (either using IF or western blot)
•	Need to validate the inflammatory response caused by TNF-α using the samples from rat model or patients.
•	Role of SQSTM1 in other mental health disorders has been established.
•	Some of the other genes such as AKT2, TSC2, RAB5C, ITGB2 are involved in autophagy and phagocytosis need to be validated in the rat model or in the human samples using rtPCR to strengthen the manuscript.
•	There are minor edits that needs to be addressed to 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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 2
Reviewer: 1

Output:
Basic reporting: In the revised manuscript and response letter, the authors addressed most of my questions. The remaining concern is about the data availability policy, which would make it less efficient for repeats and comparisons.
Something minor things still need to be addressed:
1.The original question 6: Still need to provide detailed descriptions about the cut-offs of your PPI. The score cut-off is to limit the number of interactions. Setting different cutoffs will affect your final results.
2." The algorithms applied to identify hub genes were MCC and Degree", what is "MCC" short for? Could the author briefly describe the algorithms in the revised version?
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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors written the manuscript well and included all the raw data
Experimental design: The experiment design is accurate to test and validate the hypothese
Validity of the findings: The findings have been validated by various techniques which has helped strengthen the 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: IDENTIFICATION OF KEY GENES AND CRUCIAL PATHWAYS FOR MAJOR DEPRESSIVE DISORDER USING PERIPHERAL BLOOD SAMPLES AND CHRONIC UNPREDICTABLE MILD STRESS RAT MODELS
Review round: 3
Reviewer: 1

Output:
Basic reporting: In the revised version and the response letter, the author addressed my questions.
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: CELLULOSE AEROGEL COMPOSITES AS OIL SORBENTS AND THEIR REGENERATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article introduces interesting findings about cellulose aerogel composites as oil sorbents. These are not very breakthough ones but meaningful for industry and can be good reference for other studies. Literature references and background are sufficiently provided.
Experimental design: The research question is well defined and the method described with sufficient information. But more details should be provied if possible (see the general comments)
Validity of the findings: Please review the Figures 6 to 17. Each figure only has part (a) (or part (b)). And the caption of some figures are the same, such as figure 16 and figure 17.
Additional comments: - Can the author explain the measurement of the water contact angle in more details. For example, which device was used to measured the angle? How did it calibrated?
- In the graphs (Fig.6 to Fig.17),the legend can be put into white backrgound text box to avoid being cut through by the grid lines
- Line 206, please check the range of contact angles 120-125 degree. In the next line, the authors mention that the highest is 137.8 degree?
-


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: CELLULOSE AEROGEL COMPOSITES AS OIL SORBENTS AND THEIR REGENERATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: English should be improved for the clearly understanding of the paper.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The manuscript described the study on preparation of cellulose aerogel composites for oil sorbents application. The data is interesting and is suitable to the journal. However, it need further experiments and data so that the content of the research is clarified and the explanation of obtained result is more proper.

1. Specific surface area measurement should be carried out. It helps to explain the adsorption of the materials more clearly than the porosity solely does.
2. Stability of aerogel when oil was absorbed in the sample should be investigated. For example: evaluate the mechanical properties of sample after each time of absorption.
3. SEM image should be provided to clearly observe the structure as well as the surface of sample. This may be helpful in the explanation of contact angle because contact angle was predominantly dependent on the surface structure.


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: CELLULOSE AEROGEL COMPOSITES AS OIL SORBENTS AND THEIR REGENERATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: the content of this article is clear and well-written.
Experimental design: method has been sufficiently updated
Validity of the findings: no comment
Additional comments: The authors provide acceptable answers for the questions and comments. The paper is recommended to be published on this journal.


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

Input:
Title: CELLULOSE AEROGEL COMPOSITES AS OIL SORBENTS AND THEIR REGENERATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors revised the manuscript according to the reviewer's comments. The manuscript is acceptable.


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: A COMPARATIVE STUDY ON KNOWLEDGE TOWARDS COVID-19 PREVENTION AMONG UNDERGRADUATE STUDENTS IN MACAO AND ZHUHAI, CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: This is a survey regarding to COVID-19 preventive knowledge, which aimed to compared COVID-19 preventive knowledge of undergraduates between Macao and Zhuhai. Though this is an interesting study, I have some comments for the authors for their consideration.
1. The language of this manuscript need to be improved.
2. Line 17-18. Please refine the conclusion. The initial conclusion is not accurate.
3. Line 38, 48. Please re-check the format of the citation.
4. For the COVID-19 preventive knowledge inventory, please add the information about psychometric properties.
5. Line 92, authors should calculate the response rate.
6. Line 86, please specifically describe the covariates.
7. Please use “<0.001”to replace “0.000”.
8. why did you use cut-off of 20 in the age? Please provide the reasons.
9. Table 4. For the footnote, please use “female” rather than “Feale”
10. Major status is a good index. Hence, authors should further study the difference between medical students and non-medical student.
11. Only three covariates were presented, do you have any more socio-demographics?
12. Authors did not compare the difference of the findings in between previous studies and this study in the discussion. Please add them.


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: A COMPARATIVE STUDY ON KNOWLEDGE TOWARDS COVID-19 PREVENTION AMONG UNDERGRADUATE STUDENTS IN MACAO AND ZHUHAI, CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. English language of the paper is not so good. Please have it polished by English-speaking specialists after the revisions, including punctuations. For example, line 5-6, “difference” and “different” seem to be repeated. Line 17 “the certain points of” is an unclear term.

2. Line 38 and line 48, please check the reference “MOE-China” and “DSEDJ-Macao” represent what?

3. Introduction. This part is written well. I think the authors may consider to emphasize the interplay of COVID-19 infection because the two cities are bordering and university students can visit each other very conveniently. I remember the two cities opened to each other since August 2020. In this context, investigating COVID-19 knowledge of the two cities has important public health significance.

4. Line 54, please add title “Methods”.

5. Line 65, please provide the full form of “MUST”.

6. The authors dichotomized age at 20 years, please specify why.

7. Are there any i8nternational students participated in the survey, in particular Macau, an international city. The authors may consider to describe the inclusion and exclusion criteria.

8. In discussion part, the authors mentioned peer health education for implications, but this would result in high risk of COVID-19 transmission.
Experimental design: 1. English language of the paper is not so good. Please have it polished by English-speaking specialists after the revisions, including punctuations. For example, line 5-6, “difference” and “different” seem to be repeated. Line 17 “the certain points of” is an unclear term.

2. Line 38 and line 48, please check the reference “MOE-China” and “DSEDJ-Macao” represent what?

3. Introduction. This part is written well. I think the authors may consider to emphasize the interplay of COVID-19 infection because the two cities are bordering and university students can visit each other very conveniently. I remember the two cities opened to each other since August 2020. In this context, investigating COVID-19 knowledge of the two cities has important public health significance.

4. Line 54, please add title “Methods”.

5. Line 65, please provide the full form of “MUST”.

6. The authors dichotomized age at 20 years, please specify why.

7. Are there any i8nternational students participated in the survey, in particular Macau, an international city. The authors may consider to describe the inclusion and exclusion criteria.

8. In discussion part, the authors mentioned peer health education for implications, but this would result in high risk of COVID-19 transmission.
Validity of the findings: 1. English language of the paper is not so good. Please have it polished by English-speaking specialists after the revisions, including punctuations. For example, line 5-6, “difference” and “different” seem to be repeated. Line 17 “the certain points of” is an unclear term.

2. Line 38 and line 48, please check the reference “MOE-China” and “DSEDJ-Macao” represent what?

3. Introduction. This part is written well. I think the authors may consider to emphasize the interplay of COVID-19 infection because the two cities are bordering and university students can visit each other very conveniently. I remember the two cities opened to each other since August 2020. In this context, investigating COVID-19 knowledge of the two cities has important public health significance.

4. Line 54, please add title “Methods”.

5. Line 65, please provide the full form of “MUST”.

6. The authors dichotomized age at 20 years, please specify why.

7. Are there any i8nternational students participated in the survey, in particular Macau, an international city. The authors may consider to describe the inclusion and exclusion criteria.

8. In discussion part, the authors mentioned peer health education for implications, but this would result in high risk of COVID-19 transmission.
Additional comments: 1. English language of the paper is not so good. Please have it polished by English-speaking specialists after the revisions, including punctuations. For example, line 5-6, “difference” and “different” seem to be repeated. Line 17 “the certain points of” is an unclear term.

2. Line 38 and line 48, please check the reference “MOE-China” and “DSEDJ-Macao” represent what?

3. Introduction. This part is written well. I think the authors may consider to emphasize the interplay of COVID-19 infection because the two cities are bordering and university students can visit each other very conveniently. I remember the two cities opened to each other since August 2020. In this context, investigating COVID-19 knowledge of the two cities has important public health significance.

4. Line 54, please add title “Methods”.

5. Line 65, please provide the full form of “MUST”.

6. The authors dichotomized age at 20 years, please specify why.

7. Are there any i8nternational students participated in the survey, in particular Macau, an international city. The authors may consider to describe the inclusion and exclusion criteria.

8. In discussion part, the authors mentioned peer health education for implications, but this would result in high risk of COVID-19 transmission.


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: A COMPARATIVE STUDY ON KNOWLEDGE TOWARDS COVID-19 PREVENTION AMONG UNDERGRADUATE STUDENTS IN MACAO AND ZHUHAI, CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: Thank you for your revision. Only one minor error need to be addressed.
1. "In order to strategy more suitable campus-based coronavirus disease 2019 (COVID-19) " . "Strategy" is a noun rather than verb.


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: A COMPARATIVE STUDY ON KNOWLEDGE TOWARDS COVID-19 PREVENTION AMONG UNDERGRADUATE STUDENTS IN MACAO AND ZHUHAI, CHINA
Review round: 2
Reviewer: 2

Output:
Basic reporting: See my comments as below.
Experimental design: See my comments as below.
Validity of the findings: See my comments as below.
Additional comments: The manuscript has been substantially improved. However, I still found several language problems in this version, and listed some as below. Please check and re-edit the paper all throughout
1. Title should be added "China".
2. Abstract. Line 12, "strategy" is not a verb which can not be used in this way. Line 19-20, the two sentences are problematic. Line 21, SARS-CoV-2 and COVID-19 virus are duplicated, which one the authors prefer?
3. Line 54, what is GBA?


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: DOG1 IS COMMONLY EXPRESSED IN PANCREATIC ADENOCARCINOMA BUT UNRELATED TO CANCER AGGRESSIVENESS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall, the manuscript writing is clear and easy to understand. Also, the results are concise. With regard to reporting, I listed a few comments to further improve manuscript readability as seen below:

Line 72-73: this part is trying to summarize the reported cases of DOG1 across multiple human organs and cancer types, yet it is a bit unclear what exactly are the organ-cancer pairs given the current wording.

Line 80: "From data derived from the TCGA database". Recommend a rewrite. Also in line 80, do "these authors" refer to Crottès et al?

Line 86-90: Recommend to split into 2-3 sentences for better readability.
Experimental design: To account for confounding variables in the analysis, it might be better to add other metadata such as sex, age, smoking status, and also include the correlation test p-value (with pancreatic cancer).
Validity of the findings: Overall a straightforward study. A suggestion regarding the evidence supporting key finding: more details are needed to support the key result that "DOG1 gene expression is unrelated to pancreatic cancer aggressiveness". E.g. What are the exact clinico-pathological parameters used for the correlation test? Is it catogorical variable or continuous variable? Is the Chi-squred test applicable to the data types?
Additional comments: Cancer genomics/transcriptomics, and biomarker identification for pancreatic cancer are important topics, and we welcome the work from the authors to contribute. There are some areas outlined above that could be improved for better manuscript quality.


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

Input:
Title: DOG1 IS COMMONLY EXPRESSED IN PANCREATIC ADENOCARCINOMA BUT UNRELATED TO CANCER AGGRESSIVENESS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this manuscript, the authors highlight the association of DOG-1 with pancreatic adenocarcinoma but not it's aggressiveness. The primary method of analyzing DOG-1 is immunohistochemistry of microarray samples followed by statistical analysis. The article is written in clear English. Tables and one figure is clear. Literature references seem to be in order. I have few general comments/questions-
1. Why did the authors not skip the non-informative samples completely? If only 501 out of 599 were analyzable then why not skip them? If the authors wish to include this anyhow, it is not a good idea to include this information as an independent result, especially not the first one.
2. Proof-reading for typos will help 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: DOG1 IS COMMONLY EXPRESSED IN PANCREATIC ADENOCARCINOMA BUT UNRELATED TO CANCER AGGRESSIVENESS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have responded satisfactorily to my comment. Minor typos have also been addressed. I congratulate the authors on their work.


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: IMPACT OF THE THE COVID-19 PANDEMIC ON A QUEEN CONCH (ALIGER GIGAS) FISHERY IN THE BAHAMAS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article seeks to evaluate how an increase in fishing pressure, on a population of queen conch, during the introduction of COVID-19 control measures effects a rural small-scale fishery in The Bahamas.
The article is well written and suitable references are used and good background information is given.
Figures and tables are sufficient and raw data is available.
The results may be reasonably predictable, given the circumstances, but demonstrate how just moderate increases in fishing pressure on a small population of easily fished conch can have a serious impact in a short period of time.
Experimental design: The work seeks to use the situation brought about by the global pandemic and subsequent restrictions imposed in many communities to be able to evaluate the impact upon the conch fishery when subsistence fishing is one of a few activities allowed.
Although this does create a unique opportunity the results could most likely be predicted, although the question remains well define and is potentially meaningful.
Methods are described well and could be repeated.
Validity of the findings: Data is robust and the conclusions are well stated and acceptable
Additional comments: I think it would be helpful are the start to just define what subsistence fishing is, say with respect to artisanal fishing. You touch on this later but some guidance at the start would help in my opinion.
Also consider some wider referencing outside of The Bahamas, for example Tiley et al 2018 suggest that lip thickness should also be used for juvenile conch
J Invertebr Pathol 2018 Jun;155:32-37.
 doi: 10.1016/j.jip.2018.04.007. Epub 2018 Apr 24.
Pathology and reproductive health of queen conch (Lobatus gigas) in St. Kitts


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: IMPACT OF THE THE COVID-19 PANDEMIC ON A QUEEN CONCH (ALIGER GIGAS) FISHERY IN THE BAHAMAS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1-Basic reporting

-Clear and unambiguous, professional English used throughout. Literature references, sufficient field background/context provided.

	Professional English used through, let references and background could be improved.

-Professional article structure, figures, tables. Raw data shared.
Self-contained with relevant results to hypotheses.

Reporting is clear and well written and supported to the extent possible in the underreported topic of subsistence fishing.  The structure of the article is professional and raw data is shared.
Experimental design: 2-Experimental Design

-Original primary research within Aims and Scope of the journal.
Subsistence fishing data is rare in the Bahamas so this is unique snapshot of such data collected opportunistically after a national disaster type event.  It could be considered to be original research but there is no hypothesis as this is more of a descriptive study.

-Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.

L 66– The lead sentence of the paragraph and the concluding one give the general purpose, but the paragraph should be a bit more succinct.

-Rigorous investigation performed to a high technical & ethical standard.
	This descriptive study collected that best data available at the time.

-Methods described with sufficient detail & information to replicate.

-A figure with a map of the study area location including its extent and the location of the middens by number would be useful.

-How they measured conch should be explained more detailed than just "standard morphometric measurements". One of the papers by Appeldoorn or other source should be referenced.  In particular, the method of measuring LT should be defined as method can easily impact measurements.

-The reader needs a better understanding of the rationale for the conch size classifications other than just matching the earlier study.  For clarity, add size category initials (from Clark et al. and raw data file) to the legend on Fig. 4.

-The topic of legal harvest vs. sexual maturity should be brought up in the Methods section.  Otherwise, it's not really clear why the SL and LT groups are presented as they are in Fig. 4.

-Other literature to be consulted include:  Stoner, AW, Mueller, KW et al.  Maturation and Age of Queen Conch:  Urgent need for changes in harvest criteria. Fish. Res., 131: 76-84 (2012).  Also a technical paper for the Bahamas Ministry of Ag and Marine Resources entitled, “Queen Conch Harvest in The Bahamas during the Last Decade: Shell Middens Provide More Evidence of Overfishing.”

-L 80-81 “Discarded shell middens are assumed to be an accurate representation of fisheries landings.” My interpretation of the referenced paper does not give support for that statement and based on personal observations, there are numerous shells left under the water.
Validity of the findings: 2-Experimental Design

-Original primary research within Aims and Scope of the journal.
Subsistence fishing data is rare in the Bahamas so this is unique snapshot of such data collected opportunistically after a national disaster type event.  It could be considered to be original research but there is no hypothesis as this is more of a descriptive study.

-Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.

L 66– The lead sentence of the paragraph and the concluding one give the general purpose, but the paragraph should be a bit more succinct.

-Rigorous investigation performed to a high technical & ethical standard.
	This descriptive study collected that best data available at the time.

-Methods described with sufficient detail & information to replicate.

-A figure with a map of the study area location including its extent and the location of the middens by number would be useful.

-How they measured conch should be explained more detailed than just "standard morphometric measurements". One of the papers by Appeldoorn or other source should be referenced.  In particular, the method of measuring LT should be defined as method can easily impact measurements.

-The reader needs a better understanding of the rationale for the conch size classifications other than just matching the earlier study.  For clarity, add size category initials (from Clark et al. and raw data file) to the legend on Fig. 4.

-The topic of legal harvest vs. sexual maturity should be brought up in the Methods section.  Otherwise, it's not really clear why the SL and LT groups are presented as they are in Fig. 4.

-Other literature to be consulted include:  Stoner, AW, Mueller, KW et al.  Maturation and Age of Queen Conch:  Urgent need for changes in harvest criteria. Fish. Res., 131: 76-84 (2012).  Also a technical paper for the Bahamas Ministry of Ag and Marine Resources entitled, “Queen Conch Harvest in The Bahamas during the Last Decade: Shell Middens Provide More Evidence of Overfishing.”

-L 80-81 “Discarded shell middens are assumed to be an accurate representation of fisheries landings.” My interpretation of the referenced paper does not give support for that statement and based on personal observations, there are numerous shells left under the water.
Additional comments: This study was undertaken as a rapid review of subsistence fishing of queen conch in The Bahamas in response to COVID-19 restrictions and as such, is unique.  It is generally well researched and developed into the context of the existing literature (but see note below on literature that needs to be consulted).
The manuscript borders between being a research paper and a case study and as such could be reduced to a shorter note.


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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is trying to elucidate the role of DUSP12 gene and hepatocellular carcinoma.
Experimental design: The overall design of this study is comprehensive. The report of the methods is robust.
Validity of the findings: The validity of the results is reliable. The findings and conclusions were well supported.
Additional comments: This article is trying to elucidate the role of DUSP12 gene and hepatocellular carcinoma. The overall design of this study is comprehensive. The report of the methods is robust. The validity of the results is reliable. The findings and conclusions were well supported. Thus, I suggest accepting this paper.
Figure 4, for survival analysis, please specify how do you separate high and low expression? Please also add a sample number for each group in figure 4C-F.
1.	Please increase the font size in figure 1, 5, 6, and 8.
2.	Figure 8a and 8b, please set a threshold for correlation analysis, combine correlation coefficient and P values.
3.	Please transfer figure S3A, S3B, and S3c to figure 9 or 10.


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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Ju et al, propose the role of DUSP12 in the pathogenesis of HCC. The authors use an extensive and data-rich approach to support their findings. I find the data and the model of high clarity. I would support the publication of this interesting concept if the authors could address some important comments.

First, the language used in clear, a bit harsh in some sections. The authors discuss the existing literature satisfactory.
Experimental design: Although the link between DUSP12 and HCC is biased (not supported by an unbiased retrospective approach), analyses used are well controlled, explained and extensively described. I would like the authors to clarify if the p values in figure 8 are adjusted or not.

I have a couple of additional suggestions experimental-wise that will be explained in the following section.
Validity of the findings: Although the model proposed from the authors is solid and thorough, I would like to propose some additional analyses.

1. The authors could analyze the cell line dependency to DUSP12, using the Achilles dependency project of the Broad institute https://depmap.org/portal/
if this model shows dependency of HCC cell lines to DUSP12 in an unbiased way, it will go a long way on supporting the proposed model

2. The authors could also use SNE plots to show the cluster that express DUSP12 in their scRNA seq data.

3. It would be extremely interesting to include a table with the top positively and negatively correlated genes in TCGA dataset for DUSP12.
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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 1
Reviewer: 3

Output:
Basic reporting: Reporting looks to be in good form.
Experimental design: The connection between DUSP12 and HCC is carefully explored in this paper and represents original primary research suitable for this journal.
Validity of the findings: Analyses were conducted with multiple modalities -- multiple online databases and loss of function assays.

Conclusions are well stated -- not overly stated -- and limited to supporting results.
Additional comments: A comprehensive analysis was conducted investigating the basic science and translational properties of DUSP12 as it relates to HCC. This seems to be a meticulously prepared research report and I do not have any criticisms.


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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 1
Reviewer: 4

Output:
Basic reporting: In the manuscript (#61090) entitled, “DUSP12 regulates the tumorigenesis and prognosis of hepatocellular carcinoma”, the authors have have aimed to evaluate the role of DUSP12 in tumorigenesis, infiltration of immune cells, and prognosis of hepatocellular carcinoma (HCC). Ju et al have utilized a variety of online tools, multiple online databases, and loss of function assay to evaluate the role of DUSP12 in HCC. The authors however have failed to use a well-planned and comprehensive approach to come up with a logical conclusion and the manuscript is not well written.
The data presented/experiments performed are not rigorous enough and does not justify the conclusions.
The major drawback of the manuscript is that it is written very vaguely, making it difficult to comprehend. Authors in many places through the manuscript have just written down speculations without any concrete evidence and have failed to comprehensively explain their own findings.
Figure legends merely indicate the technique/graph and are not informative or comprehensive at all.
A very important issue about this manuscript is the language. There are numerous grammatical errors, typographical errors, punctuation mistakes, confusing sentences and a peculiar choice of words in many instances making the comprehension of data difficult. Hence the authors should revisit the manuscript and edit it thoroughly preferably with the help of someone with command in English language.
Experimental design: Results are just an extension of Materials and Methods and do not explain the findings.
Discussion is merely a summarization of results and hardly manages to explain the findings making it extremely difficult to the reader to comprehend the relevance/rationale of the findings and techniques used.
The name of the cell line Huh7 is represented in the manuscript differently in different places (Huh-7), needs homogenization. Also, can the authors comment on using Huh7 cell line for their experiments, although as per their data (Figure 3C) it is on the lower end of spectrum for DUSP12 expression.
There are a lot of question marks regarding the data throughout the manuscript which I have tried to detail, hence, making it impossible to the reviewer to be confident about the findings put forward by the authors.
Validity of the findings: Figure 1C,D,E: The number of samples in stage 4/grade 4/N1 are significantly less as compared to others and hence there is no significant change in those, authors fail to acknowledge/explain this.
Figure 3A,B: No mention of the technique used to get these images/results anywhere in the manuscript.
Figure 6B: No mention of number of samples/replicates.
Figure 7D: MCODE analyses, the reviewer is unclear about the relevance and significance of this and the authors have made no attempt in the manuscript to comprehensively explain/justify it.
The authors need to revisit their data and try to make logical conclusions in a more comprehensive manner based on their original findings rather than just putting down speculations.
Additional comments: 1.	Formatting errors throughout the manuscript (even in sub-headings)- Line 48 (function not functional), Line 100 (TNF-alpha?), Line 223 (protein), Line 264 (TGF-beta?), Line 268 (different), Line 277 (motility).
2.	Spacing errors Line 86, 163, 239.
3.	Grammatical errors Line 89 (and is?), Line 103-104, Line 132 (was downloaded), Line 161 (provides), Line 164 (were utilized), Line 173 (Huh7 was), Line 205 (After removed the cells?).
4.	Line 90-91: “The median duration of survival of patients with advanced HCC is 1 year, do the authors mean Chinese population? as the reference used is based on data from a Chinese medical center (Zhu et al 2015).
Line 95-96: Order in which the references are cited is incorrect.
Line 153: A total of 369 tumor samples extracted from the GDC, Line 155 A total of 130 of 261 tumor samples, unclear?
Line 247: 392 DEGs mentioned, where as Line 141 mentions only 369.
Line 269-272: The sentence is unclear and does not make sense in its current form, needs formatting.
Line 282: The authors propose this not infer as the manuscript does not have confirming data, also the role of DUSP12 has been previously mentioned in the manuscript and hence redundant here.
Line 298: Again the statement is more of a speculation than conclusion of results.
Line 313: MCODE analyses, authors justb explain 1 and 2, with no justification/mention of others.
Line 325: We showed that Uusp12 could recruit Tregs to infiltrate liver-cancer tissues, how, where is the data/explanation to this?


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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: No new comments.
Experimental design: No new comments.
Validity of the findings: No new comments.
Additional comments: No new 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: DUSP12 REGULATES THE TUMORIGENESIS AND PROGNOSIS OF HEPATOCELLULAR CARCINOMA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The changes made by the authors in regard to the comments by reviewers has made the manuscript more comprehensible than before and has improved the general writing and language. Results are clearer now.
Experimental design: New changes to the methods/results section makes the experimental approach clearer and understandable to the reader.
Validity of the findings: The results of the study are beneficial for the field and concluded correctly.
Additional comments: The editing has made the manuscript much better and more comprehensible. I am happy with the revision.


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: ASPIRIN INHIBITS TUMOR PROGRESSION AND ENHANCES CISPLATIN SENSITIVITY IN EPITHELIAL OVARIAN CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: Throughout the text, there are grammatical errors.  Please make sure that the sentences are scientifically sound and grammatically correct.
More updated literature can be cited.
Experimental design: The experimental design is appropriate. Methods described are sufficient.  Rigorous investigations performed.
Validity of the findings: The results presented are interesting and conclusions well written.
Additional comments: It is reported that ovarian cancer is a common gynecological malignancy, and it is difficult to treat it because these cancers often exhibit chemoresistance.  The authors in this paper studied the effect of aspirin alone or in combination with cisplatin (CDDP) on epithelial ovarian cancer cells (EOC) cell proliferation, cell migration, colony formation, apoptosis etc. The investigated cell lines include: A2870, Caov-3 and SK-OV3.  The authors demonstrated that aspirin treatment inhibited the migration, proliferation in these cells and also induced apoptosis.  Interestingly, aspirin potentiated the effect of CDDP.   The authors demonstrated that aspirin induces acetylation of p53; however, does not affect the expression levels of p53.  This was associated with increased expression of p21, Bax, Foxf1, PUMA and Rad proteins in these cell lines.  They conclude that aspirin’s action on tumor regression and enhancement of CDDP may occur through acetylation of p53 and induction of p21, Bax, etc.

Comments:
1.	The authors state that the A2780 and SK-OV-3 cells contain wild type p53 and Caov-3 cells contains mutant p53.  It appears that from the results presented, aspirin acetylates both wild type and mutant p53.  What is the p53 mutation (amino acid replacement) reported in Caov-3 cells?
2.	P53 contains several lysine residues that are potentially acetylated by aspirin.  The antibodies used in the study detected K382.  It will be useful to discuss the significance of K382 acetylation for gene expression if that information is available in literature.
3.	 The authors emphasized on the potential role of p53 acetylation in aspirin’s anti-cancer effects.  They should open to the possibility that other pathways may also contribute to aspirin’s anticancer effects.  In this regard, citing a recent review article published by Sankaranarayanan et al (International Journal of Molecular Sciences, Int. J. Mol. Sci. 2020, 21(23), 9018; https://doi.org/10.3390/ijms21239018 is appropriate.
4.	 Throughout the text, there are grammatical errors.  Please make sure that the sentences are scientifically sound and grammatically correct.


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: ASPIRIN INHIBITS TUMOR PROGRESSION AND ENHANCES CISPLATIN SENSITIVITY IN EPITHELIAL OVARIAN CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: The manuscript entitled “Aspirin inhibits tumor progression and enhances cisplatin sensitivity in epithelial ovarian cancer” investigated the effects of aspirin on several ovarian cell lines and cisplatin-treated cells. Besides, its tumor effect on tumor-bearing rodent was evaluated. Finally, the acetylation status of p53 was examined. In conclusion, aspirin can inhibit tumor progression and enhance the CDDP sensitivity of EOC cells. These antitumor effects of aspirin might be mediated by p53 acetylation and subsequently activation of p53 target genes.

Besides its effect on COXs, antitumor effect of aspirin is well reported in many malignancies, including ovarian cancer. Not only aspirin but also aspirin derivatives are demonstrated to have antitumor effects and sensitize tumor cells to chemotherapeutic treatments including cisplatin. Thus, current study focuses and findings are not new to the readers.

1.	There are studies demonstrating the effects of NSAIDs, aspirin, and aspirin derivatives on malignancies, including ovarian cancer. Unfortunately, those relevant studies and findings were not described and mentioned in this manuscript.
2.	Figure legends should be checked for the clear description of experimental protocols and conditions.
3.	Figure 2 and figure 3, a group of aspirin alone should be included for comparison.
4.	Current study only measured the change of p53 acetylation after treatment in the presence of aspirin, well-known phenomena. The specific involvement and action of p53 should be determined.
5.	Overall, current findings were not enough to provide additional insights into the study of aspirin and ovarian cancer. Advanced studies are needed.


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: EMBRYO AND LARVAL BIOLOGY OF THE DEEP-SEA OCTOCORAL DENTOMURICEA AFF. METEOR UNDER DIFFERENT TEMPERATURE REGIMES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript fulfils the criteria for basic reporting, with only minor adjustments needed.
Experimental design: The experiment is well defined and performed. I have only one concern, and that is that only one male was used for the reproduction producing the larvae for the experiment. It worked out fine despite this, but it is a bit risky since not all couples produce viable offspring. This is just a note to the authors that they might want to consider using several individuals from each sex for future experiments.
Validity of the findings: No comment, all good.
Additional comments: Congratulations to a successful rearing of deep-sea coral larvae and subsequent experiments. The results are much needed!


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: EMBRYO AND LARVAL BIOLOGY OF THE DEEP-SEA OCTOCORAL DENTOMURICEA AFF. METEOR UNDER DIFFERENT TEMPERATURE REGIMES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Please, see the attached document.
Experimental design: Please, see the attached document.
Validity of the findings: Please, see the attached document.
Additional comments: Please, see the attached document.