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: COMPARING ENZYME ACTIVITY MODIFIER EQUATIONS THROUGH THE DEVELOPMENT OF GLOBAL DATA FITTING TEMPLATES IN EXCEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: Although the main points made in this paper are correct, the knowledge the author displays of the literature is completely inadequate, as one might guess from the fact that fewer than half of the references are to work done in the 21st century (not counting the author's own publications), and most of those that ARE from the 21st century address points of detail rather than the main thesis. Córtes et al. (2001, Biochem. J. 357, 263-268) provided a more detailed account of the relationship between inhibition constants and IC50, and referred to an earlier study by Cheng and Prusoff (1973), that considered the same question. However, even 2001 is only just in the 21st century, so the author should read Baici's recent book (2015, Kinetics of Enzyme-Modifier Interactions, Springer) as well as reading the relevant papers that he cites.

Sebaugh's paper (2011) is addressed to pharmaceutical scientists, and it is absurd to say that it "distill[s] inhibitory theory down to its most basic form". It contains nothing that a biochemist would call inhibitory theory.

The paper by Tipton et al. (2014) is OK as far as it goes, but it says almost nothing relevant to the present paper. It does, however, include references to work the author should read.


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

Input:
Title: COMPARING ENZYME ACTIVITY MODIFIER EQUATIONS THROUGH THE DEVELOPMENT OF GLOBAL DATA FITTING TEMPLATES IN EXCEL
Review round: 1
Reviewer: 3

Output:
Basic reporting: This manuscript presents a package for analyzing enzyme kinetic data by means of global fitting within an Excel template.  The author argues that the common practice of characterizing kinetic data by visual examination of graphical representations often leads to oversimplified interpretations. The routines presented in the current work enable the simultaneous analysis of raw data according to standard models of competitive, non-competitive, uncompetitive and mixed inhibition. A second template permits the analysis of the partial versions of these mechanisms. Both templates include an equation (the “Modifier” equation ) incorporating all of the other mechanisms.  Vm, Km and Ki values are derived for each mechanism as well as the RSS measure of goodness of fit. The template thus offers a rapid and objective evaluation of different kinetic models and is potentially of considerable value to the field. The Figures in the manuscript mostly consist of clips from the Excel files included in the Supplementary material and are generally helpful summaries of the voluminous information contained in the spreadsheets.
The author argues that the conventional forms of enzyme kinetic equation are too complex and obscure the mode of action of inhibitors. The equation employed for the fitting involves a rearrangement of the more familiar format  to demonstrate separate effects of inhibitor or activator on apparent Km or Vm values. To this reviewer, the rearrangement is no simpler than the more traditional forms of the equation. This does not, however, invalidate the result and an alternative view may well be helpful to some.
The design of the work is acceptable. It employs data sets from other workers and subjects these data to the global fitting procedure to compare their fit to the various models employed. At one point in the manuscript, the utility of the process for determining competitive inhibition models is queried.  The author might consider testing the validity of the process by using  simulated data with zero errors, to determine if the fitting method does actually work for  this mode of inhibition and if so, how well.
The presentation of the work is has some other problems.  There is potential confusion for the reader in the use of Km, Ki and Vm which should be regarded as constants defining the enzyme. The author frequently  refers to the variation of Vm, Km or Ki under varied inhibitor or substrate concentrations– where what is meant is the apparent or measured Vm, Km or Ki. The distinction should be made clear throughout – perhaps by using italicized terms for the experimental values. In places the wording suggests that the Km or Ki represents the affinity of substrate or inhibitor for the enzyme, whereas they are inversely related to the affinity. This should be clarified.  Sometimes arguments are presented in prose that is difficult to understand and I recommend a careful reexamination of the text to remove such issues. Instances are given below.
In the spread sheets supporting the paper the results for different types of mechanism are compactly presented in the first sheet, with detailed information about each mechanism in the following sheets. In the first sheet, numerical results are presented, with values for Vms, and various K parameters. These Tables are presented in abbreviated form as Figures in the text.  The goodness of fit for each mechanism is illustrated via v vs s and 1/v vs 1/s plots. (I note that in the sheet1 of the S2 file, double reciprocal plots appear to be flat. A scale calculation appears to need adjustment.) Choice of the best fit mechanism(s) is made easily on the basis of RSS values. In some cases the fit to different models is very close. The inclusion of standard errors on the regression parameters would help to distinguish between alternatives in such cases since it is possible that an improved overall fit might result from inclusion of an additional but poorly defined parameter.
This reviewer suggests,  perhaps for future adoption, that in the templates for partial inhibition, inclusion of Dixon plots on Sheet 1 , may illustrate the partial nature of the inhibition better than the double reciprocal plots currently shown.
In some cases (e.g. in S9) the values shown in the summary table at the top of sheet are not reflected in the tables for individual mechanisms. Thus in S9 table for the modifier equation, values for Vmax 2 and Ki appear to have been interchanged  and  Km2 has acquired the value of Vmax.
In other instances, similar confusions may be found. In S13, the modifier equation shown in the bottom box contains the term Kx, rather than Ki. A value for Ki is given in the overall summary, but this value becomes that of Vm in the summary box at the bottom of the sheet.
It is not clear whether the author wishes the global fitting routines to be generally accessible. Pasting new data into the spreadsheets results in no progress beyond the initial estimates. If it is intended that the routines become generally available, some instruction as to the use of the global fitting routine would be valuable.

Minor Issues:
Title: “ Modifier equations” rather than ” modifiers equations”
Line 27: replace “pseudo steady-state” with “Rapid equilibrium”.
Line 30: change sentence after “changes in the…. “ and substitute “measured values of Vmax and Km”
Line 35: change ‘’would be’’ to ‘’is’’.
Line 45: insert ‘’possible’’ after ‘’total” , change “turnover’’ to ‘’conversion’’
Line 77: substitute ‘’absent” for “conspicuously missing”.
Line 85: insert after “plot ” “experimentally determined values of ”
line 90: change “affinity” to “access”
line 92: insert “apparent” before “substrate” and before ”Km”
Line 94: insert “apparent value of” before “Vmax”
Line 98: change “of” to “or”
Line 102 : IC50s
Lines 113-5: Meaning of this sentence unclear.
Lines 116-119: Sentence is incomplete.
For all equations: make sure equation number is aligned with the equation itself.
Line 136 – 139: Meaning is obscure. One assumes that there is a one-to-one association of the inhibitor with the enzyme.
Line 142: processes
LinE 144:  “only affect the Km” may read better as “partially increase the measured Km’’
Line 147 – enzyme’s
Line 149: allows
Line 164: Fig 2f. does not illustrate hyperbolic behavior.
Line 167: suggest insertion of “ïn fact’’ after  ‘’Km are”
Line 168-169: suggest insert brackets around “as it could………………Kallikrein”
Line 197: Why is a modification of the direct linear plot used? Are there advantages over the original v vs s form  that can be described.
Line 224: ‘’much more even”
Lines 276-279: meaning of this sentence unclear –“ particularly strict distribution of lines”.
Line 289: “these, not “theses”
Line 347: “in that it”
Line 348: Equation 7 contains only one inhibition constant.  Is 7 the equation that is meant?
Line 362: Intended meaning of this sentence unclear – “access?”
Experimental design: Experimental design:
The work reported here involves the analysis of data obtained from other workers and thus has the merit of objectivity.  It is suggested, though, that the use of simulated data with known (or zero) variability would enable a testing of the limitations of the approach.
Validity of the findings: The presentation of this work is a little challenging, as discussed above, but there is no doubt that the analytical approach proposed may simplify and standardize the data analysis of experiments in which the mode of action of inhibitors or activators  of enzymes are the focus of the work.
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: COMPARING ENZYME ACTIVITY MODIFIER EQUATIONS THROUGH THE DEVELOPMENT OF GLOBAL DATA FITTING TEMPLATES IN EXCEL
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comments
Experimental design: no comments
Validity of the findings: no comments
Additional comments: The methodology was not described as it was suggested. The mathematical methods were not detailed. The methods section remains the same.

Please, explicitily state the objective of this work by describing: "the objective of this work was to..."


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: COMPARING ENZYME ACTIVITY MODIFIER EQUATIONS THROUGH THE DEVELOPMENT OF GLOBAL DATA FITTING TEMPLATES IN EXCEL
Review round: 2
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: In my original confidential comment to the editor I said "the author has a lamentably poor knowledge of the literature. The work itself is in general sound, but the views expressed are nowhere close to being original. Numerous previous authors have made the same or similar points". All this remains true, and now I say it directly to the author, as he doesn't seem to have picked up the implications from the comments directed to him.

The revision has been very half-hearted, and the author seems to have been more concerned to justify his own position than to address the reviewers' comments in a serious way. Here I only discuss the responses to my own report, but things could also be said about the other two.

1. "Most new research is a rehash of former work". Yes indeed, and it's particularly true of this paper. That's why there needed to be a proper discussion of previous work.

2. "The reviewer’s inability to provide a reviewed article later than 2001". Has it not occurred to the author that it's HIS job to make a literature study? Reviewers can point at useful directions to take, but he shouldn't expect them to do his work for them. It was not a matter of inability but of disinclination. I can think of at least three books (in addition to Baici's) published in the 21st century that provide some of the review that the author is missing. However, he can look for these himself.

3. No serious reason is given for refusing to cite Cheng and Prusoff. Among the numerous papers that could have been cited, but weren't, there are several from Tipton's group, such as Lizcano et al. (1996) Biochem. Pharmacol. 52, 187-195, that are relevant.

4. "Baici’s non-peer reviewed book". This insulting remark displays an astonishing ignorance of how serious publishers operate. Does the author imagine that Springer publishes books without getting them reviewed? Vanity publishers are, of course, different, but the author would have to be even more ignorant to think that Springer is a vanity publisher. Relatively few books in biochemistry make any mention of peer review (and those that do are usually general textbooks), but that doesn't mean there wasn't any. No, Baici didn't cite Fontes et al., and maybe he should have, but we can only guess at his reason. He also did not refer to Mahler and Cordes, as I would have done if I'd been writing that book, and if I were writing the present paper. Dixon & Webb, too, while we're at it, not to mention J. Leyden Webb (a different Webb). Even Botts and Morales (1954), the first, as far as I know, to discuss the general modifier mechanism.

5. "A valid opinion for biochemists." Yes. In case the author hasn't realized it, enzyme inhibition is a biochemical topic; applications in pharmacology are derivative. To dismiss the opinions of biochemists as of no account hardly deserves a reply.


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: COMPARING ENZYME ACTIVITY MODIFIER EQUATIONS THROUGH THE DEVELOPMENT OF GLOBAL DATA FITTING TEMPLATES IN EXCEL
Review round: 2
Reviewer: 3

Output:
Basic reporting: This paper concerns the creation of an Excel template  designed to analyse kinetic data and fit them to a variety of modes of inhibition or activation affecting single-substrate enzymes .The fitting process operates on the basis of minimising the sum of squared residuals and thus permits the selection of the mechanism that gives the optimal fit. This routine will undoubtedly be found useful by many in the field and merits publication. However the presentation of the material still requires considerable revision.
The introduction is long and detailed but lacks clarity. Much is made of the failure by many workers to distinguish between inhibitory effect and the inhibition constant. It is not clear what is meant by the inhibitory effect – is this the relative inhibited velocity (vi /v) or the  IC50 or a component of the inhibition equation.?  This is not clear and is nowhere defined in the manuscript. As a result , the material presented is difficult to absorb.
The introduction argues, legitimately, that the use of IC50 values to characterise an inhibitor does not define the mechanism. It is also argued that characterising an inhibition mechanism by visual characterisation of Lineweaver-Burk plotting can led to over-simplification. The Excel template offers and objective means of identifying the most plausible mechanism and giving values for the constants defining it. It would be more useful still if calculation of standard errors on the regression parameters were included. Researchers are likely to be more interested in how well individual constants (Km, Vm, Ki) are defined than in the overall goodness of fit.
Several statements in the text refer to values of Km or Vm varying linearly or hyperbolically – but with what independent variable is not stated. It is assumed that the variable is inhibitor concentration. This should be stated frequently enough to remove any confusion. Fig 2 as it stands tends to add confusion to the discussion. All changes in the value of the apparent  Vm or Km are represented as linear changes. The figure would be more helpful if they showed, for the  different mechanisms,  the variation of apparent Km and Vm values with [X] – clearly demonstrating hyperbolic or linear changes.
Since the models  employed in the template have been discussed in detail in a previous publication by the author and others, it seems unnecessary to readdress the questions in the detail found here. This section could be clarified by a) showing a reaction scheme embodying all the proposed mechanisms and b) showing the equations generated by the model. without necessarily giving a detailed derivation of each.
In summary, I believe that while   the  fitting  template is potentially of value to workers in the field, the presentation of the work still requires substantial revision.
Experimental design: No comment
Validity of the findings: The author has presented sets of simulated data to demonstrate the merits of the templates. However, the Dixon plots for the data from the partial inhibition simulations are linear. This is not what one expects for partial inhibition (in which the enzyme inhibitor complex retains some activity) . It is also notable that in, for instance, the partial uncompetitive inhibition data set, the fit to the partial inhibition model is the worst of those tried. These data sets need to be carefully checked.
Additional comments: Matters of detail:
The Km does not measure substrate binding affinity – it is inversely related to the affinity. The higher the affinity of the substrate for the enzyme the lower the value of the Km. Similarly, Ki is referred to as a binding constant for the inhibitor whereas it is strictly a dissociation constant.

Line 122: It is implied that partial inhibition mechanisms have been recognised only relatively recently although, in fact, they have been text book material for over 50 years (e.g. Dixon and Webb, Enzymes, 1964).
Line 88. “ubiquitously”
Line 119-122: This sentence suggests that mechanisms or equations are inhibitors.
Line 124 “are believed to affect the activity” would read more accurately as “represent the activity”
185-190: the intended meaning of this passage comes through only with difficulty. In competitive inhibition, the apparent Michaelis constant is given by Km*(1 +[I]/Ki) which is clearly linearly related to the inhibitor concentration.
“In the total inhibitor equations, the Ki is equated to the effect of the inhibitor on the enzymatic activity rather than an equilibrium binding constant marking the concentration where half the enzyme population is bound by the inhibitor. ” Can the author cite instances in which such a confusion is observed?


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: EVIDENCE FOR THE CRETACEOUS SHARK CRETOXYRHINA MANTELLI FEEDING ON THE PTEROSAUR PTERANODON FROM THE NIOBRARA FORMATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript is well thought out, and written well aside from a few small typos. It was easy to read, clear and concise. Within my attached review I made comments with regards to English and grammar when needed. One suggestion I have for the authors is to state where and when the specimen is from in the abstract. It is not very clear and would help in future literature searches.

The literature references were appropriate and well researched. I recommend the authors reviewing Ehret and Harrell, 2018 (which was just published) and have added a few other suggested papers, but they are not necessary per se. The figures are done well and are consistent with the content of the manuscript.

Artist's reconstruction of the potential event, while somewhat fantastical, is wonderful. The author's fully admit that artistic license was taken.
Experimental design: This original manuscript reviews the predation of a pterosaur which is preserved in a specimen from Kansas. The specimen is mentioned on the Oceans of Kansas website, and has been cited in other research papers but it has not been formally researched and described. Due to the fragile nature of pterosaur bones, preservation is not common, in particular specimens with evidence of predation. The authors do a good job describing the specimen and discussing how it came to be. The specimen is likely a composite, however, abundant evidence and citations are given to demonstrate that the vertebrae and shark tooth are a truly associated specimen. In my attached review, I suggest that the authors further investigate the orientation of the shark tooth in the specimen. Thinking about the orientation of the tooth while it was in the shark's mouth, and the mechanics of how the event likely occurred, it lends more support to the authors' conclusions. I also suggest the authors familiarize themselves more with the common terminology of shark teeth. Portions of the description were somewhat confusing.
With the few numbers of pterosaur bones present with predation/scavenging marks, this research is an important contribution to the field.
Validity of the findings: The authors describe the specimen in full and are conservative with their hypothesis. Their conclusions are well founded and researched.
Additional comments: See other comments in marked up manuscript. Aside from a few suggestions regarding references and relating tooth orientation to feeding/scavenging event, I am pleased with this report.


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: EVIDENCE FOR THE CRETACEOUS SHARK CRETOXYRHINA MANTELLI FEEDING ON THE PTEROSAUR PTERANODON FROM THE NIOBRARA FORMATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: Clear and unambiguous, professional English used throughout.

Literature references, sufficient field background/context provided.

Self-contained with relevant results to hypotheses.

Meets standards.
Experimental design: They describe the specimen, compare it to similar specimens, etc. Interpret the results.

Meets standards.
Validity of the findings: All seems valid, Cretoxyrhina bit on Pteranodon, and why wouldn't it. The only questionable  thing is the acrobatic shark snatching the pterosaur out of the air, which the authors do not suggest was normal feeding mode of Cretoxyrhina.
Additional comments: See that attached pdf for more detailed 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: ACUTE EFFECTS OF CONTRACT-RELAX PROPRIOCEPTIVE NEUROMUSCULAR FACILITATION STRETCHING OF HIP ABDUCTORS AND ADDUCTORS ON DYNAMIC BALANCE
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an original article. The article presents significance in the content, however, the description of it needs to be improved. The aim of the present study was to investigate the effects of a single dose of contract-relax proprioceptive neuromuscular facilitation stretching of hip adductors and abductors on dynamic balance in the frontal plane.
Experimental design: Research question well defined.
Methods described with sufficient details.
Statistical analysis was simplified and unspecific to this study.
Validity of the findings: The study presents interesting results, but with a limited analysis.
Additional comments: Title:
I recommend changing “frontal plane” to “medio-lateral balance”. And avoid writing your title as a question.
Abstract:
Please, add your main purpose (aim) in the abstract.
Add a rationale for your hypothesis.
Please, follow a single order (i.e. p=000.x; and d=0.0x or vice versa).
Introduction:
This section is too long and present unnecessary and basic information about stretching and postural control. Your introduction needs to be more specific.
Now, you added the aim, but forgot the hypothesis/rationale.
Methods:
Participants: Did you define the power of this study?
You have different genders in your same sample. Could you explain how anthropometric differences (i.e. pelvic width) can affect the balance?
Please, add the # of the IRB.
Procedures: Your purpose does not require “no feedback” and “visual feedback” conditions. Why did you do both?
Did you measure the intensity of the stretching bout (i.e. Level of discomfort)?
Please, add the version/model of the Libra stabilometric platform.
“To compensate for learning effects, the order of the trials was reversed in half of the sample (crossover design). Is this a crossover design? Explain.
Statistical Analysis: Why did you use a t-test if you have 3 factors (leg [right and left]; pre- and post-CR results; and with and without feedback)?
You have to describe your index to the Cohen’s d in this section.
Please, review your statistical analysis.
Results:
 You cannot use this description of the significance: p = 0.000. Change to p < 0.001. This section is confusing, and based on effect sizes.
It is not possible to trust in your results, after your Statistical Analysis section.
Discussion:
This section is too long and present unnecessary information. Your introduction needs to be more specific, and related to your results.
Conclusion:
Please, improve your conclusion, and follow your purpose.


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: ACUTE EFFECTS OF CONTRACT-RELAX PROPRIOCEPTIVE NEUROMUSCULAR FACILITATION STRETCHING OF HIP ABDUCTORS AND ADDUCTORS ON DYNAMIC BALANCE
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is an interesting study, and the effects of stretching on dynamic balance have practical relevance to fitness and rehabilitation interventions. However, the Introduction does not provide enough background and rationale for why it is expected that CR stretching would affect dynamic balance. The authors focus primarily on what balance is, what sensory systems play a role, etc. in the Introduction and very little on the mechanisms of stretching and how/where/why that would affect the control of balance. Although a background on balance is important, the authors present some information that is irrelevant and never touched upon again in the context of the current study (e.g., why do I care that there are two motor strategies [ankle and hip] to maintain balance, and how does this relate to the current study?). More emphasis needs to be placed on stretching (particularly CR stretching), how it works, how it changes the system, and the mechanisms by which this would alter the control of balance.

The authors include a lot of background research in this manuscript in both the introduction and discussion sections. However, there are many long paragraphs in which several ideas are buried, and it is difficult to know what are the main points. For example, the first paragraph in the Discussion (lines 186-219) seems to present a large number of other studies that examine stretching and balance control yet I have no idea whether I am supposed to take away that the current study is in agreement or disagreement with these prior studies. In contrast, the next paragraph of the discussion section (Lines 220-222) is only two sentences and does not provide enough detail. The organization of the Introduction and Discussion sections need to be revisited in order to enhance the readability of the manuscripts and better present both background ideas and the results of the current study.

Raw Data – The data presented in the supplemental file is well-organized and easy to follow. However, the only data given are the calculated variables (TA, EA, ET, RT, and GI) and not the underlying raw data used to get those numbers.
Experimental design: The research question is well-defined and the methods are generally described in sufficient detail and with enough information to replicate. However, I have several concerns regarding the experimental design:

1. Learning effects – The authors have already taken into account some aspects of learning effects by reversing the order of trials (feedback vs. non-feedback trials) in half of the participants. However, what the authors have no accounted for is the learning effects that may occur by simply repeating the balance task a total of four times without having performed stretching in the middle. To account for such learning effects the authors should include a control group that does not receive stretching.

2. Statistical tests – It is good that the authors tested normality of the data before selecting a parametric statistical test and including the effect sizes is a huge plus. The actual statistical tests used (dependent samples/paired t-tests) may not be the most appropriate. The authors are comparing 9 different metrics across two different conditions. Some sort of ANOVA may be more appropriate to account for the multiple comparisons and allow better comparisons between what happens in with feedback and without feedback conditions.
Validity of the findings: Given the problems stated above with the experimental design, I am not yet convinced that the results support the conclusions. Particularly because many of the p-values (especially for without feedback condition) are barely below the alpha of 0.05. Although the conclusions will likely hold (given the high effect sized with visual feedback, for example), the authors need to re-evaluate their results and conclusions after addressing the experimental design deficiencies.

The results need to be placed in better context with prior studies; addressing the prior comment regarding long, unorganized paragraphs will help place them in context.
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: ACUTE EFFECTS OF CONTRACT-RELAX PROPRIOCEPTIVE NEUROMUSCULAR FACILITATION STRETCHING OF HIP ABDUCTORS AND ADDUCTORS ON DYNAMIC BALANCE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The major remaining critique of this manuscript is the organization of the Introduction and Discussion sections. Although the Introduction has been modified to address prior critiques (e.g., irrelevant information removed, and more discussion of mechanisms of stretching), it remains hard to follow. Instead of having one long paragraph, the authors should split the Introduction into several smaller paragraphs, each focused on a main point that helps build the rationale for the study. The Discussion section is similarly plagued by a long paragraph in which it is hard to identify the main points. Reorganizing both the Introduction and Discussion sections is necessary to enhance the readability of the manuscripts and better present both background ideas/rationale and the results of the current study.
Experimental design: The resubmitted manuscript is much improved by inclusion of a control group and adjusted statistical analyses.
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: ACUTE EFFECTS OF CONTRACT-RELAX PROPRIOCEPTIVE NEUROMUSCULAR FACILITATION STRETCHING OF HIP ABDUCTORS AND ADDUCTORS ON DYNAMIC BALANCE
Review round: 3
Reviewer: 1

Output:
Basic reporting: The Introduction and Discussion sections of the manuscript are much improved. I suggest the following minor edits:

- Line 45: Replace “Balance control have” with “Balance control has”
- Line 56: Replace “The changes in the muscle-tendon unit stiffness” with “Changes in muscle-tendon stiffness”
- Line 63: Replace “Two of the PNF techniques” with “Two PNF techniques”
- Line 83: I don’t think Versteeg et al. 2016 addresses ML balance (as it is an AP model of balance control).
Experimental design: The research question is well-defined and the methods are generally described in sufficient detail and with enough information to replicate.
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: THE INFLUENCE OF MATERNAL BODY MASS INDEX AND PHYSICAL ACTIVITY ON SELECT CARDIOVASCULAR RISK FACTORS OF PREADOLESCENT HISPANIC CHILDREN
Review round: 1
Reviewer: 1

Output:
Basic reporting: Review line 72 "being overweight'

Raw data was not shared
Experimental design: No comment
Validity of the findings: Explain the use of threshold stated in line 201 as standard for inclusion in multiple regression model i.e. reference

Assuming that the variable used in models are confounders of the association-how was this determined?

Indicate significant pvalue in Tables

Indicate p values in Table 4

Gender should be separated from Age group in Table 4
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 INFLUENCE OF MATERNAL BODY MASS INDEX AND PHYSICAL ACTIVITY ON SELECT CARDIOVASCULAR RISK FACTORS OF PREADOLESCENT HISPANIC CHILDREN
Review round: 1
Reviewer: 2

Output:
Basic reporting: OBJECTIVE & COMMENT
Clear and unambiguous, professional English used throughout.
	English is clear and unambiguous and professional. Authors are urged to refrain from using multiple terms to describe one topic throughout (i.e. cardiovascular risk). See comments below.
The article must be written in English and must use clear, unambiguous, technically correct text. The article must conform to professional standards of courtesy and expression.
	English is used properly. Authors are urged to used concise statements throughout to improve reader understanding.
Literature references, sufficient field background/context provided.
	Literary references are sufficient but not optimal. See commentary within each recommendation.
The article should include sufficient introduction and background to demonstrate how the work fits into the broader field of knowledge. Relevant prior literature should be appropriately referenced.
	Sufficient introduction and background are provided. Authors are reminded that these sections should stay highly focused on the topic of interest and should not sway from the specific objective.
Professional article structure, figs, tables. Raw data shared.
	Figures are appropriate and legible. Recommendations regarding study model are made below. Primary raw data was not provided.
The structure of the article should conform to an acceptable format of ‘standard sections’ (see our Instructions for Authors for our suggested format). Significant departures in structure should be made only if they significantly improve clarity or conform to a discipline-specific custom.
	Standard sections are appropriate. The authors are urged to consider including a concise ‘conclusion’ to the abstract.
Figures should be relevant to the content of the article, of sufficient resolution, and appropriately described and labeled.	Figures are appropriate.
All appropriate raw data has been made available in accordance with our Data Sharing policy.	Raw data was not provided; authors provide appropriate explanation as to why.
Self-contained with relevant results to hypotheses.	Hypotheses are clearly stated but may need improved fluidity. See comments below.
The submission should be ‘self-contained,’ should represent an appropriate ‘unit of publication’ and should include all results relevant to the hypothesis.	The manuscript fulfills this requirement. Comments regarding hypothesis representation are made below.
Coherent bodies of work should not be inappropriately subdivided merely to increase publication count.	Although it is no required to remove, it is evident there are some unnecessary subdivisions (i.e. perceptions data).

The following statements are agreeable to the reviewer and should not be removed from the manuscript if possible:
Agreeable: Authors use appropriate source for recommended child screen time.
American Academy of Pediatrics is an excellent and appropriate source choice for establishing the recommended screen time for children. Authors appropriately describe how data falls into categorization based off these recommendations. See lines 147- 151.
Agreeable: Authors clearly define hypothesis to prepare readers for analysis.
Authors provide an excellent tool for communicating the purpose of the article. Further, the hypotheses are appropriately placed within the articles. Although hypothesis should not be changed after commencement or completion of the research, authors are encouraged to read statements on track changes document for suggestions on further study hypothesis. See lines 82-89.

The following are recommendations listed from most recommended to less recommended:
Recommendation: Change the title to reflect the findings. It is important to keep terms “Hispanic children” but “cardiometabolic risks” should be replaced.
The manuscript title should reflect the data findings and appropriate context. Consider replacing “cardiometabolic risks” with “healthy lifestyles” or “physical activity, blood pressure, and body mass index”. Cardiometabolic risks are not fully represented in this article. See the clinical application, diagnosis, and definitions for cardiometabolic risks. You may consider using the term “atherosclerotic risks”. The concern here is that the conclusions may be considered invalid if the cardiovascular risks and/or atherosclerotic risks have not been fully and properly being assessed. BMI may be used as a screening tool but cannot be used alone to classify a child for cardiometabolic risk entirely. Suggested titles include:
•	The influence of maternal BMI on physical activity of preadolescent Hispanic children
•	Hindrances to physical activity in Hispanic children
•	The relationship between maternal BMI and healthy lifestyles in preadolescent Hispanic children
Nichols GA, Horberg M, Koebnick C, Young DR, Waitzfelder B, Sherwood NE, et al. Cardiometabolic Risk Factors Among 1.3 Million Adults With Overweight or Obesity, but Not Diabetes, in 10 Geographically Diverse Regions of the United States, 2012–2013. Prev Chronic Dis 2017;14:160438. DOI: http://dx.doi.org/10.5888/pcd14.160438
US Endocrinology, 2007;(1):60-3 DOI: http://doi.org/10.17925/USE.2007.00.1.60
Wilson PW, Meigs JB. Cardiometabolic risk: a Framingham perspective. Int J Obes (Lond). 2008 May;32 Suppl 2:S17-20. doi: 10.1038/ijo.2008.30. Review. PubMed PMID: 18469835.
Kelly, A.S., S.E. Barlow, G. Rao,et al. 2013. Severe obesity inchildren and adolescents: identification, associated healthrisks, and treatment approaches: a scientific statement from the American Heart Association. Circulation 128: 1689–1712.
Flegal, K.M., R.Wei, C.L. Ogden,et al. 2009. Characterizing extreme values of body mass index-for-age by using the 2000 Centers for Disease Control and Prevention growth charts. Am. J. Clin. Nutr. 90: 1314–1320.
Recommendation: Terms ‘cardiovascular’, ‘cardiometabolic’, ‘obesogenic’ [, ‘metabolic syndrome’, and ‘atherosclerotic’] should not be used interchangeably.

The authors should pick a term and hone in on it. Try to refrain from introducing new terms into the article unless they are essential to the article. See title, lines 1, 24, 27, 53, table 1, 2.

Similarly, authors should be wary of terms ‘overweight’, ‘obese’, ‘severely obese’, and ‘BMI’. Consider using clinical vernacular to define these terms at the beginning of the article and choose one or two variables to assess throughout the course of the manuscript.

Another example of this is words ‘height’ and ‘weight’ being used concurrently with BMI. Define the variable and choose one.  That is, if authors decide to study BMI relationships, define how BMI is calculated and discuss BMI. Discussions and statements regarding ‘height’ and ‘weight’ are hence redundant and should not be included unless defining another variable of the study.

https://www.cdc.gov/obesity/adult/defining.html

Recommendation: Replace ‘children’ and ‘young children’ with ‘preadolescent’.
The authors target a very intriguing child age range. Authors should state specifically what range that is within the abstract and introduction. Nevertheless, age ranges of child subjects are disclosed analyzed appropriately.
Recommendation: Consider including best practices for the subject matter from credible sources in the introductory area of the article.
Best practices are essential for providing the reader with the appropriate knowledge and context. Authors may also consider the prevention strategies recommended by the Center for Disease Control and Prevention (https://www.cdc.gov/obesity/resources/strategies-guidelines.html). CDC statements on obesity are included but strategies, guidelines, and recommendations are not.
American Heart Association. (2013). Healthy Way to Grow. Retrieved from http://www.healthywaytogrow.org/HWTG/ and https://www.heart.org/idc/groups/heart-public/@wcm/@adv/documents/downloadable/ucm_473791.pdf
Kumanyika, S.K., et al. (2008). Population-based prevention of obesity: The need for comprehensive promotion of healthful eating, physical activity, and energy balance. Circulation, 118(4), 428-464. doi: 10.1161/circulationaha.108.189702.
Pate, R.R., Davis, M.G., Robinson, T.N., Stone, E.J., McKenzie, T.L., & Young, J.C. (2006). Promoting physical activity in children and youth a leadership role for schools: A scientific statement from the American Heart Association Council on Nutrition, Physical Activity, and Metabolism (Physical Activity Committee) in collaboration with the councils on Cardiovascular Disease in the Young and Cardiovascular Nursing. Circulation, 114(11), 1214- 1224

Recommendation: State the increase of obesity prevalence among Hispanics and/or entire US population.
Authors should consider presenting the relevance to this problem by providing a numerical value of increased incidence and prevalence over the past 10-20 years. Authors mention prevalence values, but this does not represent the increased incidence over a long period of time. Although not necessary, authors may consider providing values specific for Tennessee, US, non-Hispanic, Hispanic, age-specific onset and others.
Recommendation: ‘Discussion’ portion of the abstract should be edited.
Please see track changes document submitted. The discussion portion should either reflect a broad overview of the findings and a concise conclusion or only a broad overview of the findings. If the latter, authors should consider including a ‘Conclusion’ subsection to the abstract containing 40 letters or less.
Recommendations: Delineate from whom ‘negative perceptions’ are being assessed early in the manuscript.

Delineating this terminology is important for readers to understand early on. If not described early, the reader may remain uncertain whether the authors are discussing negative perceptions of community stakeholders, of the child subjects, of the mothers, of the parents, and so forth. Noteworthy, the assessment of community perceptions is complex and difficult to interpret. A recommendation to omit several, if not, all variables regarding maternal negative perceptions is expressed. See considerations below. The article is not suitable for submission to a survey research journal nor journals with a high degree of reliance on survey research methods.

The following are comments on considerations for the authors. No action necessary unless otherwise decided by authors:
Consideration: Include a ‘Purpose’ subsection within the abstract to state the scope of the article in 1 or 2 sentences.
The background and methods section may be perceived as discombobulated. There are several variables being discussed, some of which are not represented in the data. Consider making a short and concise statements such as “This study will analyze if maternal BMI, PA, and perceptions pose risks for adolescent BMI, PA, and BP in a Hispanic subject set”. Although this is described in depth in the abstract and other areas, it may be difficult for the reader to understand what the author’s goals are after reading the abstract. See track changes document lines 78-81 to see how purpose may be reworded in a clear and concise manner.
Consideration: Include a statement on inheritability, or absence thereto, of atherosclerotic disease and obesity.
Because the authors are claiming that there is existing research on obesogenic factors and their correlation to the health of older children, it may be appropriate to state whether genetics may play a role. If the influence of environment plays a greater role, authors should state so and why.
Consideration: Avoid the use of double negative and double/triple “less-more-than-most-at least” terminology.
Although such terminology must be used for statistical analysis and study structure sometimes, authors should strive to avoid it, if possible. Extensive use of double negative variables, analysis, and phrases lend to decreased clarity. Although this is not means for omission, nor is it incorrect, authors should be wary of such writing techniques. See lines 40-44, 216, 219-226, 227-233, 234-236.
Consideration: Stay mindful of citation style and requirements of the choice journal.
Experimental design: OBJECTIVE & COMMENT
Original primary research within Aims and Scope of the journal.
	Not applicable. Aims and scope not shared.
Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.	Sufficient.
The submission should clearly define the research question, which must be relevant and meaningful.	Sufficient.
The knowledge gap being investigated should be identified, and statements should be made as to how the study contributes to filling that gap.	Sufficient.
Rigorous investigation performed to a high technical & ethical standard.	Sufficient but needs improvement.
The investigation must have been conducted rigorously and to a high technical standard. The research must have been conducted in conformity with the prevailing ethical standards in the field.	Sufficient.
Methods described with sufficient detail & information to replicate.	Excellent. Consider condensing information in this section. Authors do an excellent work in describing the methods, especially inclusion and exclusion criteria.
Methods should be described with sufficient information to be reproducible by another investigator.	Sufficient.

The following statements are agreeable to the reviewer and should not be removed from the manuscript if possible:
Agreeable: Inclusion of age-based pediatric growth reference charts are an excellent tool used by the investigators.
The authors use the appropriate CDC Aged-based Pediatric Growth Reference Charts to characterize their subjects. This tool provides a quantitative value that can be used in a myriad of statistical analysis.
Agreeable: Raw data receives appropriate adjustments.
Authors report appropriate adjustments to raw data set characterizations such as maternal education and child age and gender.
Agreeable: Maternal blood pressure is not included in analysis.
Because maternal BP may be highly variable due to pharmacological use, pregnancy, or other, it’s understandable that they authors did not include this highly fluctuating variable as part of their analysis.
Agreeable: Statistical methods utilizes are acceptable.
The use of chi-squared, Fisher’s exact tests, for sociodemographic characterization of the data set and bivariate analysis is appropriate for this study. Noteworthy, authors should consider how categorization of quantitative variable values are represented in this statistical analysis.
Agreeable: Authors exclude patients with existing serious physical or mental illness.
Because authors state they’ve excluded patients with serious and mental illness, the reader can understand that subjects used for this study are less likely to have confounding biases and the like.

The following are recommendations listed from most recommended to less recommended:
Recommendation: Inclusion criteria should specify maternal ethnicity.
Whether or not all mothers identified as Hispanic, it is important to specify such to the reader. If subject dyads include non-Hispanic mothers to Hispanic preadolescents this renders a different conclusion than if subject dyads include only Hispanic mothers to Hispanic preadolescents. That is, it should be stated if there is a possible paternal factor involved. If the latter is how the data qualifies, consider using ‘Hispanic households’ to identify the parental subjects instead of ‘maternal’ or ‘mother’. If such is the case, households should be screened for single-parent households.
Recommendation: State the existing research.
Maternal obesity and lifestyles have been researched regarding their effects on infant and child atherosclerotic risk. Positive family history, and the presence of dyslipidemia in parents, children or adolescents has been previously investigated. One such example is given below. Although the article aims to discuss other items, it is important to inform the reader that maternal atherosclerotic factors have been associated with childhood atherosclerotic risk. If the authors decide to omit terminology such as ‘cardiometabolic’ and the like, this recommendation may be dismissed.
Napoli C, Glass CK, Witztum JL, Deutsch R, D'Armiento FP, Palinski W. Influence of maternal hypercholesterolaemia during pregnancy on progression of early atherosclerotic lesions in childhood: Fate of Early Lesions in Children (FELIC) Study. Lancet 354, 1234-1241 (1999).
Recommendation: Omit neighborhood ratings variable.
The authors do not report any perceptions below ‘somewhat good/average’ satisfaction and, therefore, should not be used to contribute to hypothesis number one. That is, if there are no subjects that report a negative perception, the data cannot be used to test for a correlation to child physical activity, BP, and BMI. See Table 2. Because the authors state in their first sentence that “negative perceptions of neighborhoods have been identified as obesogenic”, the authors should revisit those findings to create questionnaire that are in alignment with what is considered “negative perceptions”. Similar dilemmas -may be found in maternal BMI, see lines 175-177.
Recommendation: Condense confidentiality and informed consent statements.
Consider condensing this statement found in lines 109-115 to, for example, “Mother-child dyads were provided written and oral information on the study protocols and purpose and thereto were requested to provide written voluntary informed consent; all of which consented. Children seven years or older and mothers were required to provide written informed consent to proceed with the study.”

The following are comments on considerations for the authors. No action necessary unless otherwise decided by authors:
Consideration: If possible, include odds ratio values in the Table 3. Bivariate associations between children’s physical activity, TV viewing, blood pressure, and BMI.
Consideration: The authors may find it relevant to state that multiple logistic regression models were used specifically to calculate respective odds ratios.
Consideration: Authors may consider removing perceptions as a variable.
First, the perceptions measured are numerous; this quantity may supersede the necessary measures for communicating the intended analysis. It may cause confusion to the reader as to what the core variables studied are. Secondly, the significance, or insignificance, of maternal perceptions of their community are not stated within the conclusions. Because this was presented as a pinnacle of the study in the abstract and introductory statements, authors should consider including a concluding statement or removing the variable. Third, the questionnaires, although acceptable, are not structured to the maximum benefit to the study. Because the study has been completed, data cannot be modified but authors should be wary of how the survey methodology may result in unintended data results that do not reflect the purpose of the article. Lastly, there are three questionnaire variables that pertain to neighborhood perceptions. Authors can consider omitting all three, condensing these three, or omitting two of the three. This would allow the reader to assess Table 2 much more fluently. It is unclear why three perception variables are included
Consideration: TV screen time should be discussed further.
TV screen time is a progressively growing measure within the field of public health because of the increased use of screens in today’s children. This shows that the authors have a good understanding of the issue at hand. Although, data and the importance of TV screen time presented to the reader is only lightly discussed. Consider replacing ‘TV screen time’ to ‘screen time’ since authors used TV and video as a measure of screen time.
Validity of the findings: OBJECTIVE & COMMENT
Impact and novelty not assessed. Negative/inconclusive results accepted. Meaningful replication encouraged where rationale & benefit to literature is clearly stated.	Impact is stated clearly. Authors are urged to discuss why non-Hispanic groups were not used further and how future longitudinal studies including non-Hispanic groups would be beneficial.
Decisions are not made based on any subjective determination of impact, degree of advance, novelty, being of interest to only a niche audience, etc. Replication experiments are encouraged (provided the rationale for the replication, and how it adds value to the literature, is clearly described); however, we do not allow the ‘pointless’ repetition of well known, widely accepted results.	Sufficient.
The data on which the conclusions are based must be provided or made available in an acceptable discipline-specific repository. The data should be robust, statistically sound, and controlled.	Data not available to reader, authors provide appropriate explanation why and willingness to share upon specific request.
Conclusion are well stated, linked to original research question & limited to supporting results.	Conclusion needs modification to reflect pinnacle results of the data and hypothesis findings.
The conclusions should be appropriately stated, should be connected to the original question investigated, and should be limited to those supported by the results.	Conclusion is clearly stated but needs improvement. See comments below.

The following are recommendations listed from most recommended to less recommended:
Recommendation: Authors should state the purpose behind using categorical statistical analysis for quantitative values, such as BMI.
The readers should be well informed on why BMI ranges were used for statistical analysis instead of numerical BMI values. This could potentially be affecting statistical outcomes especially since four categories were grouped into two for analysis. See lines 136-138. Similar categorization is found for adolescent age groups one and two. See lines 184-186.
Recommendation: Authors should state how longitudinal studies remain necessary for further investigating the correlation between mentioned maternal factors and mentioned preadolescent factors.
The association between various maternal atherosclerotic predisposing factors, such as physical activity, and child body weight, particularly obesity, has been investigated. Although, sufficient longitudinal studies have yet to be published. This and other similar future studies should be mentioned in attempt to clarify the remaining gap of knowledge.
Recommendation: Briefly discuss the lack of multicultural analysis for this topic and how it would further contribute to the issue.
Recommendation: Future studies with different statistical analysis and inclusion of control groups.
It is important for the authors to recognize that, although thorough, the data cannot make assumptions regarding inclined risk of any of the mentioned variables in Hispanic children, as compared to their White non-Hispanic peers. Although past studies have established several of these correlations, this article does not provide sufficient controls to make such conclusions.

The following are comments on considerations for the authors. No action necessary unless otherwise decided by authors:
Consideration: Clarify whether the 118 subjects had or had not metabolic syndrome.
It appears the data that was used for this manuscript as extracted from a previous study. This is a useful method for clinical research, namely because existing data set may be difficult to obtain in select communities. Having said this, it is unclear whether the data set that was used in this study was extrapolated from an existing set of subjects for whom which all had metabolic syndrome. If so, it is important to state that subjects that reported pre-existing or diagnosed metabolic syndrome were included in this study and that no non-metabolic syndrome subjects were used in this study.
Consideration: Explain that this article is only representative of Hispanic adolescent physical activity over the past 7 days and TV screen time over the past 30 days.
Although the authors state that this is a cross-sectional data set, the reader only learns that the questionnaire data only represents maternal perceptions of 7-day and 30-day periods. For future studies, authors should consider including “on average” with relevant questions and refrain from ‘within the past X days’ to improve sampling.
Additional comments: Authors clearly state the lack of any conflicts of interests. Similarly, I, Gwendolyn A. Quintana, do not have any conflicts of interests to report.
Although not directly representative of this study here are some articles that may be of use for the authors for assessing the application of their study to the broader topics mentioned.
Cui, Z; Truesdale, K P; Bradshaw, P T; Cai, J; Stevens, J. International Journal of Obesity; London Vol. 39, Iss. 8,  (Aug 2015): 1203-1208. DOI:10.1038/ijo.2015.56
Oliveira, F. L. C., Patin, R. V., & Escrivão, M. A.,Meil Schimith. (2010). Atherosclerosis prevention and treatment in children and adolescents. Expert Review of Cardiovascular Therapy, 8(4), 513-28. doi:http://dx.doi.org/10.1586/erc.09.170
I would like to thank the authors for their contributions to maternal and adolescent public health through the work done in this manuscript and more to come. Please do not hesitate to request additional information or clarification regarding the review of this manuscript.


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

Input:
Title: THE INFLUENCE OF MATERNAL BODY MASS INDEX AND PHYSICAL ACTIVITY ON SELECT CARDIOVASCULAR RISK FACTORS OF PREADOLESCENT HISPANIC CHILDREN
Review round: 1
Reviewer: 3

Output:
Basic reporting: This paper is clear and with proper English used.
Literature references provided sufficient context to te work.
Experimental design: The experimental design is very well explained. This investigation has a rigorous methodology and complies with ethical standards.

I would only suggest explaining in a more detailed way "A trained research staff".
Further, please add the missing abbreviations in all tables (eg."BMI").
Validity of the findings: Although data is not so high, its statistics are sound and controlled, conclusions are fair within the results and, eventually, will hamper a future larger study.
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 INFLUENCE OF MATERNAL BODY MASS INDEX AND PHYSICAL ACTIVITY ON SELECT CARDIOVASCULAR RISK FACTORS OF PREADOLESCENT HISPANIC CHILDREN
Review round: 2
Reviewer: 1

Output:
Basic reporting: I do not have anything to add.
Experimental design: I do not have anything to add.
Validity of the findings: I do not have anything to add.
Additional comments: I do not have anything 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: THE IMPACT OF PNEUMOCOCCAL VACCINATION ON PNEUMONIA MORTALITY AMONG THE ELDERLY IN JAPAN: A DIFFERENCE-IN-DIFFERENCE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: This clearly written paper is on a very important and unsettled question. The analysis and the figures are clearly laid out and described. The authors describe the existing literature on the effectiveness of PPCV13 and PPV23 and place their paper in this literature.
Experimental design: The research question is well defined and the analysis is well described.
Validity of the findings: All my concerns have to do with the analysis. In order of importance.

1.	The authors should do more to assess the parallel trends assumption. They are finding very big effects that are not visible in Figure 1B. These may be the result of diverging trends. Any progress the authors make on directly assessing the parallel trends assumption such as presenting the differences in mortality rates between pneumonia and the control groups would strengthen the analysis.

2.	Why are the regressions specified to capture a break in the trend rather than a sharp change in the level? Is it due to the slow roll out of the vaccine?

3.	More detail on vaccine uptake would really strengthen the paper. This would allow the readers to scale the effects. If uptake is as low for the relevant population as suggested by the overall numbers in the paper, then treatment on the treated is much larger than the effects documented in the paper. Unless of course there are large herd effects.

4.	It may be worth implementing a difference in difference analysis that leverages the age specific recommendations for the vaccine. This might provide a cleaner comparison or at least one more counterfactual. People 60-64 might provide a better counterfactual to those 65-69 than the cause of death counterfactual.

5.	Regression specification (1) allows for a level change (P_t) at the time of the roll out of the second vaccine. This might absorb some of the treatment effect.
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 IMPACT OF PNEUMOCOCCAL VACCINATION ON PNEUMONIA MORTALITY AMONG THE ELDERLY IN JAPAN: A DIFFERENCE-IN-DIFFERENCE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall, the article was clear throughout and structured in an easy to read format.  The literature review seemed lacking as there are many articles on the effectiveness of the pneumococcal vaccine and not many are mentioned.   There are a few additions that I think would improve the paper:

1.  I would also like to see tables with basic summary statistics of the data.  I'd like to know the underlying rates of pneumonia mortality by gender and age to be able to have a reference point for the estimates that you find.  They seem like very large numbers (perhaps implausibly large... but it is hard to know without the summary statistics.)

2.  I would like to see tables of the results.  While I like the results figures, it would be nice to see the coefficients and standard errors in an easy to read table format that also includes whether standard errors are robust to heteroskedasticity or clustered.

3.  I would include the data from the observed values of figure 3 along with a line to mark the introduction of the pneumococcal vaccine to motivate the DID design.  I'm not sure that the predicted values add much to the graph or reporting of the results.
Experimental design: I agree that the rollout of vaccination allows for a nice difference in differences design.  I question whether the choice of heart disease and malignant neoplasms are appropriate control groups.  The authors defend this choice as these are prevalent causes of death.  However, it seems that something like pneumonia might have more seasonal and year to year variation than heart disease or malignant neoplasms.  I'd think that respiratory illness would be a more appropriate control group.

I am also not sure why the results only focus on the elderly given the vaccination of children beginning in 2013.  It seems that childhood mortality would also be an interesting outcome measure.  I am also not convinced that 2013 is the appropriate year to use for the post treatment variable with elderly outcomes since elderly vaccination did not occur until 2014.  It would at least be nice to see that the results are robust to using 2014 as the post year.

I am assuming that the data are only available in 5 year age bands.  If not, then age should be incorporated more fully in the DID design since different ages were vaccinated at different times.

Finally, it would be nice to show vaccination rates over time to provide support for the DID design.  Given some vaccinations started prior to 2014 and the vaccinations were phased in gradually after 2014, it is not clear that there really is a clear jump in vaccinations during the post period.
Validity of the findings: No further comments.  See above for some concerns about the methodology which would cause concerns for the 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: THE IMPACT OF PNEUMOCOCCAL VACCINATION ON PNEUMONIA MORTALITY AMONG THE ELDERLY IN JAPAN: A DIFFERENCE-IN-DIFFERENCE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Clearly written.
Experimental design: Well described and sensible
Validity of the findings: Look reasonable.
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 IMPACT OF PNEUMOCOCCAL VACCINATION ON PNEUMONIA MORTALITY AMONG THE ELDERLY IN JAPAN: A DIFFERENCE-IN-DIFFERENCE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: I appreciate the summary table of underlying mortality rates and seeing the results in table form.  I think this makes the paper easier to read and evaluate.  I also appreciate some data on vaccination rates, although more details across age and time would have been nicer.
Experimental design: I am still concerned that the control group is heart disease and malignant neoplasms.  The authors did not try to use another control such as respiratory disease… or perhaps deaths in another country that does not have a pneumococcal vaccination program (see Vivek Charu et al, 2011 for a cross country design).  Given the seasonal nature of pneumonia, a few years off trend from the more steady trend of malignant neoplasm is not surprising.
Validity of the findings: In light of reporting on the vaccination levels and lack of robustness with other control groups, I am concerned about the validity of the findings.

The vaccination levels suggest small uptake in the elderly which makes any direct effects seem unlikely (especially because the vaccine only protects against a fraction of pneumonia cases to begin with).  Moreover, vaccination rates of the children have not been given prior to 2013 to support the story that vaccination increased in 2013.  More details of the child policy need to be explained (which ages were vaccinated? And what do vaccination rates look like prior to the policy).    If we are to believe the results, this is really a story of a herd effect that gives protection to the elderly from the vaccination of youth.  The elderly vaccination rates changed very little suggesting that any effect can’t be driven by direct vaccination (unless the change in rates was significantly bigger for the 80+ age group than the numbers reported on the whole.)  Thus, I think the paper should be written more clearly to express this idea.

In addition, given that the effects must be driven by the youth, it seems that we should expect to see a jump in the data at 2013, not a slow decline in mortality from 2014.  It is not clear that we see that in the data.  Again, it would help to understand the child policy better and the evolution of vaccination rates among children to understand what we expect to happen in the data.
Additional comments: I believe that the results cannot reflect effects of elderly vaccination given small changes in vaccination rates and large effect sizes.  Thus, the paper needs to be written more clearly to reflect an argument of herd effects.  In addition, I think more evidence of vaccination rates among children across time and age need to be presented to make the argument for herd effects.


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 IMPACT OF PNEUMOCOCCAL VACCINATION ON PNEUMONIA MORTALITY AMONG THE ELDERLY IN JAPAN: A DIFFERENCE-IN-DIFFERENCE STUDY
Review round: 3
Reviewer: 1

Output:
Basic reporting: The paper was clear and easy to read.  No further comments.
Experimental design: No further comments.
Validity of the findings: I think the claims seem more reasonable in this version of the paper.
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: RANAVIRUSES AND REPTILES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Lines 391: This is an awkward fist sentence. I suggest eliminating it and beginning with the second sentence currently on line 392.

Line 45-46: I suggest changing “noticed” to “reported” and adding Duffus et al. (2015), “Distribution and Host Range of Ranaviruses” in Ranaviruses, as a citation.

Lines 176-177: This sentence may be better placed in introduction.

Lines 37-42: This section of the introduction seems simplistic. I think the brief description of non-avian reptiles is unnecessary. As the author’s note, most ranavirus research has been conducted on amphibians and the purpose of this manuscript is to summarize reptilian ranavirus research. In this light, I think the discussion on lines 85-90 would work better as part of the introduction.

Lines 351 – 353: See also: Echaubard et al., 2010; St- Amour et al., 2008; Haislip et al., 2012; Reeve et al., 2013, Forson and Storfer 2006; Kerby and Storfer 2009.
Experimental design: No Comment
Validity of the findings: Lines 321-324: Although some research had shown ranaviruses can persist and remain infective in, there are other investigations that indicate that ranaviruses degrade and lose infectivity rapidly in ecologically relevant conditions, perhaps limiting effective transmission to direct contact. Brunner et al. (2015) summarize this in “Ranavirus Ecology and Evolution: from Epidemiology to Extinction: in Ranaviruses.
Additional comments: Lines 203-230: Is it possible to add general patterns of susceptibility to this section or perhaps add a susceptibility own section? Brunner et al. (2015) state, “Until the recent studies by Brenes et al. (2014a ,b) comparative studies involving reptiles were generally lacking.” Has this changed in the three years since the publication of Ranaviruses? If certain clades are more susceptible, even at broad scales, it would be a valuable addition. Alternately, does ecology influence susceptibility; i.e., are aquatic and semiaquatic reptiles less susceptible than terrestrial reptiles due to evolutionary history with ranaviruses? Even if this information is sparse or unknown, it would be a valuable inclusion as a future research direction.

Lines 386-388: I recommend a sentence or two explaining a little more about this relationship, particularly the hypothesis that ranavirus may use macrophages as a route of infection. See Grayfer et al. (2015) “Ranavirus Host Immunity and Immune Evasion” in Ranaviruses.

Line 140-142: I recommend briefly describing the differences, pros, and cons, particularly between swabs and blood samples. It would be valuable for researchers designing a sampling method to have these comparisons in once place.


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: RANAVIRUSES AND REPTILES
Review round: 1
Reviewer: 2

Output:
Basic reporting: In a few cases, references are missing for specific statements, and a few additional references are suggested.
Experimental design: This is not an original primary research paper. It is a review of relevant literature with a specific focus.
Validity of the findings: As a review of literature, some of the standards do not apply. Based on the literature search, some of the conclusions are somewhat speculative.
Additional comments: Ranaviruses are important pathogens in a wide variety of ectothermic vertebrates. As the authors correctly note, work on these viruses in reptiles has lagged behind studies in amphibians. The authors provide an overview of recent literature on this topic as well as pointing out areas in which specific studies are still very much needed. While it provides an overview of the subject, the authors provide limited detail, and in some cases overinterpret the available data (or the lack thereof, e.g. when discussing the uses of antivirals and vaccines). The paper does not present novel information or research, but could be of interest to people embarking on studies in this field as well as a basis for discussion on future research projects. There are a number of specific points that need to be addressed before the manuscript can be considered for publication. The research aspect of the paper relies on a literature survey of the past decade comparing references to reptiles and amphibians in the ranavirus literature. Is there a reason that fish were left out of this survey? It would be interesting to see if the trend to more mentions of reptiles in titles and abstracts corresponds to a trend to more mentions of the broad host range of these viruses in general.
More specific comments on the text:
Taxonomy
Lines 68-70: This section does not reflect the full current taxonomy of the ranaviruses, which also still include Santee-Cooper ranavirus and Singapore grouper iridovirus, which to not group in the categories listed.
Line 71: “All of the viruses that infect reptiles”: As the authors have noted, this is an expanding field, it would be better to qualify this, e.g. found so far and described in the literature… The authors further state that ranaviruses found in reptiles all belong to the CMTV-like and FV3-like groups, however, cases of ATV-like virus infections in reptiles are mentioned in Duffus et al. 2015 (in those cases ECV was described).
Diagnostics and Surveillance
Lines 121-123: This section on antigen detection is unfortunately placed between two sections on antibody detection. Please rearrange the text to be more consistent.
Lines 125-133: This section remains very theoretical, while none of the actual surveys looking for antibodies against ranaviruses in reptiles are cited, e.g. Johnson et al., 2010 and Ariel et al., 2017. While these authors are cited further above, but the text never actually mentions what was found.
Lines 142-144: Be more specific. What samples? What tissues?
Lines 135-150: The authors should also discuss the various specific PCRs that have been used for ranavirus detection, as these can have different targets and different specificities (e.g. only FV3-like viruses vs. all ranaviruses).
Line 153: The authors refer to “these methods” but have not yet specified any methods.
Lines 159-163: Again, the authors could be more specific as to what has been detected and what information has been gained as suggested using these methods in reptiles.
Lines 165-174: The suggested methods have not yet been sufficiently critically discussed to evaluate how useful they would be in specific situations as suggested. What methods in particular are the authors suggesting, or are they suggesting the development of new methods, and if so, which ones and why?
Distribution and host range:
Line 176: While the distribution of ranaviruses around the globe range does theoretically overlap the geographic distribution of reptiles, this is not a helpful observation. For example, the knowledge of ranavirus distribution in Africa is almost non-existent, while the distribution of reptiles there is quite significant.
Line 180: The authors state that some reptile populations are naïve to these viruses. What populations would they not consider naïve and what evidence do they have to differentiate between naïve and non-naïve populations?
Lines 181-184: It is important to note that sampling protocol may strongly affect the results of such studies, and should be evaluated when drawing conclusions or making comparisons.
Distribution and host range:
Line 209: Do not use the word “population” when referring to a captive group. Also, the authors cite Alves de Matos et al. (2011) here, but that publication did study wild squamates. The text should be rewritten accordingly.
Line 227: The authors mention endothermy here, but it might be helpful to mention the role of temperature in virus growth in the introduction or taxonomy sections when talking about what classes of animals can be infected. This topic is also mentioned again at the end of the manuscript, but needs some context earlier for better understanding.
Pathology:
Lines 251-252: Please add references for these statements.
In general: Why are various transmission studies with reptiles (with one exception) not mentioned here?
Vectors
Possibly discuss this in conjunction with findings of virus in blood of infected animals as well as with transmission studies and the modes of transmission used.
Reservoirs:
Lines 321-330: See also Munro et al., 2016. Survival of frog virus 3 in freshwater and sediment from an English lake. J Wildl Dis 52:138-142. And: Johnson and Brunner, 2014. Persistence of an amphibian ranavirus in aquatic communities. Dis Aquat Org 111:129-138.
Treatment
In general, this section should be more cautious based on the lack of specific data. E.g. in the discussion of the use of antivirals, acyclovir was not shown to be effective against a ranavirus in vitro (Furguson et al., 2014 which is only available as a proceedings abstract under: http://members.arav.org/resource/resmgr/Files/Proceedings_2014/27.pdf, while Allender, 2012 discussed the possible use and pharmacokinetics, but did not demonstrate efficacy against ranaviruses.
Line 413 stating that „antiviral therapy is ineffective at reviving them“, possibly because animals are already in critical condition implies that an effective antiviral treatment is available, which is not currently the case. The same is true of line 417 „application of treatment….“
Lines 423-424: It might be more helpful to cite other examples of attempts to vaccinate reptiles against viruses, e.g. Jacobson et al. 1991 (rattlesnakes against paramyxoviruses, not successful), or cases in which crocodilians have been vaccinated against poxviruses and West Nile virus (see e.g. Horner, 1988). In this section it might also be helpful to mention possible carrier states in conjunction with vacicnation as a cautionary note.
Line 447: It would be helpful to include references for global declines of reptiles if this is mentioned as a driving force for continued research on this topic.
Figure 1: for interpretation of this graph, it would be helpful to note the number of publications taken into consideration again in the caption.
Table 1: I am confused as to how specific publications were chosen for this as it is not (by far) comprehensive. This needs either some explanation or needs to be significantly expanded.
Additional minor points:
When using taxonomical terms including family names in virology, these should be italicized (e.g. Iridoviridae).
The term Ranavirus (italicized, capitalized) refers to a taxonomic construct (genus), do not capitalize or italicize the word when referring to a specific virus.
When discussing the taxonomy of the hosts in which ranaviruses have been detected (in the abstract and the introduction), remember that lizards are not monophyletic.
Common names should not be italicized (see Burmese star tortoise, line 188)
The use of the term “turtle” vs. “tortoise” should be considered – in general, terrestrial turtles/tortoises are referred to as tortoises in American and British usage (see line 189 – Hermann’s turtles are usually referred to as tortoises in the USA and the UK).


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: RANAVIRUSES AND REPTILES
Review round: 1
Reviewer: 3

Output:
Basic reporting: This review article intends to summarize the research progress in relation with ranaviruses and reptiles. It seems likely that the article covers a wide range of aspects; but it really contains rather less information. For example, when you read the abstract, only the last sentence gives a bit information about what is presented in this article. In immunology related section of this article, only a few AMPs and IFN-gamma are mentioned, with the inclusion of only two Ig isotypes. In fact, there are quite a few other reports on immune system of reptiles, including turtle and lizard, and it is not corret to say that there only two types of Igs in reptiles.
Experimental design: It might be important to summarize various aspects in relation with ranavirues and reptiles.
Validity of the findings: Research progress is not well or deeply reflected in the article.
Additional comments: A detailed digging of data may be required to reflect the research progress in relation with reptiles and ranaviruse. There are more published articles to be cited.


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: RANAVIRUSES AND REPTILES
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The author's have addressed my initial comments and generally improved the overall quality of the manuscript. I believe it could be a valuable overview for researchers beginning investigations on ranavirus infections in reptiles.


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: RANAVIRUSES AND REPTILES
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript is much improved. It currently contains multiple typos and careless mistakes and should be carefully edited before final publication.
Experimental design: As previously stated
Validity of the findings: Much improved.
Additional comments: Specific comments:
Abstract:
Why did you remove “known to be” from the sentence “More recently, reports of ranaviral infections of reptiles are increasing with over 12 families of reptiles currently known to be susceptible to ranaviruses, most of which are turtles and lizards.ranaviral infection”?

Diagnostics and surveillance
Page 5, paragraph IHC, last line: “indicating that viraemia proceeded the systemic infection observed in these animals.” Should be preceeded.

Distribution, host range and impact
- Testudines (turtles, tortoises and terrapins)
Johnson et al. 2007 also infected red eared sliders.
Trionyx sinensis is currently known as Pelodiscus sinensis

Distribution and host range - Squamata (lizards and snakes)
Final line . ranaviruses in wild squamates: add a reference to Alves de Matos et al., 2011 here.

Pathology:
Line: “challenge experiments, and the fact the reports are often confounded is co-infections”: This line needs some work I am not sure what the authors are trying to say here

Reservoirs
and soil (even others replies.....: should be even other reptiles

Immunology - Innate
work in reptile hosts if limited: is limited
Ranavirus overcomes the antiviral defenses of macrophages and uses the cells: should be Ranaviruses overcome….and use…

Immunology - adaptive
Ranaivurs IgY: Typo, and these are not ranavirus antibodies, but antibodies against ranaviruses

Treatment
De Veo: should be written DeVoe (also in references)
Further investigation is required to determine the optimal temperature for increasing survival from aof ranaviral infectioninfected reptiles.: It would be good to mention here that this will likely depend on the host speices.

Table:
What does H stand for?


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: RANAVIRUSES AND REPTILES
Review round: 2
Reviewer: 3

Output:
Basic reporting: My comments have been met, and I do not have further comments.
Experimental design: N/A
Validity of the findings: N/A
Additional comments: no further 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: SHORT-TERM INTERVAL AEROBIC EXERCISE TRAINING DOES NOT IMPROVE MEMORY FUNCTIONING IN RELAPSING-REMITTING MULTIPLE SCLEROSIS—A RANDOMIZED CONTROLLED TRIAL
Review round: 1
Reviewer: 1

Output:
Basic reporting: Major issues-
Introduction-
1.	The Intro should be reorganized and broadened. The authors may state the hallmarks of pathology of MS with one paragraph, followed by the potential role of exercise in attenuating cognitive impairments in RRMS (or MS), then state why you chose interval exercise, not continuous aerobic exercise, as an intervention.
2.	Please specify why you chose to focus on memory in your manuscript. Accumulating evidences have supported that memory impairment is one of the core deficit in MS.
Methods-
1.	Line 96-97: If the authors recruited participants from only one hospital, how the effect of selection bias mitigated in this study?
2.	Line 112: how the authors quantify ‘severe’ cognitive impairment? Did the participants tested with their general cognitive ability, education level, socioeconomic status, and BMI (body fat %)? These factors are all closely related to cognitive function (or impairment).
3.	Intervention: More details are needed with regards to the content of intervention. For example, when you implemented interval training, please specify how long was the between-set recovery? The exercise:rest ratio is crucial to the effectiveness of interval exercise training, therefore should be clearly stated in the manuscript. Also, please specify the intensity of the exercise (e.g., % maximal aerobic power). Please also specify: How many participants administered the intervention at the same time? How many participants wore a HR monitor in each session? How many physiotherapists supervised each session?
4.	Did participants monitored and controlled regarding to their daily activities, such as reading and/or physical activity?
5.	More details should be provided regarding outcome measures, including cognitive functions, motor function & walking ability, and patient-reported outcome measures. Please also justify your decision of task/assessment selection. Were the cognitive tasks or fitness assessments you used valid in MS? Currently, the format is too rough.
Results
1.	Please provide statistical results, including mean and SD, p value, effect size, 95% CI) in the sections pertaining to effects of exercise on cognition, motor function and walking ability, fitness, and patient-reported outcomes.
Discussion
1.	The reviewer believes that several potential limitations should be subjected to the current study. For example, as mentioned in previous comment, the authors should elaborate the potential confounding effects from several background variables, such as general cognitive ability, education level, socioeconomic status, and BMI, and how the confounding effects can be mitigated in the current study. Also, did selection bias exist in the current study? If yes, please elaborate this issue (and how it can be mitigated).

Specific comments-
1.	Title- The reviewer suggests the authors to revise the term ‘aerobic exercise’. A more precise term such as ‘interval aerobic exercise training’ could be more appropriate as the term ‘aerobic exercise’ could refer to both continuous and interval exercise.
2.	Line 26, 34, and 36: Please give the full name for ‘MS’, ‘RRMS, and ‘EDSS’ for first appearances.
3.	Line 27: Please at least report the number of participants in each group and their mean and SD of age here.
4.	Line 83-84: Likewise, please at least provide the number of participants as well as mean and SD of age in the IG and CG, respectively.
5.	Line 108: Please specify what EDSS is for first appearance. It is possible that some readers are not familiar with the purpose and interpretation of this assessment.
6.	Line 110: Did the authors collect data on how long they have had taken the immunotherapy? While some participants may have longer history of immunotherapy than others, would this inter-individual difference affect your results?
7.	Line 114: Were participants’ readiness to physical activity confirmed by valid assessment, such as the Physical Activity Readiness Questionnaire (PARQ)?
8.	Line 117: What software was used to calculate sample size? G*Power?
9.	Line 121: If a two-tailed two-sample (independent) t-test was used for sample size calculation, please specify what metric (e.g., Cohen’s d) was adopted for detecting group difference.
10.	Line 121: Why the authors chose an alpha of .025 and power of .90, rather than .05 and .80? This should be clarified.
11.	Line 118-121: the authors stated several figures for detecting group difference (e.g., -5.8 pts on the VLMT learning subtest, -2.6 pts on the VLMT delayed recall test). Please justify the use of these figures or cite papers to support your decision.
12.	Line 132: please justify the choice of a 12-week intervention.
13.	Line 197-198: Please clearly elaborate the ANCOVA (e.g., 2 (group: IG, CG) x 3 (time: T0, T1, T2)).
14.	Results- Did participants included in the ITT and PP differed in terms of baseline data??
15.	Line 269: Should be ‘…less disabled, physically and cognitively, thereby…’
16.	Line 274: the sentence is incomplete here.
17.	Line 320: the sentence here is unclear to the reviewer.
18.	Table 1, 2, and 3: The tables should be refined. For instance, please make sure all the value presented aligned to either the left or right side of each column. Also, the reviewer cannot find the T2 data in either Table 2 or Table 3.
Experimental design: no commment
Validity of the findings: no comment
Additional comments: The present study addresses a relevant topic and has the potential to contribute to the literature and treatment of multiple sclerosis. Novelties of the current manuscript include the use of interval training, the employment of a comprehensive measure on multiple cognitive domains that are vulnerable to the progression of multiple sclerosis, and the follow-up assessments. Key findings of the current study are that, in individual with relapsing-remitting multiple sclerosis (RRMS), 8 weeks of interval training has no benefit to cognitive performance, including memory, executive function, and processing speed. Generally speaking, the manuscript is well-written, and the study was conducted in a professional manner. The authors should be credited by their efforts. Reservation, however, should be given to several parts of the study. The reviewer’s major concerns are to the methodological ends, with several issues worth clarification, justification, or contemplation. Please find the comments attached.


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: SHORT-TERM INTERVAL AEROBIC EXERCISE TRAINING DOES NOT IMPROVE MEMORY FUNCTIONING IN RELAPSING-REMITTING MULTIPLE SCLEROSIS—A RANDOMIZED CONTROLLED TRIAL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall the paper is well-written and tables and raw data have been provided.
Experimental design: This study reports the results an RCT involving 69 people with RRMS examining the effects of AE on memory. The topic is a timely one and even though these are ‘negative’ findings’, the report of the design and results are important for future studies. Participants in the AE group trained on bicycle ergometry 2-3 times per week for 12 weeks. Importantly, outcomes were examined at 12 week follow-up which has been a weakness of previous trials.
Validity of the findings: Good RCT practices were followed, for example the trial was registered and the authors ensured random allocation and blinding of assessors. The authors included multiple outcomes making the results more convincing.
Additional comments: For the most part, the limitations and design flaws are addressed adequately. For example, the study was aimed at cognition but the participants were not asked whether they in fact had cognitive problems. It was not surprising then that there was a ceiling effect. This issue was mentioned in the Discussion section.
I have two major issues involving the intensity and duration of the AE intervention that participants received.
First of all, the maximum sessions that participants could have received was 30 but in fact participants attended on average 19.5 sessions (if 3X week this would be about 4 weeks). The intervention was 2-3 times per week but which was it- 2 or 3? The difference between 24 sessions and 30 session is substantial. The authors should discuss the appropriate length of treatment to gain an effect of AE gleaned from other studies in healthy individuals and in people with neurological disorders such as stroke. I would suggest based on the literature, that 8 weeks duration or about 24 sessions would be optimal. The issue of suboptimal treatment duration is not adequately discussed. Please compare and contrast short duration (<8weeks) with longer duration (>12 weeks) in order to provide context for these negative findings. There are multiple studies examining the effects of AE on cognition in stroke for example (Ploughman and Kelly 2016 PMID:27661010) In Fig 1 consort diagram, although not specifically indicated, it seems that 11 people were excluded from PP analysis because they attended <18 sessions. This is not quite clear-please indicate how many people were excluded due to this reason. Furthermore, why did these people not attend? Non-compliance issues are important for the planning of future studies. Please indicate reasons.
The second issue with the intervention is the low intensity nature. In the AERCONN protocol the exercise is anchored by W recorded at Borg RPE 11/20. This seem to be very low; Borg 11/20 is considered ‘light’ (https://www.cdc.gov/physicalactivity/basics/measuring/exertion.htm) and current guidelines support that training should be in the ‘somewhat hard’ category to achieve moderate level (according to the MS guidelines http://www.csep.ca/CMFiles/Guidelines/specialpops/CSEP_MS_PAGuidelines_adults_en.pdf). According to the AERCONN protocol, the first 20 sessions (the majority obtained by most people), the training was in the ‘light’ or ‘mild’ training threshold. This is a major issue that has not been adequately addressed in the Discussion section. Please review the intensity of exercise required to change cognition in animal models and clinical trials in order to place the work in context. Furthermore, the intensity of the intervention should be clearly indicated in the Methods section.
The title and abstract should be edited to reflect that the intervention was not in fact ‘Short term aerobic exercise’; but was ‘Low intensity aerobic exercise’
Other issues:
There is Responder group analysis but it was not clear to me how ‘responders’ were identified. The term ‘responder’ suggest that the group improved in the primary outcome but in Table 7 there were no significant changes in the cognitive outcomes. In order to better understand responder outcomes it would help if there was further analysis. For example, was there a significant relationship between attendance and change in memory score (post-pre)? Or a relationship between improvement in fitness and cognitive score change?
There was 12 week follow-up but I could not find any mention of the results of the follow-up.
Where are the F values and effect sizes for the ANCOVAs?