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: LACUNES AND TYPE 2 DIABETES MELLITUS HAVE A JOINT EFFECT ON COGNITIVE IMPAIRMENT: A RETROSPECTIVE STUDY
Review round: 3
Reviewer: 1

Output:
Basic reporting: Review:
I truly appreciate the authors’ responses and the changes they conducted. I believe it improved the manuscript substantially! There are still a few minor issues:
1.	The authors claimed that the final enrollment number was 227, and the manuscripts were statistically analyzed based on 227 subjects. However, in figure 1 and lines 106-108, “After excluding subjects who had missing or unsatisfactory MRI data for analysis (n = 50), missing basic clinical information (n = 17), or missing cognitive assessments (n = 194), 227 subjects were included in this study (Figure 1)”. How to conclude that 227 subjects were included according to this statement and figure 1? Please calculate it again.
2.	In line 181, “…and then the appropriate t-test was used”. “t” in “t-test” should be shown in italics.
3.	Table 4 showed general characteristics of non-diabetic patients in this cohort. However, Table 4 was not mentioned in the main text after revision.
Experimental design: none
Validity of the findings: none
Additional comments: none


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

Input:
Title: LACUNES AND TYPE 2 DIABETES MELLITUS HAVE A JOINT EFFECT ON COGNITIVE IMPAIRMENT: A RETROSPECTIVE STUDY
Review round: 3
Reviewer: 2

Output:
Basic reporting: Please modify the serial numbers of other tables after deleting Table 4
Experimental design: none
Validity of the findings: none
Additional comments: none


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Dear authors,
I appreciated the reading of your manuscript.
The treated topic appears interesting; however, the novelty in your findings is not clear with respect to the previous literature. Furthermore, the introduction and discussion section mainly focus on the role of physical activity on the menstrual cycle; however, the regression analysis shows diet as the only predictor of the menstrual cycle.
I have some major concerns, which I will explain further in the following comments, and I hope they will be useful in improving your manuscript.

TITLE
In my opinion, you investigated the differences between runners and non-runners rather than the impact of recreational running. Furthermore, only diet seems to be a predictor of menstrual cycles.

ABSTRACT
1.	You did not investigate dietary patterns, but only the type of diet.
2.	What is an “A proprietary survey?”
3.	The menstrual cycle predictors appear different in the manuscript.

INTRODUCTION
1.	The FAT abbreviation is not introduced.
2.	Even though it is interesting, clear, and well written, the introduction is too long and explains details not investigated in the manuscript. The reader may lose the focus of the study and the real aim. I suggest the authors to shorten this part and introduce only the topic actually investigated in the research.
Experimental design: MATERIAL AND METHODS
1.	PARTICIPANTS:
a.	I suggest the authors be more specific about the amount and intensity of the runner group’s training. This could also be a factor to consider in the analysis as a covariate or a confounding factor. This is one of my major concerns, because women could train at different levels, intensities, frequencies, etc.
b.	I also suggest the authors be more specific in describing the control group: was it completely inactive, did it not regularly practice some kind of sports or did it train occasionally?
c.	Line 162 is a repetition of line 159.
2.	QUESTIONNAIRE:
a.	VAS should be explained.
b.	In addition to the brief explanation of the questions, you should give some information about the possible answers.
c.	Also in this case, I suggest using the type of diet or special diet and not dietary patterns, because you did not investigate dietary habits or the consumed food.
3.	STATISTICAL ANALYSIS:
a.	It is not clear if you used the SPSS version 21 or 27.
b.	Lines 196-198 are superfluous.
c.	Description of the normality assessment is not reported.
d.	Why did you opt for an ordinal regression rather than a linear one?
e.	You did not specify which variable for menstrual disorders has been used as a dependent variable and which variables have been considered predictors.
Validity of the findings: RESULTS:
1.	Table 2: why it is reported gender differences in the caption?
2.	I suggest commenting on all the significant or primary outcomes, because sometimes, in the discussion section, you explain some results without speaking about them in the result section.
3.	The chi-square test is usually indicated with the symbol χ2.
4.	In the regression analysis, you should report the results of the model (if the model is significant or not) and then the p of the single predictors inserted in the model.
5.	Line 249: in table 5, only a special diet, not time without menstruation, seems to be a significant predictor.
6.	Lines 250-255: this investigation is not reported in the study aim and the methods section.

DISCUSSION:
1.	As previously said for the introduction section, I suggest the authors be more focused on their results and not excessively digress on topics not investigated through the questionnaires. Also in this case, the reader could lose the main findings and the novelty of your investigations.
2.	In line with the previous comment, I suggest the authors reflect on the special diet as the only predictor of the menstrual cycle, while running has no effects. The authors should discuss this outcome.
3.	Line 384: Female Athlete Triad is not abbreviated.
Additional comments: No additional comments


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: I think that the present paper has an important topic and a really well-intentioned endeavor that could help the literature to fill some of the gaps existing on this matter. I'm also positively impressed by the numerosity of the sample size.

However,  a deep revision of the English language, which, I think. it is one of the main issues of this paper, is needed. Indeed, even if I overall got the meaning of most of the sentences, the syntax, structure, typos, and choice of vocabulary are extremely poor.

The material and methods section is also poorly described (i.e questionnaire section) not allowing the reader to understand much about the questionnaire and the participants. Additionally, why did you decide to use an invalidated questionnaire, you need to address this.

The discussion section should avoid reporting the results again. Also, I found that the flow of speech is not very linear and sometimes I had a hard time following.

Generally, all sections need to streamline and be more concise and clear.
Experimental design: look above
Validity of the findings: As above mention, the paper has a really good potential to be an excellent paper, however, the actual form is too primitive to be published.
Additional comments: I found quite irritating to start the reading of the paper and find a BIG TYPO on the first sentence such as FEMALE ATHLETE THRIAD!


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 1
Reviewer: 3

Output:
Basic reporting: General comments:
I reviewed the article “An assessment of the impact that recreational running has on potential disturbances in the regularity of women’s menstrual cycles – a symptom of the Female Athlete Triad”. The article aims to evaluate the impact that recreational running has on menstrual cycle disorders comparing a group of runners with a group who did not regularly practice sports (control).
The present study indicated that the runners had shorter and less regular monthly cycles and shorter and more often painless menstrual bleeding comparing with control group. In addition, it seems that the frequency of menstrual disorders in runners was increased by following a special diet even if few details were reported about diet.
The manuscript is interesting and the topic has the potential to be interesting and useful from a scientific point of view but need to be improved in all its sections in order to understand better the research.

INTRODUCTION
I think that the introduction is too long especially referring to the endocrine and energy availability components. I suggest to summarize this part reporting only what is really important for your research.
Experimental design: MATERIALS AND METHODS
Physical activity:
Referring to the aim of the study, details of the recreational running are missing. We obtained a lot of general information from the questionnaire but we don’t know the features of each session of running. How many sessions are on short or long distance? What is the real length of one session? It is conducted outdoor or on the track?

Questionnaire:
Line 181: is better to specify that this survey is for runners and I think is not correct to report “intensity of training” because you don’t properly assess it.
Validity of the findings: RESULTS
I suggest to add another table to report all the information about water and special diets (lines:232-238) even if the authors have conducted two different analysis.
Line 210 you have to report also the BMI as a result.
Line 213: add table 1 and 2 in the brackets.
Line 223: more or less pain? In the table 2 you reported painless and here pain. Please verify the data.
Line 226: specify that the incidence of menstrual pain throughout the bleeding period was less frequent in the runners even if there are not difference between the groups.
Line 231: add table 2 in the brackets.
Lines 239-242: I suggest to report and discuss all the significant correlations and not only the age. Specifically, line 239 specify that the age is the main one factor influencing the monthly cycle in both groups but not the only one. Lines 240-241, the results about the amenorrhea and menstruation pain are not referring to both groups but only for runners.
Line 249: I suggest to remove the period “extended the length of time without menstruation”.

TABLES
Table 1: add the number of participants of each group and specify under the table the statistical analysis and the p value (<0.05). Correct the unit of measure of BMI.
Table 2: complete the caption. Specify under the table the statistical analysis and the p value (<0.05). What do you mean with gender differences? Specify what is “a or b”, “x or y” under the table.
Table 3: add the number of participants.
Table 4: add the number of participants and correct the unit of measure of BMI.
Table 5: correct the unit of measure of BMI.

DISCUSSION and CONCLUSIONS
As I suggest in the introduction, I think is better to focus the discussion only on your findings without digress on other aspects.
Lines 289-290: control your data because previously you say that cycles of 25-31 days are common in both groups.
Line 295: correct into “from 25 to 31 days”.
Line 333: add in the runner group after correlation analysis.
Lines 356-357: pay attention to say that the use of special diet was conducive to the disappearance of the menstrual cycle. Actually you don’t assess properly the nutritional habits of the runners or which diet has more or less effect, so I think is too strong as a conclusion.
Line 404: change “more often” with “not”. Also in the conclusions, I suggest to not be too assertive on the role of the diet.
Additional comments: No additional comments


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors could consider to add a figure or substitute one of the tables with a figure.
it is not mandatory.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I suggest the authors insert only the useful abbreviation; for example, BMD or IOC, used only once, could not be abbreviated.
Furthermore, it would be best to be more consistent with the introduced abbreviations. Indeed, you used Group R and Group C only in the "Participants" section, while in the remaining text, you used runners and non-runners.


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Congratulation for the big commitment and the exquisite work done in addressing all the comments, I do like the new version a lot.
Experimental design: okay
Validity of the findings: okay
Additional comments: No additional comments


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 2
Reviewer: 3

Output:
Basic reporting: I have no additional comments to do.
Experimental design: I have no additional comments to do.
Validity of the findings: RESULTS
I think is a good idea to put the data referring water and special diets in table 1 even if the authors are not convinced.
In addition, pay attention because at the beginning of the results you report body weight with numerical data of BMI and not both the results (body weight and BMI).
Additional comments: I received the revised manuscript for review. I'm glad to see that the authors took reviewers' advice into consideration. The current manuscript is better than the one before it.


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 3
Reviewer: 1

Output:
Basic reporting: ok
Experimental design: ok
Validity of the findings: ok
Additional comments: ok


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

Input:
Title: THE FEMALE ATHLETE TRIAD—THE IMPACT OF RUNNING AND TYPE OF DIET ON THE REGULARITY OF THE MENSTRUAL CYCLE ASSESSED FOR RECREATIONAL RUNNERS
Review round: 3
Reviewer: 2

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No 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: DEVELOPMENT AND VALIDATION OF A MULTI-PARAMETER NOMOGRAM FOR PREDICTING PROSTATE CANCER: A RETROSPECTIVE ANALYSIS FROM HANDAN CENTRAL HOSPITAL IN CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript proposes a new nomogram to predict prostate cancer from clinical parameters. The manuscript is well written and quite clear. Also, the methodology used to build the nomogram is adequate.

I have some minor concerns:

1)	Author compares their nomogram only with local models, the reason is that international models did not fit well in their population, but no analysis of this was done. To be sure of that, it is necessary to validate PCPT in their population.
2)	Author declares that the calibration of the model is good, I agree with them, but the basic properties of the calibration, the intercept and the slope must be comment.
3)	The comparison of the nomogram with local models was performed using the entire cohort, but it must be done using separately the development and the validation cohort, otherwise, the model developed in the population is being favored.
4)	Authors used the youden index to choose a cutoff point to classify prostate cancer. This choice treats false positives and false negatives equally. It is necessary to go further, it is important to know for different cut-off points how many patients with prostate cancer are not going to be correctly classified and how many biospies could be saved. You can use the clinical utility curve for this purpose. (see Borque-Fernando Á, Esteban LM, Celma A, Roche S, Planas J, Regis L, de Torres I, Semidey ME, Trilla E, Morote J. How to implement magnetic resonance imaging before prostate biopsy in clinical practice: nomograms for saving biopsies. World J Urol. 2020 Jun;38(6):1481-1491. doi: 10.1007/s00345-019-02946-w. )
Experimental design: The study design was adequate, it would only be necessary to clarify whether or not some type of matching was carried out using a propensity matching score or similar for the choice of the validation cohort.
Validity of the findings: The use of the nomogram is limited to a local population and only uses clinical variables, but the authors declare that this is its purpose and it may have a clear utility in their healthcare setting.
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: DEVELOPMENT AND VALIDATION OF A MULTI-PARAMETER NOMOGRAM FOR PREDICTING PROSTATE CANCER: A RETROSPECTIVE ANALYSIS FROM HANDAN CENTRAL HOSPITAL IN CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: General comments
Dr Libin Nan et al reported that development and validation of a multi-parameter nomogram for predicting prostate cancer: A retrospective analysis.
Their nomogram is expected to provide benefits in making decisions to prostate biopsy in Hebei, China.

Major points

1.	Please consider adding the part “ nomogram from one institution in Hebei China” to the title because the title is very simple to misunderstanding.
2.	In the Material and Methods, about accuracy of prostate biopsy, please describe number of doctors who performed prostate biopsy and whether they were urologists.
3.	In the Material and Methods, please explain and describe prostate biopsy protocol (transrectal or transperineal, number of target biopsy etc).
4.	In the Material and Methods, please explain and describe accuracy of the pathological diagnosis more detail. (Number of pathologists, central pathologist or not, uropathologist or not etc).
5.	In the Material and Methods, generally BMI is not considered as parameter of multiparametric risk model for detecting prostate cancer. Therefore, please consider the test of multicollinearity of risk factors including BMI.
6.	In the Resutls, please explain and describe why there was a statistically significant difference only in %fPSA even though it was randomly divided into a developing group and a validation group.
7.	In the Discussion, in lines 202-204, Cut-off value P=0.31 is the ‘statistically’ best detection probability for the accuracy of the model. However, it cannot be ‘clinically’ interpreted as recommending a biopsy when the P value is 0.31 or higher and monitoring below that.

Minor points
1.	In the Material and Methods, In line 71 and 72, median is considered to be a more appropriate statistical expression than mean.
2.	The notation of statistical P value should be expressed in italics.


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: DEVELOPMENT AND VALIDATION OF A MULTI-PARAMETER NOMOGRAM FOR PREDICTING PROSTATE CANCER: A RETROSPECTIVE ANALYSIS FROM HANDAN CENTRAL HOSPITAL IN CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: I appreciate the effort that authors made to improve the manuscript, all my concerns have been amended
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: DEVELOPMENT AND VALIDATION OF A MULTI-PARAMETER NOMOGRAM FOR PREDICTING PROSTATE CANCER: A RETROSPECTIVE ANALYSIS FROM HANDAN CENTRAL HOSPITAL IN CHINA
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have adequately responded to the comments of the reviewers and improved the manuscript. Therefore, I recommend publication. Lastly please check and correct the following contents.

1. In the Material and Methods please list the citations for the biopsy protocol in order to use this nomogram in China and maintain the similar rate of cancer detection. Because it is necessary to adopt the same biopsy method.
2. In the Material and Methods, in line107 please correct the spelling　of “Smirov”. The correct name is “Smirnov”.
3. In line135, 138 and Table1-3, The notation of statistical P value should be expressed in italics.
4. Please number the figures according to the order of the annotations on the figures in the manuscript. Aren’t the numbers of figures of CUC 3 and 4 instead of 8 and 9?


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: PHYSIOLOGICAL STRESS AND HIGHER REPRODUCTIVE SUCCESS IN BUMBLEBEES ARE BOTH ASSOCIATED WITH INTENSIVE AGRICULTURE
Review round: 1
Reviewer: 1

Output:
Basic reporting: It’s always nice to see an ecological stoichiometry project in a terrestrial system, and this is one of a small handful of such papers focused on bee species that I am aware of.

There is a high degree of copy, pasting, and extremely minor editing/rearranging from previous articles from the group. In the methods this introduces errors to the manuscript (e.g., “flies”, L161, https://doi.org/10.1093/cz/zoaa070). In the introduction it results in the first and second half of the introduction having extremely different voices and depth of scientific background. In one paragraph describing the study design only the year and the temperature of freezing are changed, even though they are supposed to be two different studies performed in two different years. While I could understand for repeating methods across studies, the authors do not even refer to the other manuscripts in such a way as would be typical for similar methods or study designs (e.g., “methods were performed following Citation (Year), and thus described briefly here”). This is particularly stark in lines 62-78, 143-150, and part of the “Bumblebee Body C and N Content” heading. The degree to which it is done in this manuscript is simply unacceptable, and borders on plagiarism especially when considering that most of the copy and paste is from an article which was published this very week (https://doi.org/10.3389/fphys.2021.696689). Given that I found at least two sources here makes me concerned that there may be more I missed as well.
Experimental design: From reading the statistical methods, it seems that you used simple individual variable comparisons for each model as opposed to full models including all explanatory variables? This choice needs to be very carefully explained, as the models then do not reflect the study design and may be dramatically and incorrectly influencing the results. Further, on Gamma and Poisson distributions: Poisson makes sense to me for the latter models as those variables are clearly count data. I would add if you checked models for overdispersion and how you addressed that if present. Why was Gamma selected for the first set of models? Furthermore, there are extreme inconsistencies between the description of your analyses in the methods and the reported statistics. See line comments.

For the models that feed into your first paragraph in the Body Mass result (and Figure 1) it is unclear how you combined/split castes. It seems as though you lumped workers, gynes, and queens all together in some analyses but not others? Interchangeably using sex and caste for bumble bees is inappropriate; caste should be the focus given how biologically different workers vs. queens can be. Using sex for some analyses and caste for others makes understanding the results rather confusing. I’d suggest having the castes together as a factor with four levels (workers, gynes, queens, males). This would allow you to streamline your models, stats reporting, and effectively combine figures 1-3. Something like a Tukey HSD would then very easily determine differences among castes and between habitats.
Validity of the findings: The discussion and conclusion sections are insightful and well-written, though I anticipate significant changes will follow methodological clarity. I do think that the findings from this study could be interesting, and the discussion’s focus on the trade-offs between bee health and reproductive output is a high-point and could be an important finding in the bee world. Other authors have discussed these ideas, but the dataset presented here is uniquely poised to directly answer questions of stoichiometric trade-offs in a social insect. However, given the methodological issues as well as the presence of blatant copy/paste, it is not acceptable for publication.
Additional comments: Line Comments:
L50: Opening sentence focusing on freshwater resources and fisheries does not fit the narrative of the study well. The rest of the opening paragraph is sound. Alternatively, more comparison against previous non-pollinator ES work later in the discussion could help make this a more broadly-accessible and widely-read manuscript.
L73-78: Thank you for making the ES -> nutrition/health link. Most papers in this vein do not explicitly make this critical connection.
L80-89: Filipiak has a few other recent papers that would fit well here:
https://doi.org/10.3390/insects9030085 and https://doi.org/10.1038/s41598-020-79647-7
L80-100: While not framed specifically within ES, there’s a few other recent publications with %N in bee species which are likely to be relevant for you, and would help fill out this later section of the introduction: https://doi.org/10.1007/s00442-019-04577-9 and https://doi.org/10.1007/s13592-016-0480-4

L103-108: Your argument that the apple orchards would provide better resources than the oilseed rape fields would vary widely based on location and the type of apple orchard. For example, in other countries apple orchards are often highly intensive agricultural environments, much like how almond orchards can be extremely stressful for honey bee colonies. Thus, I think your hypotheses are dependent on the context of the study areas described in your methods, and I would consider moving your hypotheses into your methods section or providing some of that system-specific context alongside your hypotheses in the introduction. Furthermore, as an organism’s ecological stoichiometry is derived from their food source, it would help to discuss the stoichiometry and/or nutrition of the nectar and pollen sources present within and near orchards vs. rapeseed farms (you do mention this in the discussion; and it may be worth moving that here). While yes, complex habitats support a more diverse assemblage of bees, that relationship does not necessarily relate to ES.

L131: repeated ‘partially’

L143-150: This paragraph is copied nearly verbatim from another publication in your group, with only the year and temperature changing. I’d go through and ensure that these details are accurate: https://doi.org/10.3389/fphys.2021.696689

L161: Whole ‘flies’?

L172: bumblebee sex or caste? Later sentence says caste.

L180: “enclosed”

L182: Why was hive ID included for previous models but not these? I think it would be essential to include in these models, as reproductive output could depend strongly on the colony.

L195-299: The methods suggest that all models reported in this section used the same general framework and distributions. However, the test statistics reported vary widely suggesting a range of different analyses; I see chi-square, z statistics, F statistics, t statistics, as well as the omission of any such reporting (some with only a p value). Please review your models and revise.

L303-314: Likewise, I am not aware of a Poisson GLMM which returns a chi square statistic as the test statistic for individual parameters. Are these comparisons of different models? Please make clear in the methods.

L379: I don’t think the results clearly make this determination. As you discuss in the previous paragraph, bees were likely more stressed in rapeseed fields yet they had greater reproductive output. The arguments on trade-offs are much more closely aligned with your results.

L381-385: The phenology of the main flora of your habitats is an extremely important point, though I would expect the surrounding habitat would be more influential in the orchards due to the smaller patch size.

L389: Relating results to tropical flora does not seem particularly relevant to the system. The hypothesis presented in this paragraph is interesting though.

Minor thing:
“Submitted” articles are cited a few times. Make sure this lines up with PeerJ guidelines:
“Cite articles accepted for publication as 'in press'. Include in reference section and upload as a Supplemental file.
Cite work unpublished, in preparation or under review as 'unpublished data'. Supply the author's first initial and surname, and the year of the data collection, in the text citation and do not include the citation in the reference section. Example: (A Castillo, 2000, unpublished data).”


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

Input:
Title: PHYSIOLOGICAL STRESS AND HIGHER REPRODUCTIVE SUCCESS IN BUMBLEBEES ARE BOTH ASSOCIATED WITH INTENSIVE AGRICULTURE
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is an interesting study that makes a novel contribution toward understanding how environmental stressors might affect body elemental ratios of carbon (C) and nitrogen (N).  The study is nicely motivated by appropriate conceptual theory.  The data appear to be appropriately analyzed.  The MS is clearly written.
Experimental design: I have several comments and questions that need to be addressed in order to better judge the reliability of the data and the conclusions drawn from the study.

1) The MS claims that oilseed rape fields are more stressful environments than are apple orchards.  But given that the study uses commercially reared (as opposed to wild bees) bees both kinds of habitat could be considered stressful to individuals that come from commercial sources.  So, is there independent information or evidence from this or previous studies showing that commercial bees placed in these different kinds of environment are indeed differently stressed?  For example, do this or other studies show elevated metabolism in one habitat relative to another, or altered immune-response in one habitat vs another?  Offering more insight about the potential level of stress that these bees experience in the different environments would strengthen the case that body C:N differences are indeed due to stress.

 2) Do C and N contents of pollen and nectar differ between oilseed rape and apples? Could this explain the differences in bee C:N between the habitats instead of stress experienced in the different fields?  The fact that bee colonies in oilseed rape fields have higher fitness indicators might suggested that the stress hypothesis is not tenable.

3) How long were the colonies left out in the respective fields?  The reason I ask is that if the colonies were out longer than the 10-day flowering period of apple orchards, where would the apple orchard bees get food for the remaining time in the field.  If they get food elsewhere or from other plants, how much would this influence the measure of C:N for bee colonies placed in apple orchards and fitness measures?

4) Line 154: It is unclear what “post-freezing” means.  Were the collected colonies placed in a freezer to kill all of the adult and developing bees before analysis?

5) The Results section lists a bunch of statistical analyses on various attributes of bees and bee colonies.  But the Methods section provides extremely vague explanations of how the various attributes were measured.

How was bee body mass measured?

How were individuals processed in the lab to extract measures of C and N contents.  Was pollen removed from the bodies of worker bees before analyses?  Were gut contents removed from individuals before they were processed to measure body C:N?

What is the purpose of measuring the number of worker and queen cocoons and pollen and wax cells? What do more or less quantity of each of these various measures mean? Explaining this would help the general readership of PeerJ that is not familiar with bee colony life-history.  Also explain better how these variables were measured and why?  For example, are these fitness indicators?

Why were different caste members and sexes analyzed separately for C:N.  Do each potentially face different stress?  Please explain more the rationale for doing this.

The Discussion tends to be a bit more speculative about the potential that oilseed rape fields are more stressful than apple orchards, as opposed to the Introduction section which sets the contrast up as an a priori test of hypothesis about the consequences of different stress on C:N stoichiometry.   Given my comments above, more information needs to be provided to offer a definitive a priori test of the stress hypothesis (i.e., rule out competing explanations).  This would be ideal: I like tests of hypotheses.  In the absence of being able to provide such information, the MS cannot provide a definitive test.  Consequently it should be rewritten to be more speculative, that is, to explore the potential for the different habitats to be differentially stressful based on measure of C:N stoichiometry.  This would be less ideal but perhaps the only option.
Validity of the findings: The findings could make a novel contribution to the field if my concerns of alternative explanations are addressed.
Additional comments: Minor comments:

First sentence of the Introduction is superfluous.  Delete it as it has nothing to do with this study.

Line 102: “habitat quality” is vague.  What defines habitat quality for bees?

Line 109: The MS refers to Krams et al. (submitted) here and in many places throughout the text.  It would help the reader if the MS explained a bit more what that Krams study was about and what it found about C:N. that is germane to this study.

Line 405:  This explanation is rather convoluted and hard to follow and contradictory to the main thesis of the MS.  Especially so, because the C:N data presumably indicate that oilseed rape fields might be more stressful for bees, yet here the reasoning makes the opposite case that it may be a better environment than apple orchards for bees to be active.  So which is it? A stressful or not so stressful environment?


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: PHYSIOLOGICAL STRESS AND HIGHER REPRODUCTIVE SUCCESS IN BUMBLEBEES ARE BOTH ASSOCIATED WITH INTENSIVE AGRICULTURE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Overall the manuscript is very close. I have a few remaining comments below.
The expanded detail and context in the introduction is also very helpful and improves the framing of the study.
Experimental design: Thank you for expanding the methodological detail, it is now much more clear which analyses were used.
Validity of the findings: I believe I must have been unclear in my response regarding the grouping of sex v. caste for analyses. My point is that as social insects, queens, gynes, and workers are going to have different body composition (as you demonstrate in Figures 2 and 3) and thus grouping the three female castes together as a single “female” level to compare against drones (Figure 1) misses a great deal of nuance. Similarly, having queens and gynes analyzed separately from workers misses important differences. For example, if you consider the values plotted for the C/N ratio panels (Figures 2D and 3D), “young queens” appears to be similar to workers whereas “oviposting queens” alone may be driving the reported difference between workers and queens. Likewise, drones may be different in their C/N composition from some castes of females but not others, as has been seen in other species of social insects (e.g., https://doi.org/10.1111/j.1365-2435.2009.01604.x , https://doi.org/10.1007/s00040-012-0220-3). Body mass is another variable where it does not make biological sense to group all females together, with the massive size variation from workers to queens that is possible even within single bumble bee colonies. As presented, the findings suggested from the analyses underlying Figures 1 and 2 may only exist as a result of the decision to group multiple castes of females together in different ways for different models. Performing your analyses with a caste factor with four levels (drone, worker, gyne, queen) would tease apart potential differences among them, simplify your analyses by reducing the number of models and figures, more accurately reflect their functional and biological differences, still answer your primary questions, and allow this manuscript to have greater reach among entomologists studying other social insects. Your response to a comment by the other reviewer shares the same rationale: “Moreover, workers, queens, males, young queens and ovipositing queens have different social, functional and reproductive roles in the hive and outside the hive. “
Additional comments: L110: Missing word: “Based these…”
L351-353: This sentence on the phenology of oilseed rape v. apple trees misses the habitat context: that the former is an agricultural monoculture and the latter is a more complex, patchy habitat with many other food resources and edge habitats.
L354-356: You did not measure rate of growth, only final reproductive output.
L356-359: Phrasing this as a question is strange as you answer the question earlier in the paragraph. Would be more effective and interesting to introduce the tradeoff idea here before the next paragraph: indication of stress in oilseed rape field, yet more reproductive output in oilseed rape fields.
L384-387, 389-396: I think the approach of trying to give a simple answer of which habitat is “better” doesn’t match the results. Individual bees are more/less stressed in one vs. the other, while colonies are more/less reproductive in one vs. the other. Both habitats are “better”, from a certain point of view. Clearly the truth is somewhere in the middle. Furthermore, a limited study where purchased colonies were only on the ground for a few months should be careful to make broad conclusions. The strength of this work is your ability to assess the tradeoffs of ES stress vs. reproductive output in bees, and I would focus on that in these first few paragraphs and then the closing of the discussion. It’s a truly novel contribution and I would really work to make that the focus of the discussion.
L398: Strike ‘unlimited’
L398-413: This hypothesis is interesting but, particularly after the revision, feels extremely disconnected from the rest of the manuscript. I’d either remove it or move a brief mention of the hypothesis (1 sentence?) to the following paragraph.


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: PHYSIOLOGICAL STRESS AND HIGHER REPRODUCTIVE SUCCESS IN BUMBLEBEES ARE BOTH ASSOCIATED WITH INTENSIVE AGRICULTURE
Review round: 2
Reviewer: 2

Output:
Basic reporting: Improved based on my comments in the original review.
Experimental design: Improved based on my comments in the original review.
Validity of the findings: Improved based on my comments in the original review.
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: INFLUENCE OF LYMPHANGIO VASCULAR (V) AND PERINEURAL (N) INVASION ON SURVIVAL OF PATIENTS WITH RESECTED ESOPHAGEAL SQUAMOUS CELL CARCINOMA (ESCC): A SINGLE-CENTER RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The article is well written. It has scientific value. We can accept the article.
Experimental design: The article is well written. It has scientific value. We can accept the article.
Validity of the findings: The article is well written. It has scientific value. We can accept the article.
Additional comments: The article is well written. It has scientific value. We can accept the article.


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: INFLUENCE OF LYMPHANGIO VASCULAR (V) AND PERINEURAL (N) INVASION ON SURVIVAL OF PATIENTS WITH RESECTED ESOPHAGEAL SQUAMOUS CELL CARCINOMA (ESCC): A SINGLE-CENTER RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors analyzed and demonstrated the relation between LVI/PNI and the survival rate of ESCC. However, the writing quality for this manuscript is not acceptable for this journal.

There are many grammar mistakes including the misusage of punctuation and unclear statements. This main manuscript including the figure legends needs to be carefully edited with assistance from a scientific writer with appropriate scientific English writing skills.
Experimental design: The experimental design is well proposed with well-described statistic analysis.
Validity of the findings: There is sufficient data to support the conclusion in this manuscript.
Additional comments: Please see below for some minor comments:

1.	The authors need to present the full name of a term in the first instance and follow it immediately by the abbreviation in parenthesis. For instance, the authors need to use the full name for the median OS on line 35.
2.	The authors need to add scale bars in Figure 1.
3.	Do not use abbreviations in the title.
4.	The font size of the references doesn’t match with the rest of the manuscript, please enlarge them.
5.	The authors need to use consistent formatting in the main manuscript when citing other people’s work.


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

Input:
Title: INFLUENCE OF LYMPHANGIO VASCULAR (V) AND PERINEURAL (N) INVASION ON SURVIVAL OF PATIENTS WITH RESECTED ESOPHAGEAL SQUAMOUS CELL CARCINOMA (ESCC): A SINGLE-CENTER RETROSPECTIVE STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: Line 62: Formatting inconsistencies: some spaces between text and parenthesis, others don't have spaces (e.g Line 62)

Line 79: recommend rephrasing this sentence

Line 134: Figure1 is not showing prognosis, it should be Figure2

Line 137: If the authors can reformat the figures and put the number of patients in each subgroup and the number at each sensor time point  below the figure, that would greatly improve readability and clarity for the data and analysis

Line145: It will be a lot easier if the figures2 are labeled with panel numbers such as A, B, C, D (like Figure3 and 4), and in the main text, each result refers to a specific panel. Also in the legend, have a short description for each panel.

Line153, Line 156: grammar issue, cross out "were"

Line 157: in Figure3 legend, I recommend for authors be more precise on the subtitles for each panel instead of just putting down a P value.

Line 169: it was stated some studies reported that the LVI is not only a negative prognostic factor independent of tumor stage but there is only reference. Recommend adding additional references where appropriate.

Line 177: missing references for other studies when authors stated their results were the same as that in other studies.

Line 205: Please clarify what this sentence meant.
Experimental design: Line 159-161: This part needs more details and clarity. First of all, the title says Univariate and Multivariate Analysis of Survival, but is the P values reported here based on univariate analysis or Multivariate Analysis? Second, based on the table3 of the univariate analysis, there are other variables that also show a significant p value, such as invasion but are not reported in the main text? Does the invasion here indicate both V1N1? Please clarify. Third, what the multivariate analysis shows is not reported in the main text of the result section.

Line 165: the results need some clarity. Did the authors only look at V1N1 patients or all patients including three other subtypes?
Validity of the findings: Line 154-157: the interpretation and results do not match. The result shows only in V1N1, patients who underwent chemo had better OS. There were no direct comparisons between the histological types. I recommend authors re-write these parts to match results.

Line 214: The conclusion that post-operative adjuvant chemotherapy should 214 be done for the patients with V1N1 is too strong and not justified without proper discussion and weighing the benefit and risks involved.
Additional comments: I found many grammar mistakes throughout the manuscript and recommend English proof-editing.


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: INFLUENCE OF LYMPHANGIO VASCULAR (V) AND PERINEURAL (N) INVASION ON SURVIVAL OF PATIENTS WITH RESECTED ESOPHAGEAL SQUAMOUS CELL CARCINOMA (ESCC): A SINGLE-CENTER RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have addressed my concerns with the original manuscript.
Experimental design: The authors have addressed my concerns with the original manuscript.
Validity of the findings: The authors have addressed my concerns with the original manuscript.
Additional comments: I just have some minor comments about the formatting:

1. For reference (line 343-345), the format should be "Hamanaka R, Yokose T, Sakuma Y, Tsuboi M, Ito H, Nakayama H, Yamada K, Masuda R, Iwazaki M. Prognostic impact of vascular invasion and standardization of its evaluation in stage I non-small cell lung cancer. Diagn Pathol. 2015 Apr 2;10:17. doi: 10.1186/s13000-015-0249-5. PMID: 25884820; PMCID: PMC4413537." instead of "Rurika, Hamanaka, Tomoyuki, Yokose, Yuji, Sakuma, Masahiro, Tsuboi, Hiroyuki, and Pathology IJD. 2015. Prognostic impact of vascular invasion and standardization of its evaluation in stage I non-small cell lung cancer. Diagn Pathol 10 1-10 DOI 10.1186/s13000-015-0249-5."

2. Please add spaces between text and parenthesis, for example, line 59, 61, 63, 65, 67, 69, 71, 73, 75 in the first paragraph of Introduction 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: INFLUENCE OF LYMPHANGIO VASCULAR (V) AND PERINEURAL (N) INVASION ON SURVIVAL OF PATIENTS WITH RESECTED ESOPHAGEAL SQUAMOUS CELL CARCINOMA (ESCC): A SINGLE-CENTER RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Much improved and addressed all my previous comments and suggestions. Therefore no more comments.
Experimental design: Much improved and addressed all my previous comments and suggestions. Therefore no more comments.
Validity of the findings: Much improved and addressed all my previous comments and suggestions. Therefore no more comments.
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: RELIABILITY AND VALIDITY OF THE MULTI-POINT METHOD AND THE 2-POINT METHOD’S VARIATIONS OF ESTIMATING THE ONE-REPETITION MAXIMUM FOR DEADLIFT AND BACK SQUAT EXERCISES
Review round: 1
Reviewer: 1

Output:
Basic reporting: This study aimed to examine the concurrent validity and reliability of the multi-point method and the two-point method’s variations for estimating the one-repetition maximum (1RM) in two exercises: the deadlift and squat exercises. Participants performed six sessions of 2 incremental loading tests: multi-point method and two-point method variations, followed by 1RM tests. The authors conclude that the multiple-point method and most of the two-point method load variations examined in this research provided (from large moderate to perfect) an accurate estimate of the 1RM. Therefore, they recommend using the multi-point method and specially two-point methods variations, including higher relative loads in estimating 1RM.
The introduction of the manuscript is sufficient, and appropriate bibliographical references are included.
Figures and tables can be considered adequate and clear.
Experimental design: Numerous studies have been done lately to estimate the value of 1RM, but, to our knowledge, in this case, a set of predictors not previously used are applied.

The methodology is well described. However, the procedure for determining the reliability of the measures should include the intraclass correlation coefficient (ICC) and the coefficient of variation (CV) based on the total root mean square within-subjects of the repeated measures ANOVA analysis.

On the other hand, the mean velocity (MV), not the mean propulsive velocity (MPV), is used for the analysis of the load-velocity relationship. It is known that at low percentages, the MPV gives slightly higher values than the MV. This circumstance may result in the best fit of the percentage-velocity relationship not being a straight line but a curve. Although the authors conclude that high loads should be used as a reference to estimate 1RM, the results could be different if the MPV is used.

When the percentage values used for prediction are very different, a major problem can occur. For example, when the subject jumps from 60 to 90% of 1RM, he may not be in the best condition to obtain his maximum performance at 90% since he has not received sufficient prior stimulus to move such a high load. Likewise, it is unlikely to perform to the best of the subject's ability at 60% without prior warm-up. If any lifting is performed prior to 60% and some intermediate between the two percentages, it should be indicated in the methodology. This warm-up is necessary to arrive in the best state of potentiation to the high loads. The best example is the progressive increase of loads that weightlifters perform in training and competition. These previous lifts do not have a negative effect by fatigue but a positive effect by potentiation.

In the discussion section, we have not seen reference to the analyses with the Bland-Altman plots.
Validity of the findings: The following reasons could condition the validity of the results:
1. The CV values are very high in both exercises. It does not seem reasonable to recommend the application of a procedure that does not present sufficient reliability.
2. If a warm-up has not been used prior to the lower of the percentages and between the two percentages, likely, the performances were not the maximum that the participants could achieve.
3, As the authors themselves indicate, the value of 1RM can change frequently. Therefore, the recommendation to use any of the procedures indicated would require performing a 1RM test every day or at least very frequently since the absolute loads representing the different percentages could have changed.
4. It should be explained why, for example, the correlation obtained with 40-90% is 0.839, 60-80% is 0.062, and 60-90% is 0.872. The study suggests that the higher the loads, the more accurate the prediction. Therefore, the relationship obtained with 60-80% should be at least similar to the other two indicated
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: RELIABILITY AND VALIDITY OF THE MULTI-POINT METHOD AND THE 2-POINT METHOD’S VARIATIONS OF ESTIMATING THE ONE-REPETITION MAXIMUM FOR DEADLIFT AND BACK SQUAT EXERCISES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Reliability analysis should be improved.
Experimental design: Excellent!
Validity of the findings: They are OK, but reliability analysis should be performed again and absolute errors in 1RM prediction shouls also be provided.
Additional comments: Authors should be congratulated for a general well-written article and the examination of two basic exercises less investigated in the VBT literature. The article is interesting but I have two main considerations: (I) I believe that reliability was not properly evaluated (see comment below) and therefore all the discussion related to the reliability indices should be modified after doing the proper calculation, and (II) absolute errors should be added to the validity analysis. Some specific comments can be found below:

Line 34. Replace “consisted in” by “consisted of”.

Line 115-116. Replace to: “to identify the optimal combination of the two loads that allows predicting the 1RM with the highest accuracy”.

Method is OK but I think it would be clearer if divided in different subsections (participants, study design, procedures, and statistical analyses).

Line 186. It is not clear how the CV was calculated for reliability analyses. CV should be calculated as the standard error of measurement (typical error) / participants´mean score x 100). I believe the CV value provided in this version of the manuscript depicts the between-subject variability instead of the within-subjects variability, and this is the reason why authors found a poor reliability but in fact I think that the reliability has not been analyses yet because the current computation of the CV is not appropriate.

Table 1 and 3. authors should also provide the absolute errors.

Table 2 and 4. The huge CV also for the real RM is very strange. This confirms that reliability has not been calculated properly.

Line 262. Which minimal velocity threshold did you use?

Line 271. García-Ramos and Perez-Castilla did not recommend loads below for 1RM prediction. They have recommended loads below 40%1RM for assessing the F-V relationship which are different things. The differences between both relationships should be better explained here, and clarify that the 2 optimal loads depend on the relationship to be determined: F-V requires a point close to v0 and another close to F0, and L-V the lower point always recommended by García-Ramos and Perez-Castilla is approximately 50%1RM and the higher should be closer to 1RM (aprox. 85%1RM). The rest of the arguments I think they are quite solid!

Line 293. And this can be checked with your data, which is very interesting, when you do the proper reliability analysis!


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: RELIABILITY AND VALIDITY OF THE MULTI-POINT METHOD AND THE 2-POINT METHOD’S VARIATIONS OF ESTIMATING THE ONE-REPETITION MAXIMUM FOR DEADLIFT AND BACK SQUAT EXERCISES
Review round: 2
Reviewer: 1

Output:
Basic reporting: Dear authors, thank you for your compliments regarding the usefulness of my previous review. My main two points was the proper calculation of CV values and the calculation of the absolute errors for determining the accuracy in the estimation of the 1RM. Authors indicated that new CV values were added to tables 2 and 4, but I do not see any chanhe implemented in these tables. Results in the text are changed but in the tables are the same. Now the results presented in the table do not match the results presented in the Tables. In addition, authors indicated that they added the absolute errors in the estimation of the 1RM for the different models, but I was not able to see these results in the revised version of the manuscript.

It is very important to me to implement these changes clearly in the revised version of the manuscript.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No comment


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

Input:
Title: RELIABILITY AND VALIDITY OF THE MULTI-POINT METHOD AND THE 2-POINT METHOD’S VARIATIONS OF ESTIMATING THE ONE-REPETITION MAXIMUM FOR DEADLIFT AND BACK SQUAT EXERCISES
Review round: 3
Reviewer: 1

Output:
Basic reporting: Ok
Experimental design: Ok
Validity of the findings: Ok
Additional comments: Ok


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: RELATIONSHIP BETWEEN THE DISTRIBUTION OF VEGETATION AND THE ENVIRONMENT IN THE COASTAL EMBRYO DUNES OF JALISCO, MÉXICO
Review round: 1
Reviewer: 1

Output:
Basic reporting: Edaphic characterization of the coastal dunes of Jalisco, México, and its relationship with vegetation distribution (#63863)
Introduction
The introduction section has several paragraphs that some of them should be merged (e.g., lns: 101-102 is a sentence!!!). Also, all citations used in the introduction is a bite old. Also, after reading the introduction, it is impossible to understand the gap and consequently the innovation of the paper.
Overall, introduction needs to improve by adding the background related to the problem statement of the study by using updated citations.
Experimental design: Materials and methods:
The sampling site (Lines 147-154) names should be added to the map of sampling sites.
Please clarify the date of sampling.
In the Laboratory section why only the sand fraction has been measured? As we know the silt or clay fractions can have strong correlation with chemical properties like OM.
Based on lines 205-212 the title of the manuscript should be changed to Environmental instead of edaphic!!!!
Statistical and spatial analysis: I think when we decided to investigate the relationship between environmental properties and vegetation types, we need to use multivariate statistical methods like CCA, DCA, or PCCA to show the real relationship. These methods combine both variables and it would be easy to determine the controlling parameters.
Validity of the findings: Results:
In the first section of results there are a set of correlation coefficient values and associated p-values, maybe it is better that bring them in a Table! And remove several paragraphs! The main point here is that all coefficients are weak correlations (less than 0.6; although the p-value is significant). When we review these results, we can see that sometimes 60% of data cannot explained by these coefficients: so, in discussion it should be explained, and the reasons must be found.
Table 1: the communalities is correct NOT Commonalities!!!
You should bring all PCs loading values as well as engine values, explained variance,…have a look on similar papers. Merge Table 1 &2.
Clustering the environmental shows that cluster 2 only has one site that you mentioned you collected 1- 2 samples in each site. So first you have to mention the number of samples in each cluster and although with 2 samples the statistical analysis should be meaningless.
Table 4: Group!!!!
Fig. 5&8: what is the meaning of the letters?
Additional comments: Discussion and conclusion: several paragraphs that I could not find the rational relation.
Overall, I think this manuscript needs a lot of works to be improved and suitable for publication.
Good luck


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: RELATIONSHIP BETWEEN THE DISTRIBUTION OF VEGETATION AND THE ENVIRONMENT IN THE COASTAL EMBRYO DUNES OF JALISCO, MÉXICO
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: See attached 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: HIGHLY PATHOGENIC AVIAN INFLUENZA VIRUS OF THE A/H5N8 SUBTYPE, CLADE 2.3.4.4B, CAUSED OUTBREAKS IN KAZAKHSTAN IN 2020
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript by Amirgazin et al entitled, “Large outbreaks in wild birds and poultry in Kazakhstan during 2020 caused by the highly pathogenic avian influenza virus of the H5N8 subtype, clade 2.3.4.4b” is a study on H5N8 isolated from dead birds in Kazakhstan. The investigators performed whole genome sequencing and their analysis showed that all isolates were from the clade 2.4.4.4b and were related to other isolates from nearby geographical locations. It is the first report of clade 2.3.4.4b viruses in Kazakhstan and expands our awareness and knowledge of this lineage of avian influenza viruses. To increase readability, it would be great to have a copy-editing service, especially for parts of the Discussion. Specific comments are below:

Line 20: If I understand correctly, it should be “hit hard” instead of “hit hardly”. The phrase “hit hardly” refers to few occurrences while “hit hard” refers to many occurrences.

Line 45: The high pathogenicity avian influenza (HPAI) designation is based upon disease outcomes in gallinaceous bird species (e.g. chickens, turkeys, quail). In most cases (with exceptions), HPAI viruses only cause mild clinical disease in wetland species. It would be good to specify “flocks” as “gallinaceous flocks”.

Line 47: “sever” to “severe”

Lines 47-48: “~50% chances of death” to “high case-fatality ratio”. The reports of fatality rate are usually based upon known IAV spillover event to humans. We do not fully know how much spillover occurs as IAV is not routinely tested in the hospital/clinical setting. The use of high case-fatality ratio is thus more appropriate here.

Lines 50-52: Reassortment does occur between avian and mammalian viruses, but they are uncommon. More common reassortments are between strains from the same host. I think this sentence should also reflect the likelihood of these reassortment events.

Line 55: “ and HA (9 subtypes)” to “and NA (9 subtypes)”. Also please define HA (hemagglutinin) and NA (neuraminidase).

Line 64: “hardly hit” to “hard hit”

Line 80: Please consider using a different word in place of “epiornithia”.
Experimental design: The data and analyses presented are sound and appropriate. The methods are well-presented. Below are some specific points to address:
Line 150: “month” to “months”
Line 155: eliminate “purpose”
Line 160: Does the word “subjected” mean that the samples were also submitted, processed, analyzed in other laboratories? If so, I think a more detailed/technical word should be replace “subjected”.
Validity of the findings: 1.	Can you be more specific with the location each strain? Are there any epidemiological links with some of the isolates?

2.	The clade/cluster assignments need to be reconsidered since some are paraphyletic, i.e., it does not include all descendants of the branch. Specifically, K8 should also include isolates from K3 and K6 since all share a common ancestor. It would thus be best to assign K8, K3, and K6 as one cluster.

3.	For the discussion of mutations and associated biological outcomes, it would be best to mention that additional experimentation would be needed to determine if the same outcomes would be observed for the H5N8 Kazakhstan isolates. The genetic background plays a big role in whether a biological outcome will be achieved (also called epistasis). Indeed, it is possible to have a mutation and observe different outcomes depending on which genetic backbone is used by the virus.

Below are some specific comments:

Line 179: For influenza literature, the term “clade” has a specific definition which refers to the grouping of HA sequences according to the guidelines of the H5 evolution working group (See reference below). Therefore, it’s good practice to avoid the use of the term clade unless it refers to the HA grouping. I think it would be best to use terms such as “cluster” or “group” instead of “clades” to avoid confusion. If you perform interclade pairwise comparisons and find a >1.5% difference between isolates, then you can assign presumptive clades but eyeballing Figure 2, it appears that the Kazakhstan isolates do not satisfy these criteria.

•	World Health Organization/World Organisation for Animal Health/Food and Agriculture Organization (WHO/OIE/FAO) H5N1 Evolution Working Group. 2014. Revised and updated nomenclature for highly pathogenic avian influenza A (H5N1) viruses. Influenza and Other Respiratory Viruses 8:384-388.

Figure 2 caption: “Only bootstrap values no less than 60 are shown” to “Only bootstrap values greater than or equal to 60 are shown”

Figure 3 caption:
•	What is rationale behind using a bootstrap cutoff of 60 for the HA and 59 for the NA sequences?
•	Are the clades/clusters indicated by K1, K2, etc. according to the HA groupings? If so, please indicate in the caption explicitly.
Additional comments: Nothing further to add


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

Input:
Title: HIGHLY PATHOGENIC AVIAN INFLUENZA VIRUS OF THE A/H5N8 SUBTYPE, CLADE 2.3.4.4B, CAUSED OUTBREAKS IN KAZAKHSTAN IN 2020
Review round: 1
Reviewer: 2

Output:
Basic reporting: In their manuscript, Amirgazin et . al.  present a study  describing the detection and full genome sequencing of several A/H5N8 samples from Kazakhstan in 2020. A/H5N8, and other HPAI, are a global concern and the details from this paper are highly important for understanding global circulation and impact of these viruses.  However, the authors could greatly improve the data included in the study and further refine the paper for overall readability and novelty.
1.	Overall, the paper could use an edit for English language, grammar, syntax, and tense such as line 20, 34, 47, 48, and forward
2.	Line 55: Technically there atre 18 HA subtypes and 11 NA subtypes including influenza virus family members found in bat species
3.	Influenza subtypes should be reported according to generally accepted nomenclature including type as A/HxNy or A(HxNy) wherever used in the manuscript
Experimental design: 4.	In terms of the phylogenetic analysis,  a Maximum Likelihood methodology may be much more robust and could them be coupled with a TMRCA analysis in BEAST or similar bioinformatics analysis.
5.	In addition to the HA and NA, internal gene segments of avian influenza viruses  are equally important for understanding reassortment, geographical spread, and even zoonotic risk. It is critical to add analysis of internal genes of these viruses as well as a tanglegram describing their relation to other global strains.
6.	Individual strains are not generally considered to be individual clades, please consider revising
7.	Please include a table of all mutations detected in each strain for comparison across samples described in this manuscript
8.	Please ensure to include some type of indicator or text  for subtype (ie: A/H5N8, A/H5N6, etc..) and GISAID or other database accession numbers in the phylogenetic trees
9.	Was any  further in vitro or in vivo work performed or planned to confirm pathogenicity or differences with these isolates?
Validity of the findings: No issues were determined with validity of the findings
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: HIGHLY PATHOGENIC AVIAN INFLUENZA VIRUS OF THE A/H5N8 SUBTYPE, CLADE 2.3.4.4B, CAUSED OUTBREAKS IN KAZAKHSTAN IN 2020
Review round: 1
Reviewer: 3

Output:
Basic reporting: Amirgazin et al., sequenced H5N8 HPAI sampled in Kazakhstan and conducted phylogenetic analysis for sequence comparison to the other H5 viruses from public database. Although the study is important in terms of an epidemiologic standpoint, data presentations are not constructive, and the structure of the manuscript needs to be significantly improved and organized for readability.
Experimental design: The methods described are not sufficient enough to cover what are shown in the results.
Validity of the findings: Throughout the manuscript, the author misused the term used in influenza virus research, and the naming of the clade classification of GsGD H5 lineage virus does not follow the recent WHO guidance.
Additional comments: line by line comments

Line 43: The two references used to demonstrate the natural reservoir of AIV should be replaced with other references. They are not comprehensive, and one of them is irrelevant.
Line 44: Poultry also can be infected with IAVs, but not all IAV. Generally, IAVs that can infect and productively circulate in wild birds and poultry are called Avian influenza virus (AIV). It is also defined by the tripartite international organization (WHO, FAO, and OIE). Throughout the manuscript, the authors should have used the term, Avian Influenza virus (AIV), not IAV.
Line 50: Again, IAV represents all influenza type A viruses.
Line 71-72: If the outbreaks are still ongoing and declared HAIV free, state it more clearly as of the time this manuscript has been written.
Line 75: What the recent prototype sequences mean is unheard of. If you used previously isolated sequences from the public database such as GISAID or IRD, make it more transparent.
Line 76-77: What is more important thing than that specific subtype (H5N8) has never been detected in Kazakhstan is that the A/Goose/Guangdong/96 (Gs/GD) H5 lineage has ever been introduced to Kazahstan. Has ever Gs/Gd H5 lineage virus been detected in Kazahstan before?
Line 77-78: Determination of basic amino acid in the HA cleavage site is the result of this study.
Line 84-85: What were the species of dead birds? How many dead birds were sampled? Was there any clinical complications found at the time of dead birds found?
Line 119: Does the identifier present the accession number given by Genbank or GISAID? Then, add strain name along with the number.
Line 125, 128: Using the sequences deposited during time doesn’t make sense. It should be isolated.
Line 143-145: Could you provide an approximate number of wild birds affected during the die-off event?
Line 149: What is the calculation of the number of affected poultry? Were there any measures taken, such as culling poultry from the affected farm? In that case, does the 2 million include the culling?
Line 151: Eleven out of fourteen?
Line 157-158: Since 1976 out of 2212 carcasses tested positive, what was the criteria that the ten viruses were picked up for whole genome sequencing?
Line 157: because we don’t know if the viruses sampled were viable by only doing the RT-PCR and the viability was not confirmed by egg inoculation, the term ‘isolate’ should’ve been used. This term is found throughout the manuscript.
Line 159-164: The other viruses sequenced elsewhere is not part of this study. Other sequences used for phylogenetic analysis should be in the materials and methods.
Line 167-168: Looking at the phylogenetic grouping, the H5 subclade classification used in this study referred to some previous papers published when early clade 2.3.4.4 viruses emerged (clade 2.3.4.4 group A – clade 2.3.4.4 group D). However, this classification was updated in detail as the viruses diverged. Please use the latest classification (clade 2.3.4.4a – clade 2.3.4.4h) as the factsheet (https://apps.who.int/iris/handle/10665/336259)
Line 174: How many nucleotide differences were found in NA?
Line 175: Why are there 16 NA sequences? Only ten viruses were sequenced in this study.
Line 179-190: For the Gs/Gd H5 lineage, the clade classification means the sub-lineages of H5 HA genes. The K1 – K8 naming should be cluster. In addition, looking at the phylogenetic HA and NA trees some clusters does not supported by a bootstrap value (generally bootstrap value >70 is acceptable for clustering).


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: HIGHLY PATHOGENIC AVIAN INFLUENZA VIRUS OF THE A/H5N8 SUBTYPE, CLADE 2.3.4.4B, CAUSED OUTBREAKS IN KAZAKHSTAN IN 2020
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript by Amirgazin et al entitled, “Highly pathogenic avian influenza virus of the A/H5N8 subtype, clade 2.3.4.4b, caused outbreaks in Kazakhstan in 2020” presents the detection of clade 2.3.4.4b H5N8 high pathogenicity avian influenza viruses in Kazakhstan from various avian species. Phylogenetic analysis of the sequences demonstrate that the viruses sequenced were related to viruses isolated in neighboring countries. This manuscript is a major improvement over the prior version in methods used for phylogenetics, content, and writing style.
Experimental design: The use of Bayesian phylogenetics is a much appropriate method to analyze avian influenza sequences. The description of the methods used are sufficiently detailed to be able to reproduce the results.
Validity of the findings: The assignments of putative phylogenetic groups according to the HA tree are more appropriate although it is unclear what is the epidemiological significance of these groupings. Do the samples in each HA phylogenetic grouping share a common geographic location and/or species from which the samples were taken from? Is the linkage temporal? I would suggest addressing the epidemiological significance of the B1-5 groups in the discussion 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: REMARKABLE GENOMIC DIVERSITY AMONG ESCHERICHIA ISOLATES RECOVERED FROM HEALTHY CHICKENS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The goal of this manuscript was to survey the diversity of Escherichia in a single layer flock using a combination of traditional isolation coupled with genomic approaches. The authors found that, by screening a population of ~100 isolates, there was a very diverse population of Escherichia harbored by a relatively small number of birds. These isolates appeared to be primarily non-pathogenic, as they lacked virulence genes typical of ExPEC and APEC, and they no isolates belonged to the notorious B2 phylogenetic type typical of ExPEC. Overall, this work is a nice snapshot of the diversity of the genus harbored by birds and is useful information.
Experimental design: Can the authors please describe the farm used to obtain the Lohmann layers? Some more information would be relevant because the focus is on Escherichia diversity and ecology, such as flock size, barn design, ventilation, and were the pullets for this flock sourced from multiple flocks or a single flock?

Some clarification on Table 1 would be helpful. Why did the number of primary isolates collected differ by date collected? Age of bird in the table would be helpful as well. Is number of unique isolates based on the GTG5 PCR? It was unclear if additional isolates were screened and filtered into those characterized in Table 1, or if this table represents all isolates screened?

For the custom APEC database, some of the genes listed are really not APEC virulence factors and are instead ExPEC virulence factors derived from the work of James Johnson. I would suggest that this list of genes be refined according to more recent APEC literature, which focuses on some of the genes on the list but emphasizes the presence of the pathogenicity-associated island on ColV-like plasmids, which is established the strongest link between virulence potential in birds, and is of highest prevalence in clinical isolates and most differentiating between clinical versus commensal isolates.

Figure 1 could be used to further clarify where the GTG5 screen falls into the workflow?

Figure 2, definition of the colors should be clearly defined in the legend (what are they, specifically?). Also, no scale is provided for the tree.

Figure 4, title should be plasmid replicon contents, not plasmid contents as the approach used only surveyed replicons.

One of the most enlightening aspects of this manuscript was the E. coli ChromeAgar clearly grows other Escherichia. This should be further discussed in the manuscript or brought to attention of the reader as it has substantial implications for researchers.

While the primary focus of the manuscript was on the substantial diversity of E. coli even in a setting that is fairly uniform and controlled, the authors do discuss the absence of the B2 phylotype and its implications re: commensal   E. coli populations in the layer gut. Since this was brought up, some limitations of the study need to be addressed in the study, including N=1 flock, small numbers of isolates and timepoints, small number of birds, etc.

Is there any way to better present the diversity of Escherichia harbored by a single bird at a single timepoint? This would be useful.
Validity of the findings: The findings of the authors that extensive diversity exists were supported by the genomic data. I question if the study design was robust enough to make any conclusions about the ecological makeup of Escherichia in the layer gut, that could be generalized more broadly. I would suggest that the authors take caution in some of the discussion points focusing on APEC in the gut, and how an N=1 flock translates to layers wordwide.
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: REMARKABLE GENOMIC DIVERSITY AMONG ESCHERICHIA ISOLATES RECOVERED FROM HEALTHY CHICKENS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper of Thomson et al. analyses the genome sequences of 100 isolates of Escherichia coli from fecal samples of 24 healthy adult chicken. The work is globally well done. The main finding is that, in this small sample size of hosts, a huge genomic diversity is found with representative of the major phylogenetic lineages of the genus Escherichia except E. albertii, Shigella and E. coli phylogroups B2 and G. This is indeed remarkable, as stated in the title.

Here are some remarks to improve the manuscript.

The authors should look to the serotype (O:H) and fimH diversity of the strains. They should also search for intestinal virulence genes.

The authors should discuss the fact that APEC strains are mostly found in G (ST117) and B2 (ST95) phylogroups, which are absent from their data set.

Lines 284-85, the authors are right, but there are some tools that can help and should be tested (see doi: 10.1099/mgen.0.000398).

Lines 366-67, not true. See Clermont et al. Environ Microbiol, 2011, clades are frequently isolated in birds.

Lines 28-29, please rephrase in a more comprehensive way without abbreviation.

Line 114, LB is  for Lysogeny Broth (not Luria Bertani).
Experimental design: OK
Validity of the findings: OK
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: REMARKABLE GENOMIC DIVERSITY AMONG ESCHERICHIA ISOLATES RECOVERED FROM HEALTHY CHICKENS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have addressed all concerns of reviewers #1 and #2 nicely. I have no further concerns. This is a nice and useful paper.
Experimental design: The authors have addressed all concerns of reviewers #1 and #2 nicely.
Validity of the findings: The authors have addressed all concerns of reviewers #1 and #2 nicely.
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: REMARKABLE GENOMIC DIVERSITY AMONG ESCHERICHIA ISOLATES RECOVERED FROM HEALTHY CHICKENS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The paper has been improved and can be published.
Experimental design: OK
Validity of the findings: OK
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: VALIDITY OF USING PERCEIVED EXERTION TO ASSESS MUSCLE FATIGUE DURING RESISTANCE EXERCISES
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comments. See PDF
Experimental design: No comments. See PDF
Validity of the findings: No comments. See PDF
Additional comments: No comments. See 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: VALIDITY OF USING PERCEIVED EXERTION TO ASSESS MUSCLE FATIGUE DURING RESISTANCE EXERCISES
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: 1.  I believe the paper could benefit from including more data on volume.  Volume was determined as  number of repetitions performed at 65% 1RM, and number of repetitions was determined as 30, 60 and 90% of maximal repetitions from an initial maximal repetition test.  Is that correct?  It might be interesting to report the mean, SD and range for repetitions at each level for each exercise, and for the maximal repetition test.
2.  Do you think performing the maximal repetition test immediately prior to experimental trials might have affected exertion?  Could the maximal repetition test have been performed in the orientation session?  Why did you choose to do anchoring procedures with the maximal repetition test rather than with the 1RM test?
3.  Lines 187-190 indicate that some participants had difficulty maintaining the pace.  The authors explanation of what occurred in this situation could use some clarification.  Weren't participants already pausing between repetitions?  Were participants only struggling to maintain pace during the concentric portion?
4.  The authors use the terms "velocity loss" and "velocity changes" throughout the paper.  As velocity is included in the prediction models, it might be helpful to more clearly differentiate between the two terms, and to clarify the velocity that is used in the models vs. "velocity loss".
Validity of the findings: no comment
Additional comments: This paper contributes to the literature in this area and I appreciate the goal of facilitating determination of fatigue for practitioners.  The results could be very applicable in practice.

1. In the abstract, line 30, suggest changing the word "exhibited".  Resulted in?
2.  I found lines 72-76 a bit confusing; perhaps reword?  I understand the idea of velocity loss validity during only explosive exercise, but then it is unclear how the next statement is an example?  Are you saying that for hypertrophy, explosive exercises are less ideal and so velocity loss is not a good indicator of fatigue in that setting?  Perhaps the issue is just with the term "for example".  Maybe "However"?  Or I may be misunderstanding...
3.  Line 86 - what is meant by the "shape" of the RPE scale?  I was curious on your choice of the CR-10 scale... so perhaps this is a place to explain that choice?  There are also a variety of task-specific OMNI scales, although it seems those weren't used here, which is fine, but could use an explanation.
4.  Do the authors feel that RPE alone reflects fatigue?  Or only in conjunction with velocity?  Line 366 might suggest that RPE alone reflects fatigue - perhaps some number above 8 on the CR-10 scale?  That is the general suggestion for aerobic activities.  Could the authors comment on the use of RPE alone?
5.  In lines 379-394, I might suggest different words for "unignorable" "un-failure".  I'll admit I don't have great suggestions and ultimately it makes sense.  Perhaps reword lines 381-2 ... I think the authors are saying that the experimental design of non-explosive, single joint activity was not suitable for using velocity loss as a measure of fatigue... and therefore, exercise in that type of setting would not benefit from using velocity loss. Perhaps un-failure could be reworded as "not to failure" design.  Not sure that's any better :).


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: VALIDITY OF USING PERCEIVED EXERTION TO ASSESS MUSCLE FATIGUE DURING RESISTANCE EXERCISES
Review round: 2
Reviewer: 1

Output:
Basic reporting: All of my comments and concerns have been addressed. The newest edition of the paper is sufficient for publication.
Experimental design: All of my comments and concerns have been addressed. The newest edition of the paper is sufficient for publication.
Validity of the findings: All of my comments and concerns have been addressed. The newest edition of the paper is sufficient for publication.
Additional comments: All of my comments and concerns have been addressed. The newest edition of the paper is sufficient for publication.


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

Input:
Title: VALIDITY OF USING PERCEIVED EXERTION TO ASSESS MUSCLE FATIGUE DURING RESISTANCE EXERCISES
Review round: 2
Reviewer: 2

Output:
Basic reporting: No Comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The changes made by the authors in response to reviewers strengthens the paper and addresses major concerns.


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: CHARACTERIZATION OF AMPHOTERIC BENTONITE-LOADED MAGNETIC BIOCHAR AND ITS ADSORPTION PROPERTIES FOR CU2+ AND TETRACYCLINE
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this manuscript, 8 biochar adsorbents were prepared and used to adsorb Cu and tetracycline. The topic is interesting. However, the mechanism analysis is not enough and the structure is not good. Therefore, "Major revision" is recommended. Specific comments are provided as follows:
1.	The abstract is too long, it needs to be further simplified and highlight innovation. People care about the new mechanisms or extraordinary performance more.
2.	The introduction was too short. Why the authors choose Cu and tetracycline as adsorbates? the authors were suggested to talk more about the Cu and tetracycline, including source, hazard, treatment method comparison, adsorption amount, etc.
3.	Line 56, two spaces.
4.	Line 86, how the amphoteric modified bentonites of “B (0%BS-B), 50%BS-B and 100%BS-B” were prepared? What’s 100%BS-B, no bentonite was used?
5.	Section 1.1.1, please combine the sub-paragraphs into one paragraph.
6.	How the pH value of solid sample was measured?
7.	Space between value and unit was missing, such as line 101, 102. Double check the full text.
8.	line 110-112, mL or ml? r/min or r min-1? Notice the color of text and commas, such as line 112-113, and line 141-145.
9.	line 131-132, poor format.
10.	line 166, bentonite (B), no need to abbreviate again.
11.	The meaning of annotation in Figure 1 needs to be explained.
12.	The resolution of SEM image is too low, so it needs to be updated. In addition, the ruler and other information in the image are not clear.
13.	What is adsorption rate? How to calculate?
14.	The author suggested that Fe3O4 particles block the pores of biochar, well then, why the surface area of MC was larger than C?
15.	The governing mechanisms were unclear, which should be emphasized. XPS analysis needs to be added to support the adsorption mechanism.
16.	Most of the cited references are out-of-date. It is suggested to cite the up-to-date literatures to highlight the significance and novelty of this research.
17.	The English needs to be improved. The manuscript needs to be well organized. For example, combine the paragraphs with similar contents, and add the topic sentence at the beginning of each paragraph.
18.	The conclusion needs to be more concise.


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: CHARACTERIZATION OF AMPHOTERIC BENTONITE-LOADED MAGNETIC BIOCHAR AND ITS ADSORPTION PROPERTIES FOR CU2+ AND TETRACYCLINE
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this paper, amphoteric bentonite was loaded onto magnetic biochar in a novel direction and the adsorption effect of Cu2+ and tetracycline in water was studied. Authors used few characterization techniques to support their material properties, but fails to explain the data critically. Moreover, authors just focused on some preliminary studies, and they did not go in depth. Anyway, it could be re-considered for publication in this journal after the following revisions to improve its quality before publication.

1.	The abstract was written like conclusion, please rewrite and state some key findings of your study.
2.	The introduction is not well structured and less of clarity, especially paragraph 2.
3.	The author should give a very clear introduction of BS-12, which is a major modifier in the experiment, like its amphoteric property.
4.	There are some scientific, grammatical and typo errors in the manuscript that need to be re-checked and corrected more carefully. E.g., Line 47, “many repair materials studied more” should be “many repair materials were studied”; Line 56, two spaces between clay and using. Some sentences are too long. Others were highlighted in PDF.
5.	All the figures are not clear, and the resolution are too low. I highly recommend that the authors should use different line types for the four lines in figure 3, not only use different color to distinguish them. Plus, what’s the meaning of Bb, Cc, Aa… above the columns in Figure 1?
Experimental design: 6.     The author should explain the experimental methods more detailed, like the heating rate and holding time for the preparation of biochar by pyrolysis. The meaning of the samples’ name should be clarified, such as 0%BS-B, 50%BS-B and 100%BS-B.
Validity of the findings: 7.	Line 156-158, a lower CEC value means better ion exchange ability? Magnetic modification and the addition of BS-B promoted reduced the CEC value while promoted the ion exchange?
8.	The conclusion in section 3.1 are not that firm. If the BET of the samples is the key character to determine the adsorption effect of TC, then MC should show the best adsorption capacity because it has the highest surface area. There is no doubt that BET is very important for adsorption, but there must be some other properties also play a vital role, like CEC, although the correlation is poor. The authors also discussed the effect of CEC on adsorption in section 2.6. So I doubt the significance of the correlation analysis.
9.	The MC100%BS-B has a lower BET and CEC, while has the highest qm for Cu2+, why?
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: CHARACTERIZATION OF AMPHOTERIC BENTONITE-LOADED MAGNETIC BIOCHAR AND ITS ADSORPTION PROPERTIES FOR CU2+ AND TETRACYCLINE
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors have made several biochar/bentonite composites, and also investigated the adsorption of Cu and tetracycline on these materials. In general, the English language of the whole manuscript is very unclear and ambiguous. There are many language defects throughout. The next most important issue is the inaccurate material naming. For example, the name of carbon-based amphoteric bentonite (CBS-B), should be a modified bentonite, not a biochar. So the abbreviation CBS-B seems to be ambiguous. Another problem is incomplete experimental data. For example, the authors calculated the adsorption thermodynamic parameters, but they did not conduct the thermodynamic adsorption experiments.
Experimental design: .Line 88, “and then magnetized by coprecipitation method loading Fe3O4 to prepare MC....” what is the detail method?
.Line 114, please provide the method reference.
Line 122, Is 12 hours enough? what is the adsorption equilibrium time?
Validity of the findings: Figure 1, What do the letters on the figures mean?
.Line 251-254, The data in Figure 7 and table 3 are inconsistent. The adsorption rate of 98.28% seems inaccurate, please check them carefully.
Additional comments: .The English language should be improved to ensure that an international audience can clearly understand your text. Some examples where the language could be improved include lines 21-22, 28-30, lines 96-102, etc.
.line 61-62, what is the scientific basis of this deduction?
.Your introduction needs more detail. The impact and novelty are not assessed.