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: HMGB1-ACTIVATIED NLRP3 INFLAMMASOME INDUCES THROMBOCYTOPENIA IN HEATSTROKE RAT
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript conforms to the guidelines of the journal. Figure legends contain special characters (perhaps asterisks?) that have not come through in the PDF for review and are shown as boxes. This should be corrected. Additionally, figure legends should clearly state the sample size for each experiment, including both independent experiments and technical replicates in order to conform with usual standards and make the figures interpretable.
Experimental design: Methods and experimental design are relevant and well described.
Validity of the findings: The authors submit and interesting set of studies regarding the role of inflammasome activation in the development of thrombocytopenia in a rat model of heat stroke. This is a relevant problem which merits study. The authors show that induction of heat stroke (HS) in rats results in thrombocytopenia and platelet activation (CD62P expression). They demonstrate that in HS, platelets up regulate NLRP3 and Asc, and that cleaved caspase-1 levels are increased as well. Blockade of NLRP3 with MCC950, as expected, decreases cleaved caspase-1 levels, but also down regulates IL-1B expression. HMGB1 levels were elevated in HS, and blockade of HMGB1 with ethyl pyruvate (non-specific) or monoclonal antibody reduced NLRP3 expression and cleaved caspase-1 in platelets. This effect was found to be both via TLR4 and RAGE, and led to increased ROS production in platelets. While these findings are of interest, I have several reservations related to both mechanistic concerns and synthesizing these data into an overall model.

1. HS induces a decrease in platelet count, as shown. However, the authors discuss in discussion that HS can induce platelet clustering. This is not unexpected in a condition of significant inflammation and platelet activation. Mechanistically and clinically, an absolute loss of platelet number is not the same as platelet clumping. Did significant platelet aggregation occur here? This could be assessed by examining a slide from the peripheral blood, or even by simple spectrophotometry of the platelet-rich plasma to differentiate between cell loss and cell clumping.

2. The electronic microscopy images are very nice and illustrate interesting ultrastructural changes in thee platelets in HS. However, these data are really not integrated into the overall manuscript. The images are described, but the significance of these findings in the context of inflammasome activation or HS are not mentioned at all. If they cannot be linked to the system being studied, I would suggest they be removed.

3. My next concern is in regard to much of the inflammasome activation data and interpretation. In general, inflammasome activation requires 2 steps (see many reviews on the topic). Step 1, priming, is generally mediated by TLR4 and results in the upregulation of inflammasome machinery (NLRP3, Asc, pro-IL-1B and pro-caspase-1). Signal 2, which can take on many forms (membrane damage, potassium depletion, ionophores, and others) then triggers the inflammasome, resulting in Asc clustering, pro-caspase-1 cleavage, and subsequent cleavage of pro-IL-1B and gasdermin D both to active forms. These two events can be separated temporally. Further, in some conditions, the resulting trigger will result in cell lysis via pyroptosis. The data shown in this manuscript seem to relate mostly to priming. Upregulation of NLRP3 (fig 3A) and upregulation of pro-IL-1B (Fig 3B. 4E. and others; assuming the antibody used for flow is not specific to cleaved, mature IL-1B). Then, caspase-1 cleavage is indicative of inflammasome triggering.
It is unclear how the stimuli and inhibitors used in these studies can achieve both of these endpoints. For instance, MCC950 should reduce cleaved caspase-1 (as it does here), but why does caspase-1 inhibition with ac-YVAD-cmk down regulate IL-1B? Upregulation of pro-IL-1B is upstream of caspase-1 activation. This needs to be reconciled.

4. Regarding HMGB1 data - is a source of HMGB1 known or can it be speculated? Again, HMGB1, as a TLR4 agonist, is a strong primer of NLRP3 inflammasomes. But by itself is not a trigger of inflammasomes. Upregulation of NLRP3, as in point 3 above, needs to be separated from caspase-1 activation. IL-1B cleavage (a downstream event from NLRP3) or secretion (also downstream) are never assessed in the manuscript. I would suggest that assessment of plasma IL-1B levels, or at least immunoblotting to show cleavage of pro-IL-1B, would greatly enhance the ability of the authors to distinguish these two events.

5. Pyroptosis is mentioned in the discussion and is important here. Recent work (Volchuk et al Nature Communications) has shown that in the absence of pyroptosis, inflammasome activation itself does not lead to HMGB1 release. The authors state that since platelets do not have nuclei, that pyroptosis cannot be assessed, but this is not true. As they mention, a combination of caspase-1 cleavage and PI uptake will suffice. PI uptake occurs through gasdermin D in the plasma membrane, and/or through large PM holes induced by lytic death. This does not require a nucleus, and can be assessed in platelets.
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: HMGB1-ACTIVATIED NLRP3 INFLAMMASOME INDUCES THROMBOCYTOPENIA IN HEATSTROKE RAT
Review round: 2
Reviewer: 1

Output:
Basic reporting: My issues have been addressed.
Experimental design: No issues.
Validity of the findings: My concerns on initial review have been addressed.
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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This review article is well written. I suggest some minor changes to improve it.
It is better to include paragraph about measures to reduce contamination of food with mycotoxins. Also, paragraph about the proper care of domestic animals after exposure to mycotoxins.
In figures; there should be figure legend at which all abbreviations used in the figure are written in full term.


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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 1
Reviewer: 2

Output:
Basic reporting: This review article suffers from a number of major flaws in some of the areas covered that calls into question the accuracy with which other aspects of the pathobiology of aflatoxin are appropriately summarized from the literature. There are over 1300 review articles on aflatoxin and its toxicology and this particular manuscript does provide a broader perspective on this compounds impact on a number of organ systems other than liver. However, there are examples of completely incorrect statements from the literature such as on lines 118 and 119 that purports that some of the aflatoxin metabolites can be converted in the sulfates. This is absolutely incorrect with respect to aflatoxin M1 and with respect to aflatoxin Q1 and aflatoxin P1 these are excreted as glucuronides. Further, the citation of the article online 699 with respect to p53 mutations is not appropriate since this is not the first report of these aflatoxin specific mutations that were initially published in Nature in 1991. These types of errors reflect poorly upon the summarization of other data in the submitted manuscript.
Experimental design: As stated above, the final selection of the nearly 120 papers summarized in this review article appears to be biased and not illustrating a curating process to surface the best examples of a given area. Since there are so many reviews available these authors might wish to focus on the most current literature over the past 10 years rather than trying to summarize the nearly 6 decades of work on aflatoxin.
Validity of the findings: The several examples already stated that are fundamentally incorrect in this review article calls into question the care and accuracy with which other literature is summarized.
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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 1
Reviewer: 3

Output:
Basic reporting: The document aims fit the scope of the journal and the topic is still of interest. To my knowledge there is no review that compiles all the data that authors tried to summarize in the present paper. However, I believe that although the paper is interesting, lot of work must be done before publishing it. The paper needs in general a revision of English and technical vocabulary, particularly in the introduction and discussion. Introduction and conclusion are poorly developped.

Lines 56-61: writing should be improved by using technical language such as organoleptic properties.

Lines 86-87: I hardly recommend rephrasing this phase, authors are doing this review based on studies.
Lines 107-148: this part is clear; I suggest specifying in which organisms this pathway occurs.
Line 330: redundant to say male cockerels. The authors may just keep taking about male chickens to avoid confusion.
Line 346: pregnant mothers is commonly use for human beings only
Lines 432-435: should be checked.
Experimental design: The current phrasing makes comprehension difficult, some affirmations are not completely true, some of the references should be changed, the parts in which results from different studies are shown should be double checked to be sure of proper paraphrasing (some examples provided below).

Line 47: AFT are produced by several species, the most important and overspread are those cited in the article, yet they are not the only ones.
Line 49: species that produce AFT are more frequent in the tropics and subtropics, so this phrase must be rephrased as it is not true.
Lines 62-64: Mold produce 4 AFT (AFB1, AFB2, AFG1, AFG2), so these are not derivates.
Line 45: Report of 25% proper citation FAO 1999
Line 68: the reference of AFB1 as powerful carcinogenic belongs to the reports from IARC, EFSA, JECFA since the 1999, so I strongly suggest using one of them as the proper reference, specially IARC and EFSA
Line 82: a citation is missing
Line 711: Modifier Hyglene par Hygiene

Figure and tables:Figure should be worked, as present now they are not clear and the aim of figures are not so clear.  The lack of legends makes difficult to understand the aim of the figures.

Figures:
Figure 1: To my knowledge AFM1, AFP1 and AFQ1 are not completely excreted after been transformed. AFM1 is known to have some toxic effects in mammals and in some to be bioaccumulated.
Figure 2: it is not clear to me the aim of the authors with it, nor why chickens are presented twice.
Figure 3: I do not get why it is shown a maize corn and the fungi on the top, why both?
Tables:
The tables are quite messy. I suggest to organized them, taking into account the information of the columns.  For the species column it should be organized for species, at the moment is a mix of species, gender and age (example for chickens, they are presented as chicks, broiler, cockerels). The doses can also be shown as intervals to make to table more useful. For me it is not clear the solvent column, Does it refer to the extraction solvent?
Validity of the findings: Conclusion is weak and do not show all the work performed through the text.
The arguments are not developped and do not set the expectations.
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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 1
Reviewer: 4

Output:
Basic reporting: Overall the information presented represents valuable information regarding toxic effects of AFB1 in humans and animals along with its toxicokinetic properties. The review is well written, treats an actual problem and concerned on it with details (complexity of AFB1 toxicity) and provides reliable references for future studies. But, I have some comments please address it.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: 1.	Line 33: gives should be giving
2.	Line 43: Productions not produce
3.	Line 47: Aspergillus flavus, A. parasiticus, should be italic
4.	Line 50: 80-90% not 80%-90%
5.	Line66: the citation ( there is a missing reference in the entire text)
6.	Line 81: add and before the last reference
7.	Line 82: citation (a missing reference)
8.	This reference is missing inside the text
Gartung A, Wang Y, Bielenberg DR, Huang S, Kieran MW, Hammock BD, and Panigrahy D. 507 2020. Resolution of eicosanoid/cytokine storm prevents carcinogen and inflammation-initiated 508 hepatocellular cancer progression. Proc Natl Acad Sci U S A 117:21576-21587. 509 10.1073/pnas.2007412117
9.	The references are not arranged alphabetically (line 506 and 510 and so on…..)
10.	 All figures missed caption, it is better to write the full name of the abbreviations under the figures to be more descriptive and understood without having to refer to the main text.
11.	In table (1): in immune-toxicity, 50% ethanol is not used in vivo for mice as a solvent; mice are more sensitive to ethanol-induced lethality. Please revise it again.


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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised manuscript has addressed all of the major critiques in my initial review.
Experimental design: All prior concerns have been adequately addressed.
Validity of the findings: The latest manuscript has updated references and been more inclusive of prior literature.
Additional comments: There are still several minor typographical errors, e.g. lines 397 and 661, that a final editing should 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: RESEARCH PROGRESS IN TOXICOLOGICAL EFFECTS AND MECHANISM OF AFLATOXIN B1 TOXIN
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors edited all the comments and responded to all the comments in details so I recommend accepting the manuscript
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No


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

Input:
Title: ASSISTED MIGRATION AND THE RARE ENDEMIC PLANT SPECIES: THE CASE OF TWO ENDANGERED MEXICAN SPRUCES
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript “ Assisted migration and the rare endemic plant species: The case of two endangered Mexican spruces” is well organized and well written.
The authors have used demographic censuses, field surveys in the search for new stands, and
developed distribution models to achieve the following aims: 1) (i) the exploration of potential new stands and performance of complete demographic censuses to determine individuals numbers, size, and extent of all populations; (ii) construction of models describing the past and current distribution of these species to gain some insight about their distribution dynamics over time; and (iii) identification of areas outside their current ranges (inside Mexico and worldwide) with a high probability of harboring suitable habitats for future assisted migration, by modelling future distributions.
I have some specific comments on this manuscript. Please, see below.
Keywords: they are not keywords that highlight the work
Line 125: What were the factors that explain the extinction? Can you identify them?
Lines 480-496 : this part is a bit confusing, please reduce and make it clearer
Line 470: Our position is that those zones.. Please motivate more strongly
497-499: You affirm that “finally, a parallel strategy of in situ conservation should not be discarded" and In the Conclusion section affirm that " Meanwhile, in situ conservation should be prioritized (Line 519). Although you highlight the point as fundamental in the conclusions, neither you do not emphasize it enough either in the introductory part or in the discussions. Considering that according to Article 8 of the GSPC, in situ conservation is generally considered the primary approach for species conservation as it ensures that species are maintained in their natural environments while ex situ conservation plays a complementary role in situ conservation, providing a “safety back -up ”and an insurance policy against extinction in the wild, can you better rephrase this whole part?
Line 501: You need a stronger conclusion here

Please, add some other references to support the MS:
DOI : https://doi.org/10.1556/168.2018.19.3.3
DOI : doi.org/10.1007/s11258-016-0589-6.
DOI 10.1007/s10531-019-01757-0
DOI : http://dx.doi.org/10.1016/j.foreco.2017.06.044.


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: ASSISTED MIGRATION AND THE RARE ENDEMIC PLANT SPECIES: THE CASE OF TWO ENDANGERED MEXICAN SPRUCES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Abstract.
The beginning of this phrase is confusing: In the coming climatic scenarios, assisted migration would play an important role in the conservation of the threatened plant species… What do you mean by “coming scenarios''? Are you referring to the CMIP6? I saw you used CMIP5. Or do you refer to global warming?

In the background, you also mention “... distribution dynamics”, but I didn’t find any related to this.

Introduction
This section is well organized, but there are points that need to be strengthened and complemented, see above.

Please include in the introduction the non-positive implications of Rescue assisted migration, outside species range, and the country to which it is endemic.
SDM are highly susceptible to producing different results in their geographic projections, future suitable areas for the species will depend on the used method (see https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1365-2699.2006.01460.x). This is a crucial point in your assessment that you didn't consider  thus it reduces its robustness.

Line 133-135. Because you are modeling a only climate scenario, habitat is not a word that comprehensively describes the geographic hypothesis (SDM)
Experimental design: Methods

There are several sections of the methods that are missing or ambiguous.
Lines 153-158. For clarity, please include here how many presence and absence records were used to calibrate RF. How did you consider spatial autocorrelation of species occurrence data? Additional, for RF, Barbet-Massin https://besjournals.onlinelibrary.wiley.com/doi/full/10.1111/j.2041-210X.2011.00172.x recommend: “using the same number of pseudo-absences (or absences) as available presences (averaging several runs if few pseudo-absences) for classification techniques such as boosted regression trees, classification trees and random forest”, but as I mentioned before I do not understand how many presence were used.

Line 185. Specify did you aggregate the cell grid to match 30 arc sec. What did you use, mean, median...?
Lines 196-200. I understand that with this method you select a subset of variables from the initial 42 environmental variables. Please specify which were variables used for each species, in the regional and global analysis. Also, it seems that you didn't reduce collinearity between variables, which is an important part of the modeling process mainly if the aim is to understand which variable limits species distribution.
Lines 200-203. This part is confusing. As I understand it VIM values are just a measure of the contribution of the variables to the model, please correct me if I am wrong. So I don't get how it is possible to normalize VIM values. Or did you normalize the environmental variables?
Lines 205-220. Everything seems OK here, but till the number of unique (unique in a grid-cell of 1 km2 resolution) occurrences (presence and absences) are clear it is very hard to be sure. For the number of stands it seems you used very few records.

Section 2.3

This section needs to be improved. Past and future GCM represents very different possible climate conditions, thus they are a high source of uncertainty. There have been several recommendations to incorporate or show climate storytellings, https://besjournals.onlinelibrary.wiley.com/doi/abs/10.1111/2041-210X.13360.  It is not clear which and how many GCM you used.
Validity of the findings: Please considered all the observations related to your methods, this must be acknowledged to improve the validity of your findings
Additional comments: none


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

Input:
Title: ASSISTED MIGRATION AND THE RARE ENDEMIC PLANT SPECIES: THE CASE OF TWO ENDANGERED MEXICAN SPRUCES
Review round: 2
Reviewer: 1

Output:
Basic reporting: This manuscript now is clearer. The overall idea is very interesting.
Experimental design: The methodology is well described.
Validity of the findings: The manuscript, in general, is clear and well-structured as also the interpretation and the discussion of the results.
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: ASSISTED MIGRATION AND THE RARE ENDEMIC PLANT SPECIES: THE CASE OF TWO ENDANGERED MEXICAN SPRUCES
Review round: 2
Reviewer: 2

Output:
Basic reporting: All comments by the reviewers have been answered appropriately. Results are nicely discussed by the authors, please only adrees my additional comment.
Experimental design: I have nothing to add to this version.
Validity of the findings: Please see additional comments.
Additional comments: You give too much weight to the performance of your model in the discussion, however, you miss to discuss that the high values of for example the AUC are a product of the calibration area. High AUC values can be obtained when calibration áreas are larger and further from presence records.
See https://esajournals.onlinelibrary.wiley.com/doi/10.1890/11-0826.1 and Jorge Lobo's et al. famous paper https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1466-8238.2007.00358.x?casa_token=edB-w_hRSJQAAAAA:_T8sGSISwou-ZrcZBnuDfKDmQFl6aEIQMe9wGy2p4NlD74xgK5ZDnbXC8HKQdEpe5xrYuvVT4lQPMxLo


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: CRITERIA FOR ASSESSING THE ADAPTIVE CHANGES IN MIXED MARTIAL ARTS (MMA) ATHLETES OF STRIKE FIGHTING STYLE IN DIFFERENT TRAINING LOAD REGIMES
Review round: 1
Reviewer: 1

Output:
Basic reporting: - The English language is at a satisfactory level throughout the manuscript.

- The criterion is partially fulfilled.

- The article lacks simplicity and clarity. On the other hand, readers will have no trouble understanding tables and figure.

- The obtained results are in accordance with the research hypothesis.
Experimental design: - The presented study is in line with the aims and scope of the journal.

- The criterion is fulfilled. The findings of this research are innovative and represent a significant progress in the field of training load design in combat sports athletes.

- The criterion is partially fulfilled.

- The methods section provided enough information for readers.
Validity of the findings: - The criterion is fulfilled.

- Fundamental data are unambiguous and are clearly presented to the readership.

- The conclusion is the weakest part of the entire study. The authors must once again check this section.
Additional comments: •	The abstract is really extensive and contains too many words. The results within the mentioned section have several unnecessary claims.
•	Lines 75-78: Can you provide references for this statement, please? Are there researchers who have previously applied listed indicators to assess the effectiveness of different training loads in athletes?
•	The term "various/other sports" is frequent used in manuscript. It would be clearer for the readers if you would report more precisely which sports do you mean? Combat sports or?
•	Lines 113-115 and 145-148: Do you mean these assertions are indispensable? Please, remove them from the text.
•	Authors should change term "Methods" to "Measurements". In order to improve the readability and clarity of the text, it is also recommended to add subtitles for each separate measurements (maximal muscle strength, special training, body composition and biochemical parameters).
•	Why did you explain the findings in the results section? This is more appropriate for discussion. Check all paragraphs again, please!
•	Line 231: How can researchers be sure that the results obtained are occur due to the increased levels of intramuscular and intermuscular coordination of athletes? Are there any previous findings/references that can confirm this hypothesis? Additionally, this statement is also more appropriate for discussion.
•	Line 240: The abbreviations in Table 4 should be placed below it.
•	Please improve the visibility of Figures 1 and 2. Check the text above the figures again, please.
•	Line 287: "The obtained results" – The mentioned terms are often used in the results section, which will be boring for the readership.
•	Line 298: Please report results for group A that implemented a medium intensity training load.
•	The discussion requires certain corrections. The authors analyzed only biochemical parameters, and what about other measured indicators?
•	Lines 330-332: The mentioned contradictory findings need to be explained.
•	Lines 360-364: This statement should be moved to discussion.
•	Lines 365-380: You just repeated the results obtained. Please, remove this text from the conclusion section.


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: CRITERIA FOR ASSESSING THE ADAPTIVE CHANGES IN MIXED MARTIAL ARTS (MMA) ATHLETES OF STRIKE FIGHTING STYLE IN DIFFERENT TRAINING LOAD REGIMES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The evaluated paper investigates the adaptive reactions of young athletes (MMA fighters) to 2 regimes of effort (medium and high intensity and load). The value of the study results from the high number of measured and analyzed parameters (data related to the accuracy of the movements execution technique, body composition, biochemical indicators of blood and analysis of the associations between these independent, studied variables). Laboratory investigations are the strong point of this study. The research is useful for the scientific direction of the training process for the targeted athlete category.

The reviewed work is valuable and useful to those working in the field of sports (especially due to detailed laboratory explorations) and may be published after a few minor revisions.
Experimental design: I think it would be useful to formulate working hypotheses (at the end of the introduction), to which the results from the tables and graphs should be reported.

Please specify the (MMA) sports activity experience of the 2 groups studied and some data related to the height and weight of these athletes (average and standard deviation).

Could you indicate the 2-month interval in which you performed the experiment (months and year)?

In what stage of training (general physical training, pre-competitive or competitive training) were the investigated fighters?

Please specify the type of study performed (longitudinal or cross-sectional study).
Validity of the findings: Why didn't you compare the results between the 2 groups (A and B as independent samples) by the non-parametric Mann-Whitney U test? I noticed that you have a large volume of analyzed data anyway (and these additional comparisons would have doubled the number of tables), but it would be advisable to make these comparisons perhaps in another paper, for a complete view of your results. However, this does not reduce the value of the study.

Tables 2-5 show the results of the Friedman-Anova test and compare the differences between the 3 tests (T1 - initial, T2 - intermediate - one month and T3 - final - 2 months). It should result in 3 data pairs with 3 values of Z and the associated significance thresholds: T1-T2, T1-T3 and T2-T3. Only two pairs appear in all tables, probably T1-T2 and T2-T3. Could you include the missing pair?

You used SPSS software, so the effect size value (expressed by Kendall W) might have been calculated if this option was checked. It would provide additional support in arguing for the significance of the differences obtained. It's just a remark, the study is well-argued enough even without this statistical indicator.

In Table 4, for the BF indicator (body fat %) there are big differences between groups A and B, right from the initial testing. What is the explanation?

Figure1 and 2- The title: Correlation between biochemical parameters of blood in group A athletes before (A and C) and after training load (B and D) during 2 months of research in the conditions of medium intensity training load regime (Ra = 0.64). Below figures, there appear in the initial tests A and B, respectively in the final ones (after 2 months) C and D.
Additional comments: You refer to the morpho-functional parameters (line 22). This wording / expression is repeated in "Participants" (line 114), respectively in "Conclusions" (line 359). Anthropometric parameters (height, body weight, perimeters and diameters etc. are not presented in this study).

You could include a short paragraph to identify issues related to the specificity of physical effort in MMA (which differentiates it from other combat sports).

Line 245:  Table 5 presents the results of changes in biochemical inadicators of the blood serum…. I think the word indicators is 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: CRITERIA FOR ASSESSING THE ADAPTIVE CHANGES IN MIXED MARTIAL ARTS (MMA) ATHLETES OF STRIKE FIGHTING STYLE IN DIFFERENT TRAINING LOAD REGIMES
Review round: 3
Reviewer: 1

Output:
Basic reporting: The authors fulfilled all criteria relating to the "Basic reporting" section.
Experimental design: The authors successfully responded to comments associated with the "Experimental design" section.
Validity of the findings: The authors fulfilled all criteria relating to the "Validity of the findings" section.
Additional comments: I have 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: CRITERIA FOR ASSESSING THE ADAPTIVE CHANGES IN MIXED MARTIAL ARTS (MMA) ATHLETES OF STRIKE FIGHTING STYLE IN DIFFERENT TRAINING LOAD REGIMES
Review round: 3
Reviewer: 2

Output:
Basic reporting: I believe that the explanations mentioned by the authors and the changes made in this scientific article would allow publication in this version.
Experimental design: The authors made most of the suggested changes, or provided reasoned explanations, when they had a different approach to the issues previously reported.
Validity of the findings: The authors made most of the suggested changes, or provided reasoned explanations, when they had a different approach to the issues previously reported.
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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: The authors investigated few databases. It is mandatory to investigate Web of Science and EMBASE.

It is also mandatory to investigate the gray literature.

The authors do not assess the certainty of evidence with GRADE. It is currently considered a mandatory and necessary step in systematic reviews with or without meta-analysis.
Validity of the findings: The few databases investigated leave doubts as to whether all the articles were retrieved in the search.

Without GRADE, doubts persist as to the certainty of evidence of the selected articles.
Additional comments: It is necessary to review the search strategy, investigate other databases and gray literature, and assess the certainty of evidence by GRADE.


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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: -	The spelling and language overall are not great. You put capital letters where it is not needed, you make grammatical errors, you use brackets but never close the bracket, …I suggest that you check your manuscript for these little errors and let someone who is proficient in English review the manuscript.

Abstract:
-	Your abstract is very dense and detailed, the aim and the most important aspects of this review are not clear.

Introduction:
-	There is no need for figure 1 or 2, they do not add anything to your manuscript.
-	Too much information that is not relevant for the reader: the most prevalent cancers with their numbers don’t need to be described. You can speak on this as a more general description, but this is too detailed and not needed.
-	Reference 9: you use this reference when you talk about patients in general, however, this reference is of a study on prostate cancer. This reference is not relevant. This is a general remark: use references that are relevant to your review fe. all cancer patients, as your review is also on general cancer patients.
-	In your introduction, it should be clear what you mean when you talk about cancer patients. Is this anyone who has a diagnosis or is in active treatment, those who are cancer survivors?
-	Line 90-92: So QoL is not determined by cardiovascular fitness or exercise capacity but only by sleep quality and feeling empowered?
-	The evidence of exercise and physical activity on cancer-related symptoms is vast, I want to see better references to this. Even on recurrence rate, survival rate, … The importance of exercise is not really clear from your introduction
-	Line 112-115: So you will only look at cancer patients suffering from cancer-related tiredness?
-	The aim of your review remains very unclear after your introduction
-	Your introduction right now is just a summation of facts, I miss a comprehensive introduction. Use the problem-gap-hook method.

Figures & Tables
-	Figures 1 & 2 are not necessary
-	Table 4: you report on different outcomes than the ones you are interested in: fatigue, anxiety, sleep, strength, …
Experimental design: What is your actual research question? You mention cancer patients suffering from cancer-related tiredness, but then you don't mention this anywhere else?

Methods:
-	I (Intervention) = Exercise and QOL : how is QoL an intervention?
-	Why do you include these specific journals, are they not available through pubmed, scopus search?
-	Line 155-156: Why only between 2010-2020 ?
-	Line 182-183: If two reviewers could agree? How did you agree? Did you not use the scoring results as a guide on whether to in or exclude the study based on methodological quality?
-	In and exclusion criteria are very broad for example: What are cancer patients? is this anyone with a diagnosis, anyone with curative treatment, those with a terminal diagnosis, those who survived cancer and are post-treatment,… ?
If you include all these type of patients in your review, you have too much heterogeneity
-	High-intensity exercise is an exclusion criteria, why?

Results:
-	Line 187-190: You retrieved another 287 articles by hand searching? This is an indication that your search strategy was not good and that in fact you haven’t really conducted a proper systematic review
-	Line 207: if you only looked at studies between 2010-2020 then it is obvious you will only find those articles.
-	You describe in detail the journals from which you retrieved the articles (line 197-207) but you do not do these when it comes to the actual relevant information. I want to know more about the content and not which journal it was published in. What type of exercise has an influence on what type of immunity markers, what type of patients benefit of this exercise? Make a synthesis of your findings in the result section.
Validity of the findings: You results: you only describe these results in table 4 but do you have the data on this (p-values, ...)?

Discussion:
Your discussion has no common thread. You might check some of prisma statements but I miss an in-depth discussion. I do not find any discussion on your own findings, potential mechanisms why exercise could have these results, recommendations, critical remarks, …

-	Line 253-260: It was found that IL-8 and neutrophil-activating protein-78 levels in the home-exercise group were substantially lower after the exercise-programme --> but not in the supervised exercise group: how do you explain this?
-	line 292-294: I don’t understand the relevance of this sentence, what do you mean? Is this a recommendation for future research, what medication?
Additional comments: I have read the manuscript with great interest on this very relevant topic.
However, this review lacks quality. My most important remark is that you incorporate a lot of aspects but are never really to the point. Therefore the review is often incomprehensive. The description of the methods used is insufficient, the reporting of the results is unclear and the discussion section is not strong and does not add to the manuscript. This manuscript would benefit from an extensive revision before being considered further for publication.
I composed my more detailed remarks in the sections and gave some suggestions on how you can improve this 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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 3

Output:
Basic reporting: General comments
The study carries an appreciable novelty, considering the continuously increasing incidences of cancer on a global scale. The authors have ingeniously considered reviewing the effect of physical activity as a panacea to the cancer issue. However, there are several concerns with the manuscript, and a major revision is needful.

Technical arrangement and accuracy
The construction of several sections of the manuscript should be reordered for soundness and technical comprehension. For example, Lines 35-36 reads “ A formal search of the keywords was performed before proceeding with THE REGISTRATION AND FURTHER for the systematic review in order to make sure the availability of enough…” This statement, though somewhat valid, sounds quite twisted and might be difficult to comprehend. A revision might improve it.
Authors stated in lines 61-62 “Cervical cancer is one of the most prevalent cancers in females…”  Then, in the latter parts, precisely lines 69-70, it was stated that “On the other hand, Breast carcinoma is REGARDED AS A THE MOST frequent medical illness that women face across the world”. The statement in lines 69-70 is both grammatically incorrect and inconsistent to the fact stated in lines 61-62. Furthermore, if other cancers peculiar to women would be mentioned in the introduction, it might be suggestable for them to follow one path. They shouldn’t be mentioned in a paragraph in lines 61-62 and repeated without any different point in lines 69. They should be ordered in one place in the introduction section.
References were made to GCO (the Global Cancer Observatory (fmr GLOBOCAN)) in lines 62-63, and the citation was not offered. Though the citation was apparently offered in the next sentence (line 65), but it would technically suffice if the source that mentioned GCO was mentioned in line 63
In line 73, it was mentioned that “New Zealand than in INTERMEDIATE ADVANCED NATIONS such as Europe (central)…” There is no technical classification of any set of nations as intermediate-advanced, if there is, the authors can provide a reference for this. The conventional classification, according to world bank, is based on Gross National Income per capita as low income countries (similar to least developed countries), lower-middle income countries, upper-middle income countries, high income countries (similar to developed countries) (https://blogs.worldbank.org/opendata/new-country-classifications-income-level-2019-2020). This should be revised.
In line 79-80, it was stated that “Physical activities can be explained as skeletal, muscular ambulation or work that leads in energy dissipation”. An explanation as this should have a recent and citable reference literature.
Authors declared in lines 103-104 that “Cancer incidence and immune system dysfunction ARE ALSO age-related changes that can be mitigated to some extent by exercise”. This claim might be contentious, as young people in some populations have been diagnosed with cancer and immune issues. “ARE ALSO” can be suggestively substituted with “can also be as a result of…”
In line 108, the word EXERCISE was capitalized. This should be reviewed if such punctuation has no significance to the context.
Study rationale
This section is seemingly wrongly positioned and of little technicality. It looks like a declaration of results “We found that…” It is, as other several parts of the manuscript, replete with the use of the pronoun WE which should not be used for technical writing.
Line 119 states “effects. we found that Natural Killer cytotoxic activity increased after…” This line has issues related to 1. wrong capitalization for the commencement of a fresh sentence 2. The use of the pronoun “we” 3. An undefined term “Natural Killer” introduced with questionable capitalization 4. “Natural Killer cytotoxic activity” which seems to be vague and difficult to understand.
The entire rationale should be part of the introduction and should not be stay distinct. It should be short, very technical, and should be an executive justification for why the study was conducted, and the problem hoped to be solved with some novel approach.
Experimental design: Materials and methods
No explanation was given for why the criteria were selected for eligibility, studies, data extraction, and quality assessment regions. For example, why was the Joanna Briggs Institute (JBI) critical assessment checklist used, of all checklists? Is it the only checklist that can be used? What is its advantage? Why were only 2 authors and 2 reviewers used? Why not more? Why not less? The questions should be answered in concise but technical terms in each section
Line 162 states “Two of the reviewers (S.B and H.A) WERE PERFORMED THE INCLUSION…” This grammar should be checked for soundness.
Validity of the findings: Discussion
Lines 225-250 are recommendable only for the justification aspect of the introduction. They do not discuss results obtained.
Excessive use of the expression “the current systematic review” could make the discussion monotonous.
The question of HOW? was sparingly answered all through the discussion. For example, the Tumour Necrosis Factor alpha mentioned in lines 236-237 was not explained in full detail regarding how it, and IL-6 are reduced by exercise. A brief but comprehensive biochemical mechanism of HOW IT HAPPENS between the input and output variables should be offered. This would make the discussion full of sound information, and not just a comparative declaration with past studies.

Limitation and Conclusion.
The use of WE should be completely removed and revised. For example, “We included published original articles…” should be revised to “Published original articles were included…”
Authors mentioned in lines 311-312 that “Understanding the mechanism between exercise and immune function in various cancer patients, as well as the relationship between these immunological markers and clinical outcomes, ARE ESSENTIAL FOR THE FUTURE STUDIES.” A basic search from google scholar and other scientific literature sources would show that several PAST studies have defined tis relationship to some extent. It is suggested that the relationships already published be mentioned in the discussion section.
Additional comments: The grammatical construction of the entire manuscript should be revised. For example, the excessive and unwarranted use of semi-colon ";".


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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 2
Reviewer: 1

Output:
Basic reporting: I see a lot of improvement in this area.
Some suggestions:

Line 26: cancer does not only affect the body, but also has a psychological, societal and economic impact.
Line 27: The physical activity --> Physical activity (no need for The)

Line 129:
Inclusion criteria
- You should add that you only include studies that have a physical activity/exercise intervention and those who look at your outcome criteria. The inclusion criteria you actually mentioned in line 123-124. As of now, your list is not complete.

Line 201-208: For me, it would make more sense in you report on this before the actual findings from the studies (line 181-200).

Line 256: exercise-programs and not Programs

Table 2:
Your results and conclusion column is very similar. Maybe in the results column, you can work schematically and less with text and sentences. So it is very clear what your findings were and what findings were significant

 Supplementary file 1 table 2:
Could you add the questions of the JBI in the legend of this table. Then the table can be read separately from the manuscript.
Experimental design: The research aim is now clear.

Methods:
Line 114:
You incorporated another database Sciencedirect on 15-02-2022. However, did you repeat your search strategy again in PubMed, Scopus and Cochrane? A check of these databases might show more relevant literature.

Results:
Overall, you could make a synthesis based on your three outcome categories: QoL, fatigue and immunity markers. And describe your results per study in those categories. Your table 2 gives an overview of each study, so within your result section, I want to find a synthesis and not just the same overview in text.
Also, were the outcome measures only measured once? What about follow-up? Was this reported in the studies, it might be relevant to know how these outcome measures change with time.
Validity of the findings: Discussion:
Line 224-226: You report on the findings on an increase in NK-cell cytotoxicity, could you elaborate on that. What does this mean for a cancer patient in treatment?
Line 224-227: You report on the findings on NK-cell cytotoxicity, but do not mention anything of your other outcome measures. Could you add very briefly what the most important findings were on all predefined outcome measures (Qol, fatigue, immunity markers) and then proceed with the rest of the discussion, where you go in depth on each outcome. Non-significant findings are also important findings.

Overall, I do miss a mention of limitations with your own review. The fact that you have such heterogenous studies (all types of cancer patients, all stages, all types of treatment, all ages, all types of physical activity …).  Do you see any explanation why some studies did not find significant results? Maybe you can see that some types of physical activity seem to work better then others. This all stays quite general. You do mention that in future research, studies are needed that investigate the different forms of exercise. But perhaps, you already see some aspects emerge from your data.

Conclusion
This is very general. What does this systematic review specifically contribute to this topic?
Additional comments: I see many improvements in the basic reporting and methods area. However, a more in-depth discussion section will add a lot of value to this systematic 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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revised manuscript is a greatly improved version of the previous one. The authors have shown mastery of the research procedure involved in the study and have made important rectifications. However, there are some minor suggestions that might further improve the scholarly appeal of the manuscript.
Experimental design: This section is satisfactory.
Validity of the findings: The conclusion for such an important study is quite short and nominal. The declaration that "Regular exercise has been shown to improve QOL, reduce depression and improve immunity
markers in cancer patients" is a quite generall known fact. The same principle is typically advised for a normal healthy lifestyle.

The question is, what is the unique finding and coclusion of the study? How are those unique findings going to affect the future? What steps should stakeholders (patients and health personnels) take to achieve better outcomes for cancer management?

These thoughts can be addressed in the conclusion section.
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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 3
Reviewer: 1

Output:
Basic reporting: Still some issues with spelling or grammatical errors. So I would suggest to let a fluent English speaker read your work as a whole again.
Some examples:
Line 53: prescribe? You mean describe?
Line 226 these findings suggested -->  suggest that: present tense is more suited
Line 295: a rehabilitation programs --> program : singular in stead of plural
Experimental design: No comment
Validity of the findings: Line 46-48 : Living an active lifestyle before diagnosis. Before diagnosis, cancer patients are healthy people. Can you make your recommendation on cancer patients while undergoing treatment or after treatment?

Line 332: Why don't you mention the heterogeneity of the included studies as a limitation and strength? Limitation as it is difficult to say anything about one specific type of cancer, but you give a broad overview of the cancer population as a whole.
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: EFFECTIVENESS OF PHYSICAL ACTIVITY ON IMMUNITY MARKERS AND QUALITY OF LIFE IN CANCER PATIENT: A SYSTEMATIC REVIEW
Review round: 3
Reviewer: 2

Output:
Basic reporting: The revised manuscript has been greatly improved
Experimental design: This section is satisfactory.
Validity of the findings: This section is satisfactory.
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: PREVALENCE OF SARCOPENIA WAS HIGHER IN WOMEN THAN IN MEN: A CROSS-SECTIONAL STUDY FROM A RURAL AREA IN EASTERN CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please see general comments
Experimental design: Please see general comments
Validity of the findings: Please see general comments
Additional comments: This manuscript is a straight forward descriptive study of the prevalence of sarcopenia in a rural area in China, documenting that the prevalence in women is higher than in men, even after adjusting for co-variates. This is a finding that is contrary to the majority of studies.

The manuscript could be improved in the following ways:
1)	Literature review: please include references for gender differences in urban settings, and among non-Chinese populations. In particular please include prevalence studies in urban settings in China: for example there have been many studies on this topic from the West China Medical University, directed by Prof Dong Birong.
2)	Please show the response rate of all the eligible population, and reason(s) for non-response, so that any potential bias may be detected.
3)	Please show the prevalence of sarcopenia for men and women, before and after adjustment for confounding factors.
4)	Please show the factors associated with sarcopenia (using variables shown in Table 1) using logistic regression, separately for men and women.


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: PREVALENCE OF SARCOPENIA WAS HIGHER IN WOMEN THAN IN MEN: A CROSS-SECTIONAL STUDY FROM A RURAL AREA IN EASTERN CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The writing of this article is clear in English. It provides adequate background info with relevant and sufficient references. The data structure including figures and tables is appropriate.
Experimental design: This is a national multi-center study in China. The data reported in this paper are cross-sectional with relevant survey info and objective measures. The research question is clearly defined and scientific approach is vigorous and described in detail with no ethical concern.
Validity of the findings: Sex and gender difference in sarcopenia is important with potentially high impact. This article reports underlying data that are robust to address this research question. The conclusion is well stated and supportive of the original hypothesis.
Additional comments: One suggestion for the "Conclusions" section (in both Abstract and at the last section of the text) is to change "eliminate" to "address" in the statement " ...to eliminate such gender disparity in health." as such disparity would likely be completed eliminated.


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: PREVALENCE OF SARCOPENIA WAS HIGHER IN WOMEN THAN IN MEN: A CROSS-SECTIONAL STUDY FROM A RURAL AREA IN EASTERN CHINA
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1) Thank you for providing the raw data, however the data should be in excel format instead of stata format for easy viewing

2) The english can be improved by checking through subject verb agreements e.g. line 31 should be "women in rural areas in China seem more vulnerable…” and line 37 “decline in quality of life".
Experimental design: No comment
Validity of the findings: No comment
Additional comments: 1) In the abstract section under results, “the prevalence of sarcopenia was 151% times higher…” this sentence be rephrased as proportion of people with sarcopenia is 1.5-fold greater... (95%CI 1.01-2.26, P = xxx), with the actual p value stated. Please do the same for the results section lines 139-142.

2) Lines 40-41 seems to be linked to 36-37. Suggest to rephrase into 1 sentence.

3) Lines 98-100 - it is unclear why the authors divided multimorbidity into 4 groups rather than presence/absence of multimorbidity or number of of chronic diseases as a continuous variable.

4) Lines 102 - While the types and time per week for physical activity was determined, this was not reflected in Table 1 where the physical activity variable was binary.

5) Lines 117-118 - “Two-sample t-test…mean age were 72.4(0.31) years…” Please check and rephrase this sentence as t-test should not be needed for describing the mean age, provided there is a difference between groups, which in that case, the p values should be provided. Please also describe what the numbers in the brackets mean e.g. SD?

6) In Table 3, please state what each model represents for easy reference at the footnote

7) The definition of physical exercise is vague in the study - does this include non-recreational physical activity such as transportation, occupational, housework? Non-recreational physical activity has been shown to be higher in less developed areas, compared to developed areas where people engage largely in recreational physical activity (due to transition towards sedentary occupations and motorised vehicles etc.) This should perhaps be included in the discussion.

8) Was socio-economic status measured in this study? If so, it should be included in the descriptive statistics, as the authors mentioned that women are disadvantaged in terms of income levels - which might also affect a myriad of lifestyle factors and hence sarcopenia.


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: PREVALENCE OF SARCOPENIA WAS HIGHER IN WOMEN THAN IN MEN: A CROSS-SECTIONAL STUDY FROM A RURAL AREA IN EASTERN CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: 1. Since income levels (socioeconomic status) differed between men and women, this should be accounted for in the subsequent analyses.

2. Please provide a reference for the adjustment of co-morbidities in a non-linear manner as the authors have done in this paper.

3. In the introduction and discussion sections, it is not necessary to mention the Professor's name as these would be covered in the cited references.

4. The literature gaps need to be identified and justified clearly in the introduction. In the discussion, the authors mentioned Gao et al 2015 has also investigated sarcopenia prevalence in rural western China. How does this present study differ from earlier studies? What new knowledge is added?


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: CLINICAL CHARACTERISTICS OF THE SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 OMICRON VARIANT COMPARED WITH THE DELTA VARIANT: A RETROSPECTIVE CASE-CONTROL STUDY OF 318 OUTPATIENTS FROM A SINGLE SIGHT INSTITUTE IN JAPAN
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: 1. which type of oxygen therapy you mean, is include  CPAP
2. SPO2 measurement is the determinant for need of oxygen therapy or other factor?
3. can you make the sample size more than that
Validity of the findings: 1.why you didn't consider the D dimer and ferritin as prognostic markers?
2. how you can explain AST has significant value and ALT not
3. why the age is significant for oxygen therapy
Additional comments: 1. what about calculating  AUC by taking the combining of more than one test ex: crp+ldh vs crp alone
2.can you explain the non significant  effect of vaccine


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: CLINICAL CHARACTERISTICS OF THE SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 OMICRON VARIANT COMPARED WITH THE DELTA VARIANT: A RETROSPECTIVE CASE-CONTROL STUDY OF 318 OUTPATIENTS FROM A SINGLE SIGHT INSTITUTE IN JAPAN
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this study, the authors to identify the biomarkers predictive of the need for oxygen therapy in patients with COVID-19. The study is interesting, but in the literature there are many published studies.
I suggest doing a more exhaustive review of the literature, including systematic reviews. Consider "Malik et al. Biomarkers and outcomes of COVID-19 hospitalisations: systematic review and meta-analysis. BMJ Evid Based Med. 2021 Jun;26(3):107-108. "

There are grammatical errors. The language can be improved.
Title
There is an error in the title "COVID-2019", it is COVID-19
Title: include the study design following the STROBE guidelines
 Abstract.
Include the design in the Methods section.
Include a conclusion section.
Experimental design: Methods section needs more details

Include the study design following the STROBE guidelines.
Include with supplementary information checklist STROBE.
How was a case defined for inclusion in the study? I suggest detailing the inclusion and exclusion criteria.
The criteria for assigning each group should be detailed in the methods section.
Validity of the findings: Results
I suggest including the STROBE flowchart.
Line 113. We divided the patients into two groups according to the need for oxygen therapy. The criteria for assigning each group should be detailed in the methods section.
Table 1 and lines 120-130. I suggest performing bivariate and multivariate analyzes (crude and adjusted ORs) for all significant results. I suggest correctly interpreting the results "The median patient age was 55 (45.5–72.25) years among those requiring oxygen therapy, versus 44.5 (34.3–52.8) years among those who did not require oxygen therapy (P < 0.001)".

Line 177. There is an error should be figure 4. Alternatively, the analyzes in figure 4 could be performed with adjusted HR instead of OR.

There are two variables that should be included probability of ICU admission and mortality. In addition, it is important to evaluate biomarkers according to the clinical course, i.e., mild, moderate and severe disease. I suggest including these analyzes in the study.

Discussion.
In addition to the limitations described, the authors must include and discuss the sample size and that the two groups may not be comparable (102 vs. 40 patients).

Conclusions.
I suggest revising the conclusions based on the previous comments.
Additional comments: No 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: HYDROLOGICAL AND LOCK OPERATION CONDITIONS ASSOCIATED WITH PADDLEFISH AND BIGHEADED CARP DAM PASSAGE ON A LARGE AND SMALL SCALE IN THE UPPER MISSISSIPPI RIVER (POOLS 14–18)
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this manuscript, the authors present the findings of an acoustic telemetry study tracking movement of paddlefish, silver carp, and bighead carp through the Mississippi River Lock and Dam 14-18.  The field study encompasses a large spatial region and provides insights into fish movement through Mississippi River Lock and Dams that have not been fully described in the past.  While the sample sizes of silver and bighead carp are severely limited, the methodology and statistical analysis follow current best practices.  The authors found that passage of all three species was closely tied to hydraulic head at the lock and dams among other covariates including lock usage, and water temperature.  Overall, the manuscript is generally well written and methods adequately described.
Experimental design: The only major concern / question I have regarding the interpretation of the results is the reliance on hydraulic head as the sole covariate for hydraulic conditions at a lock and dam.  While hydraulic head is an easily quantifiable variable, it is a poor descriptor of localized hydraulics that actually influence fish passage.  For example, the same hydraulic head (difference between head- and tail-water levels) can be maintained by opening all spillway gates to the same, small opening versus opening only a few gates to a larger opening.  Characteristic water velocity and turbulence downstream of each opening is going to vary drastically in spatial scale and impact on fish movement behavior.  The authors need to acknowledge this imprecision in hydraulic head and address how this might limit insights from the current data set.
Validity of the findings: The findings are of general use for decisions regarding control strategies for invasive bigheaded carp, and are thus of interest for the journal audience.
Additional comments: Detailed comments are listed below with corresponding line numbers:
Line 86 – Insert “downstream” before “migration”
Line 93 – “Intentionally fragmented systems” is an awkward wording.  I believe the authors mean to say “barriers” can prevent the spread of nonnative species.
Line 95 – While seasonal barriers are used to control sea lamprey in the Great Lakes, they are by no means “effective” at a broad scale as the migratory period of many native fish overlap substantially with sea lamprey migrations, as described by Velez-Espino et al. (2011).
Line 99 – Add Rahel and McLaughlin, 2018 here.
Line 103 – Delete “main thalweg”.
Line 106 – Replace “current” with “water”, replace “gates” with “spillway, and insert “gate” before “controlled”.
Line 107 – Lock and Dam #1 can’t be in open-river conditions due to the spillway type, and Lock and Dam 5 and 11 are also very similar in open-river occurrence.
Line 108 – Add reference to Wilcox et al. 2004.
Line 138 – Work by Zielinski et al. (2018) and Finger et al. (2019) have expanded on this possibility greatly.
Line 139-140 – This sentence is awkward, consider revision.
Line 188 – Are the same tag tpyes used for paddlefish and silver and bighead carp?
Line 216 – How representative is a 1-m depth to where paddlefish swim?
Line 234 – Why is hydraulic head used in the statistical analysis versus discharge or gate opening (see comment above)?
Line 251-255 – Why aren’t lock usage used in the TTE analysis but is used in later analyses?
Line 357-359 – Was any range testing done with a barge moving through the array?  Presumably, the boats will cause line-of-sight issues and noise that may reduce tag detection.
Line 388 – 397 – What about bigheaded carp?
Line 417-420 – It might be useful for the authors to expand why up- and down-stream tows were analyzed separately.  Is there something about lock operation in either scenario that differs and would impact the ability for fish to pass?
Line 493 – What conditions were present during the upstream passages that occurred outside of open-river?
Line 522 – While velocities through the partially closed gates may be high immediately under the gates, the spatial extent can be limited and create recirculation zones at different depths in the water column that could permit fish to nearly reach the gate without experiencing localized regions of high velocities.  As part of this study, the authors have little to no quantified data on the flow conditions in or near the gates and thus discussion on specific flow conditions influencing fish movement is purely conjecture at this point.  The flow conditions in and around the spillway gates are predictably very complex and citing data like “velocities as low as 0.6 m/s” from Wilcox (1999) simply perpetuates our general lack of understanding on the spatiotemporal hydraulic characteristics.
Line 526 – Morphological differences from bigheaded carp or interindividual differences?  Please be more specific.
Line 524 – The reference to Wilcox (1999) is misleading at best.  The value of “0.6 m/s” was not recorded, but was estimated from a physical model study completed by Markussen & Wilhelms (1987).  The region of flow where 0.6 m/s was also spatially limited and not representative of the broader heterogeneity of the flow field through a spillway gate.
Markussen, J.V., Wilhelms, S.C., 1987. Scour Protection for Lock and Dams 2-10, Upper Mississippi River. Hydraulic Model Investigation. Technical Report HL-87-4. US Army Engineer Waterways Experiment Station.
Line 541 – This statement is counter intuitive as the highest localized velocities are expected to occur during high hydraulic head, which occurs during low discharge.
Line 608 – Large metal objects like the miter gates?
Line 621-626 – These recommendations related to acoustic deterrents are a bit outside the scope of this study and are not necessary.  There are substantial lines of research already directed toward acoustic deterrents that are more current than Lovell et al. (2005 & 2006).
Line 650 – This exact idea has already been proposed by Zielinski and Sorensen (2021).
Zielinski, D. P. and Sorensen, P. W. (2021) Numeric simulation demonstrates that the upstream movement of invasive bigheaded carp can be blocked at sets of Mississippi River locks-and-dams using a combination of optimized spillway gate operations, lock deterrents, and carp removal. Fishes, 6(10), https://doi.org/10.3390/fishes6020010.


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: HYDROLOGICAL AND LOCK OPERATION CONDITIONS ASSOCIATED WITH PADDLEFISH AND BIGHEADED CARP DAM PASSAGE ON A LARGE AND SMALL SCALE IN THE UPPER MISSISSIPPI RIVER (POOLS 14–18)
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1.	Definition of Terms
Lines 56, 67, 68, 86, 103-104, 136, 332, 467, 469, 471, 480, 485, 493, 517.
Comment: The terminology is confusing, 1) the definition of open-river needs improvement, because technically there isn’t a “main thalweg spillway gates” part of the river, and 2) the generic use of “spillway passage” can refer to two very different parts of a dam; the adjustable spillway gates used to control water flow in the main channel (i.e. tainter and roller gates), and the fixed-crest spillway designed to release surplus flood water.
Recommendation:  Change all parts of the document that refer to “spillway passage” to reflect that fish passed through the adjustable spillway gate openings in the main channel when the gates were positioned in the Open River condition.  A generic discussion of terms could be helpful:

•	Open River - occurs when the dam gates are lifted above the water, passing unobstructed flow through the adjustable spillway gates.  During open river the head and tail water surface elevations of the river are nearly equal, essentially following the natural slope of the river.
•	Spillway –Seventeen of the UMR navigation dams have fixed-crest spillways in the earthen dike sections of the dams, which are designed to pass flow during periods of high discharge. Most of these spillways have an ogee crest.  Lock and Dam 15 is the only dam within the study area that does not have a fixed crest spillway.  All water flows though the roller gates at L&D 15 and through the hydropower dams in Sylvan Slough.
•	Thalweg - The thalweg is the deepest cross-sectional area of a river.  At UMR dams the thalweg is technically in either the main lock or the auxiliary lock chambers.  Most UMR dams are designed so that the deepest part of the river allows for navigation traffic and emergency dewatering of the upstream pool.
Experimental design: No comment
Validity of the findings: 2.	Un-cited Literature - Several additional papers should be considered in the discussion

Lines 138-142 and 641-660
Comment:  Gate manipulations have been considered to block fish movements in addition to those mentioned on line 142.  I was expecting to see some discussion/comparisons with the similar fish movement study conducted at L&D 2 (even though the species are different).

Recommendation: If not further discussed, at a minimum, Finger et al (2020) should be referenced and gate manipulations added to the list of deterrents on line 142.

Finger, Jean Sebastien, Andrew Thomas Riesgraf, Daniel Patrick Zielinski, Peter William Sorensen. 2020. Monitoring upstream fish passage through a Mississippi River lock and dam reveals species differences in lock chamber usage and supports a fish passage model which describes velocity‐dependent passage through spillway gates.  River Research and Applications 36:36–46.

Lines 549 – 556
Comment:  Add a discussion regarding Maynord (2005) observations to explain the hydraulic differences between upbound and downbound lockage.

Recommendation:  Add information regarding; “The average upbound unloaded tow from the field study passed 49 percent of the water volume of the lock through the wheels. The average downbound loaded tow from the field study passed 228 percent of the water volume of the lock through the wheels (Maynord 2005).” plus a discussion on how this information may have impacted fish passage in the lock.

Maynord, Stephen T. 2005. Flow through the wheels of towboats in 600 ft locks on the UMR-IWW.  Upper Mississippi River. U.S. Army Corps of Engineers. Upper Mississippi River - Illinois Waterway System Navigation Study ENV Report 59. https://www.mvr.usace.army.mil/Missions/Navigation/NESP/Related-Documents-Info/FileId/292281/

Lines 557-563
Comment: Seasonal abundance of fish has been the focus of other studies on the UMR.

Recommendation: Add discussion regarding Johnson et al (2005) observations to bolster conclusions.  “High fish densities in the lock during the spring, summer, and fall (mid-April through November) and low densities during the winter reflect the abundance of fish in the main channel during these periods (Johnson et al 2005).”  This information seems to corroborate the study findings and should be cited.

Johnson, Brian L., Thomas M. Keevin, Eric A. Laux, and Thixton B. Miller, Donald J. Degan, David J. Schaeffer.  2005.  Seasonal Fish Densities in the Lock Chamber at Lock and Dam 25. Upper Mississippi River. U.S. Army Corps of Engineers. Upper Mississippi River - Illinois Waterway System Navigation Study ENV Report 57 https://www.mvr.usace.army.mil/Missions/Navigation/NESP/Related-Documents-Info/FileId/292279/

Lines 557 – 563
Comment:  Seasonal differences in residency time have been observed previously.  Add citation and/or discussion regarding Keevin et al (2005) findings for comparison.

Recommendation: Add discussion regarding Keevin et al (2005) observations to bolster conclusions.
 “High August mortality reflects the large numbers of fish in the lock; high December mortality possibly reflects the inability of the fish in the lock to avoid entrainment because of reduced swimming capabilities.”

“…mortality was related to water temperature, depth of water in the lock, horsepower of the tow, number of barges, and time of year.”

Keevin, Thomas M., Brian L. Johnson, Eric A. Laux, Thixton B. Miller, Kevin P. Slattery, David J. Schaeffer.  2005.  Adult fish mortality during lockage of commercial navigation traffic at Lock and Dam 25, Upper Mississippi River.  U.S. Army Corps of Engineers. Upper Mississippi River - Illinois Waterway System Navigation Study ENV Report 58.
Additional comments: 3.	 Clarifications
Line 82.
Comment: The statement “Navigation dams are known to impede fish passage in lotic systems”.  Though true, this statement is restrictive.  All dams, not just navigation dams, impede fish passage to some degree.
Recommendation:  Remove the word “Navigation”.

Lines 165 - 166
Comment: The sentence “Lock and Dam 15 has nine non-overflow roller gates and two 33-m overflow roller gates, a main lock, and a functioning auxiliary lock.” is confusing and incomplete.
Recommendation: Re-word the sentence and add a discussion of the hydropower dams. Lock and Dam 15 has 11 roller gates and two locks, a main and an auxiliary lock. It also has hydropower dams located in a secondary channel on the east side of Arsenal Island. These dams are located approximately 1.5 miles upstream of Lock and Dam 15 on either side of Sylvan Island in Sylvan Slough.

Lines 504-507
Comment: The sentence “These locks and dams have been identified as potential targets for deterrent technologies (Upper Mississippi River Asian Carp Partnership 2018) and implementation of effective deterrents at these dams could limit additional bigheaded carp range expansion in the UMR.” should be reduced.
Recommendation:  Simplify the sentence to something like; They have been identified as potential locations for fish deterrent technologies to limit bigheaded carp range expansion in the UMR (Upper Mississippi River Asian Carp Partnership 2018).

4.	Typographical comments
Lines 644-645.
Comment: UMRACT has recently changes its name to UMRICT.
Recommendation:  Change sentence to update the name change to Upper Mississippi River Invasive Carp Team, formerly known as the Upper Mississippi River Asian Carp Team.

Add volume and page numbers to line 713 and 905:
711 Camacho CA, Sullivan CJ, Weber MJ, Pierce CL. 2020. Invasive carp reproduction
712 phenology in tributaries of the Upper Mississippi River. North American Journal of
713 Fisheries Management DOI: 10.1002/nafm.10499.

903 Vallazza JM, Mosel KJ, Reineke DM, Runstrom AL, Larson JH, Knights BC. 2021. Timing and
904 hydrological conditions associated with bigheaded carp movement past navigation dams
905 on the upper Mississippi River. Biological Invasions DOI:
906 https://doi.org/10.1007/s10530-021-02583-8.

Remove parenthesis around 2018
887 Upper Mississippi River Asian Carp Partnership (2018). Potential use of deterrents to manage
888 Asian carp in the upper Mississippi River basin. http://www.micrarivers.org/wp-
889 content/uploads/2019/08/Potential-Use-of-Deterrents_Final.pdf


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: VARIATION IN THE REPRODUCTIVE QUALITY OF HONEY BEE MALES AFFECTS THEIR AGE OF FLIGHT ATTEMPT
Review round: 1
Reviewer: 1

Output:
Basic reporting: In general, the manuscript was easy to follow. It meets most of the requirements of the journal, but I had a few issues to be addressed:

I had some issues with a few complex sentences that were difficult to follow:
L85-87
L99-101
L101- Do you mean that you would expect a negative correlation between drone number and average drone size across colonies?

The Introduction was long and in some places contained unnecessary and distracting detail - the necessity of L59-66 was unclear, L66-79 seems like it could be more concise. I found myself wondering specifically about the previous study showing a reproductive advantage to large males - did they describe natural variation in size? Over what timeframe and colony sampling? Was there an idea of the range of size? There weren't many details on this study, but instead a lot of speculation about sources of variation in drone size that seem unlikely to be widespread in a natural context (queens laying in worker cells, eg). That seemed unnecessary. Similarly, parasitic feeding jumped into my mind at the outset of a source of body mass variation, and that played out here even though it received very light treatment throughout the manuscript. Perhaps you can streamline the introduction and paint a clearer picture about what is known about mass/size variation. Some of these other details could be moved to the discussion, where I would also like to see more about energetic trade-offs associated with adult body size variation and flight, larger animals have a higher mass-specific metabolic rate typically. Finally, I would like more discussion of investment in larval brood production as a function of nutritional stress. Under nutritional stress, are fewer drones produced? There is evidence that food limitation impacts worker bee physiology later in life (though maybe not body size). You could briefly mention these things and perhaps return in more depth later in the Discussion.

L75 - I don't fault the authors for this, as honey bee research has a deep history of problematic anthropomorphism, but I think the language "sloppy nursemaids" has a dual impact of sounding anthropomorphic and misogynist. Again, please don't take offense, I mean no judgement - we have a long way to go in improving language in honey bee research and behavior more broadly.
Experimental design: My biggest issue with your design and results approach is something that I think should be easy to fix - it has to do with how you report and assess "size". You report two measures of sclerotized body parts - head and thorax width. You report body mass, which encompasses both the absolute size of the animal and it's body condition (weight of soft parts, water, etc). Your result, that body mass is a predictor of emergence day while head/thorax width aren't, etc, suggest that it is in fact individual condition, not size per se, that is the best predictor of behavioral differences (and perhaps reproductive morphology differences). However, you can't definitively say that from how you report the data. I suggest calculating a true measure of condition, and re-analyzing the results following that measure (perhaps adding this to your current results if you'd like to). A condition measure is typically a measure of mass corrected for absolute body size. You would need first to determine the most accurate measure of absolute body size. There is usually a field standard, like intertegular distance, or leg length. Head width, which you already have, seems acceptable. One concern to resolve first, however, is why your measures of sclerotized body parts change over adult age, and moreover, change more the older the animal gets. I can't figure out why this would be. I recognize that animals on day 1 of adult emergence are not fully sclerotized, so their bodies could continue to grow. However, I know of no explanation for why body size would continue to change over the ~8-10 day timeframe of this study. How do you explain this? If you can resolve this problem, assessing body condition seems like the most relevant thing to do for this study. It is also a useful framework to think about trade-offs and reproductive strategies - the lighter males, at least in this study, were from colonies with more mites, which feed on developing bees (drones preferentially, as you know). The implications of variation in condition are different from those of variation in absolute body size.

Here you examine drone size variation over a very short time in the season. Throughout (including in the background) I don't see much context for the known variation in size and how that plays out over the season. I would guess there could be substantial variation with season due to many known factors (mite loads, nutritional stress) and unknown factors. Please provide more information about the role of season, and justify this short focal period.

Cell volume seems like it is a more critical determinant of size than just cell width. Is there any reason you focused on width?

Throughout the manuscript, your treatment of "foster colony" is very confusing. I think you had two colonies, one to which you introduced "AM" emerging drones, and one to which you introduced "PM" emerging drones. However, these colonies were sampled at the same time of day for drone collections. I strongly suggest changing how you label these foster colonies throughout. At present, you call them AM and PM, which is confusing. You could say "Foster Colony 1 with PM-emerging drones" or something. It is just hard to follow.

L234-236 - I think it is problematic to include the dead drones in the analysis of "flight date". These could have a variety of origins (including early death by natural causes) and it is extremely hard to interpret.

Minor things:
L154-155 - the way this is written, it sounds like you are saying you kept frames in the incubator for the duration of pre-adult development. Please clarify.
L167-168 - please explain why you did the mark recapturing with specific tags here. You do it in the Results, but it would help the reader to have it mentioned here too.
Did you account for weather variation from the time the drones were introduced, or was that an issue? That can have a big impact on even whether individuals emerge from the hive at all.
L225 - you may consider calling this "emergence" time and not "flight" time throughout because it is much more accurate as to what you actually measured. I expected some sort of measure of flight activity, duration, etc, based on the title and abstract.
L244 - you refer to the "reference colony", but it is unclear what this is and why you chose this colony. If this just means a statistical report of relative impacts of each source colony, please clarify. Any reason not to report post-hoc tests for, e.g., pairwise comparisons?
Validity of the findings: You should consider size and lifespan trade-offs in your discussion, especially since smaller males have a poorer condition. Moreover, there was very light discussion of Varroa mite feeding impacts and how those affect the interpretation of your results. Mites didn't co-evolve with apis mellifera, so that's interesting that they impact size and thus reproductive success. It is also worth considering how mite feeding may impact drone lifespan and flight capabilities. There are some studies in this area that aren't covered.

I'm not clear on your presentation of the results for mass and the relationship to mite count (L268). What is "234.54-1.06(mites)"? If this is an effect size estimate, can you please specify more clearly? (same comment applies to cell size)

L359 - I found this to be an overstatement. Even if queens mate 20x each, there is still a massive sex ratio bias towards males, and it is hard to argue this would not be a selective force, especially since colony reproductive potential is so different comparing the male and female contributions. The other components of this sentence confused me because they aren't related to drone competition. I think you are saying there are many forces that could influence drone size and constrain or weaken selection. That may be true, but don't overstate the possibility that all of these things sort of magically balance out!
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: VARIATION IN THE REPRODUCTIVE QUALITY OF HONEY BEE MALES AFFECTS THEIR AGE OF FLIGHT ATTEMPT
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript by Metz et al is an interesting topic and the results add to the literature on this topic. The English is professional throughout. The manuscript could however be improved with addition of some relevant literature as outlined in the specific comments. The figures are of good quality however figure 4 could be improved. An additional figure covering sperm count and viability could improve the story.
Experimental design: The research question of this paper was: why do drones vary widely in their size and reproductive quality?
I feel the authors were not able to answer this question. Additionally the title: Variation in the size of honey bee males leads to different life history characteristics consistent with distinct mating strategy, is not covered in the results. While the authors set out to identify mating strategies, non such were discovered or described. Non of the results presented show evidence of differences in mating strategies. A different title may be more suitable.

I have some concerns regarding sample numbers and the large variation of drones sampled between different colonies. This needs to be at least addressed in the manuscript.

I have further concerns about the criteria of what is a drone attempting mating flights. A large number of drones was dead on day 0-1 and, in my opinion, should have been excluded from the results.

Some of the methods especially for correlation analysis could have been described in more detail. individual comments in the submission are highlighted.
Validity of the findings: Some of the data on sperm viability and count is not provided. Some statistics has not been described or reported. Conclusion is not substantiated by the results or literature.
Additional comments: General comments and questions to the authors.
The manuscript was an interesting read and I did enjoy the review, however I feel the paper could be improved. I do have some significant concerns, I would like the authors to address or respond to, these are outlined in the Pdf and above.
Additionally:
The number of tagged bees in Table 1 and sheet 1 in the supplement is different. Please consider explaining of correcting.
Generally cross-referencing the figures in the results would be helpful.


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: VARIATION IN THE REPRODUCTIVE QUALITY OF HONEY BEE MALES AFFECTS THEIR AGE OF FLIGHT ATTEMPT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The Introduction is much improved and overall very clear and compelling. I had a few comments that I hope will be helpful for additional clarity. All other basic reporting requirements are met.

L47 – 48: add worker before population. Resource availability sort of, but season definitely, are environmental factors and not colony level factors. Perhaps reword the sentence to reflect that.

L 59 – it seems more accurate to say something like “their parental origin and how they are reared” (queen versus worker source of eggs isn’t really a rearing condition exactly)

L62 “laying” should be “lay”

L65 “and” should be “that”

L67 – “may be expressing” should be “may express”

L82 – 85 – this sentence was confusing to me and I’m not sure of the meaning. Perhaps reword?

L85 – 88 – I thought the way you explained this in the response to reviewer comments was much clearer than the wording here, which is confusing. For example, I think you are saying that drone fitness is impacted by selection on queen and worker traits through pleiotropy. Would this lead to “stochasticity” in drone development, or simply weaken the effects of selection on drones (weak selection in turn leads to greater variation in phenotype)? You go on to refer to the “queen-worker dichotomy”, which makes it seem like you are saying that something about the partitioning of maternal behaviors across these two castes has pleiotropic impacts on drones (an explanation that seems different and perhaps more complicated than the idea that selection on either caste impacts drones through pleiotropy). I think it would be good if you could come up with a way to word this sentence(s) a little more explicitly. One solution may be to put the sentence in L88-91 just before this.

L88 I don’t think you need the “-“ between “familial” and “social”

L92-93 – this topic sentence seems to confuse levels of analysis. Gene by environment interactions and genetic architecture problems like you described in the preceding paragraph are mechanisms that underlie adaptive responses, so they also have adaptive consequences. I think what you are arguing here is that variation in drone size could reflect a colony level reproductive strategy and that is that there may be balancing selection on drones as opposed to weakened directional selection (balancing selection could also maintain variation). Seems like the topic sentence could easily be clarified.

L104 – I appreciate the examples, but I think you could go straight into drone reproductive behavior and the possible avenues of alternative strategies (as you do about midway through the paragraph).
Experimental design: The authors addressed my previous concerns.
Validity of the findings: The authors addressed my previous concerns.
Additional comments: Very interesting study, and nicely written!


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: BIOLOGICAL AGE FOR CHRONIC KIDNEY DISEASE PATIENTS USING INDEX MODEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript addresses potentially important and useful issues in the biological age estimation.
Thus, from my opinion the manuscript can be a good contribution to the field, the style and the length are appropriate for full development of the ideas, and I therefore recommend its acceptance for publication after a minor revision.
Experimental design: Line 27: What do you mean by “intensity”? Please clarify the idea.

Lines 95-104: Correlation analysis is restricted to Pearson's correlation coefficient for quantitative variables. However, taking into account the use of categorical variables (such as gender or CKD stage), were the results obtained with other coefficients (such as Spearman's or Kendall's) analysed?

Lines 156-159: Please, explain formula (10), namely which standard deviation is used and whether BA_x and CA are from the same individual.

Line 192: Please, identify the applied statistical test.
Validity of the findings: Taking into account the formula used to compute BA estimates, does the BA index allow to estimate the biological age or only the distance between the chronological age and the biological age? That is, to assess how greater is the biological age than the chronological age?
Additional comments: Some indices are missing from formula (1)

Line 143: “I_i” should be replaced by “Index_i”

Line 166-167: Please, define \bar{d} and s.

Line 168: Please delete "Add your materials and methods here."

Lines 342-344: Improve reference 26


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: BIOLOGICAL AGE FOR CHRONIC KIDNEY DISEASE PATIENTS USING INDEX MODEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is somewhat clear, but some important details are missing or unclear. I suggest the authors provide sufficient background for a large audience.
Experimental design: In this study, the authors develop the BA using the indexing method. The results of this study show that patients with CKD between stage 2 to stage 5 experience gain in BA between 3 to 9 years.

1.	I recommend that authors use three-line tables for all tables (no vertical lines, only top, bottom, and column lines).
2.	Please indicate how the method used in this article differs from MLR and PCA. Does this method avoid the deficiencies of MLR, PCA, and KDM?
3.	Please add figure legend, a simple explanation of each picture makes it easier for readers to understand.
4.	I suggest authors use different types of statistical approaches to acquire the BA prediction models, then comparisons of biological ages calculated by the KDM, PCA, and MLR approaches to corresponding chronological ages.
Validity of the findings: The paper contributes some new ideas. But the validity of the findings in this study is somewhat limited by the small sample sizes.
Additional comments: No additional comments