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

Input:
Title: DEVELOPMENT AND EVALUATION OF THE PARENTING TO REDUCE CHILD ANXIETY AND DEPRESSION SCALE (PARCADS): ASSESSMENT OF PARENTAL CONCORDANCE WITH GUIDELINES FOR THE PREVENTION OF CHILD ANXIETY AND DEPRESSION
Review round: 1
Reviewer: 1

Output:
Basic reporting: First, I would really like to applaud the author for this work. I believe the work presented here will be very useful to many researchers working with the prevention of mental illness in children.

In terms of basic reporting, I believe the authors use a clear and professional English throughout the manuscript, the structure conforms to standards, tables are clearly presented and informative, and two data-files are supplied.

There are a couple of things that I think should be considered before the manuscript is being accepted.

1. In the introduction (lines 88-90), the authors write “To our knowledge, no current parenting measure exists that assesses the full range of parenting risk and protective factors that are known to influence the development of anxiety and depression in children”. I agree with the authors, however, it might be worth mentioning that there are questionnaires of parental behaviors highly relevant to the development of anxiety in children (although not “full-ranging”). E.g., The parental overprotection scale (e.g., Clarke, Cooper & Creswell, 2013) and Rearing behaviors questionnaire (e.g., Bögels & Melick, 2004). I suggest the authors mention these measures and briefly discuss in what way the paRCADS add to these existing measures.

2. The hypotheses presented in the Introduction (lines 130-135) seems somewhat arbitrary. I suggest that the authors present more background (or rational) of the expected directions and strengths presented here.
Experimental design: I believe the research is within the scope of the journal, that the research question is adequate, and that the manuscript involves a methodologically comprehensive approach.

There is one thing that could be improved, which I believe would strengthen the interpretation of the results later on:

1. Regarding recruiting participants (lines 150-158), I think it could be more clearly described what population the intervention (web-based parental program) was targeted to. This is important, as later in the manuscript it is obvious that this is not a “normal” population. For example, about 2/3 of parents had previous mental health disorders. Already at this stage in the manuscript it would be clarifying to describe the sample better.
Validity of the findings: From what I can see, data seems to be robust and adequately handled. And, I believe conclusions are generally well stated.

However, there are a few things that I would like to comment, that I think may be important to consider before accepting the manuscript.

1. Two things important to consider when exploring convergent (and discriminant) validity of a parent self-report measure is (1) the common-method bias (or common method variance) and (2) the general discrepancies of parent-child ratings. Common method variance means that the correlations between different parent-measures may be inflated. And contrary, the general discrepancies of parent-child ratings mean that the correlations between a parent-rated and child-rated questionnaire is expected to be weak. These commonly reported phenomenon could be good to highlight in order to understand why the authors specified “We hypothesised that convergent validity … would be demonstrated by: a) moderate to strong correlations with other parent reports of parenting; b) small correlations with child reports of parenting”. Further, highlighting these phenomenon may also help the reader to interpret the result section, e.g., why the child-rated parenting only showed a small correlation to paRCADS.

2. I agree that a weaker correlation between paRCADS and parental QoL compared to paRCADS and other measures of parenting support discriminant validity (i.e., evidens that the questionnaire measure aspects of parenting behaviors rather than parental QoL). However, also interesting is another level of discriminant validity, namely, whether the paRCADS really measures parenting behaviors related to risk/protective factors of anxiety/depression (as stated) or risk/protective factors of mental health in general.

Looking at the numbers (in Table 7), it seems that there is a stronger (positive) correlation between paRCADS and psychological well-being compared to the correlation (negative) between paRCADS and child anxiety/depression. This is true for both parent-ratings and child-ratings.

Perhaps the authors could discuss these findings. Could the paRCADS really be viewed as measuring parenting related to depression/anxiety? Or should it be defined to measure parenting related to mental health more broadly?

3. In the discussion section, the authors discuss the percentage of parents that were in concordance of the criteria. I believe this discussion would benefit from more clearly considering the specific sample examined in this study. Basically, it could be more clearly stated that this discussion concerns a specific group of parents (i.e., 2/3 with a prior mental disorder).

For example, the authors are concerned with the small number of parents that used adequate parenting regarding “managing emotions”. However, given this group of parents, this may not be so surprising, also, this result may not be replicated in a population-based sample.
Additional comments: Some minor issues:

1. They study involves a rather small age-range (8-11 years), perhaps add to limitations. Or propose to explore wider age-range in future research.

2. Define T-score (line 285), it may not be common knowledge what a T-score is (i.e., Mean 50, SD=10).

3. This sentence (line 337-338) is ambiguous “Little’s MCAR test suggested that item-level missing data can be reasonably assumed to be missing completely at random…”. Perhaps provide statistic details.


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

Input:
Title: DEVELOPMENT AND EVALUATION OF THE PARENTING TO REDUCE CHILD ANXIETY AND DEPRESSION SCALE (PARCADS): ASSESSMENT OF PARENTAL CONCORDANCE WITH GUIDELINES FOR THE PREVENTION OF CHILD ANXIETY AND DEPRESSION
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1)	I found this paper used the clear and unambiguous English throughout the paper. However I think that too much detailed description/information (which I believe is best for replication) in Materials and Methods & Results sections makes it difficult for readers to follow: for example, the specific values from the supplementary tables (e.g., line 544-546) authors mentioned in results section. Please find the excessively described writings and rephrase them in short throughout the manuscript.
2)	I think it is the same name/concept as the number of questions from Table 2 and the highest possible score from Table 3. Furthermore, the numbers of questions which are different in Table 2 and Table 3 makes readers feel confused how many items the subscale of PaRCADS consists of. I suggest you would delete Table 2 and add the example items from Table 2 into the text.
3)	I suppose if the Table 5 were presented as landscape view not as portrait view like Table 6, it would look nicer.
4)	Raw data has not been provided in the supplementary files. I suggest that you upload the raw data or explain the reason if you can’t.
5)	I can say authors conform the PeerJ standards very well in References section except the line 767: italicize ‘Educational and’ because it is the title of the Journal.
Experimental design: 1)	I find this study would contribute very necessary guidelines for the prevention and intervention of the mental health science based on the methodological soundness. However, authors mentioned relatively unfamiliar names of statistical methods such as point-biserial correlation, Little’s MCAR test, B-index, classification-consistency estimate, etc., so that some readers might not know what they are. Please explain the concept of them in short.
2)	Study 2 Expert ratings seems important for item selection using item content validity index (line 421-433). However, the 79 items were selected as the final total items though the study 2 reported that 76 items were endorsed by most experts. In addition, the study 2 repeatedly mentioned the file S2 and specific values again so that it might make readers puzzled to understand. Please find alternative ways to describe them (e.g., summarize them briefly not in separate section).
Validity of the findings: I understand the internalizing disorders are related to parenting factors such as overprotection and rejection according to the cited articles published by McLeod, Weisz & Wood (2007) and by McLeod, Wood & Weisz (2007). However, parental rejection and overprotection is associated with externalizing problems (e.g., Hale III, W. W., Van Der Valk, I., Engels, R., & Meeus, W. (2005). Does perceived parental rejection make adolescents sad and mad? The association of perceived parental rejection with adolescent depression and aggression. Journal of Adolescent Health, 36(6), 466-474; Buschgens, C. J., Van Aken, M. A., Swinkels, S. H., Ormel, J., Verhulst, F. C., & Buitelaar, J. K. (2010). Externalizing behaviors in preadolescents: familial risk to externalizing behaviors and perceived parenting styles. European child & adolescent psychiatry, 19(7), 567-575.). Considering that the latter article published by Buschgens and colleagues (2010) are utilizing longitudinal data, it is very helpful to understand the risk and protective parenting factors. Therefore I suggest you’d better add the possibility of the study that PaRCADS is good to deal with both internalizing and externalizing disorders.
Additional comments: I agree that the PaRCADS is valuable instrument to evaluate and intervene the parenting in research and clinical practices. If the title of the manuscript is changed into validation or development of PaRCADS: ~, it would be much easier for readers to grasp the contents of the paper.


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

Input:
Title: IDENTIFICATION OF MICRORNA SIGNATURES IN UMBILICAL CORD BLOOD ASSOCIATED WITH MATERNAL CHARACTERISTICS
Review round: 1
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: The manuscript by Juracek et al., “Identification of microRNA signatures in umbilical cord blood associated with maternal characteristics” demonstrated the UCB miRNAs could serve as biomarkers for pregnancy-associated complications. The content of this study is of greater interest and suitable for publication in the PeerJ. I recommend the authors could discuss some studies that show the application of pregnancy-related microRNAs in diagnosis or screening.


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

Input:
Title: IDENTIFICATION OF MICRORNA SIGNATURES IN UMBILICAL CORD BLOOD ASSOCIATED WITH MATERNAL CHARACTERISTICS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1) The report is well written, without language mistakes, structure of the report is standard, according to the journal requirements.
2) Background is clear and on the subject, allowing to understand the results and discussion and raw data are shared.
Experimental design: 1) The research described is original, performed by the standard and accepted methodology in the field.
2) In the material and methods section I lack the statistical evaluation and I suggest that you describe how were all the p-values obtained (tests etc).
Validity of the findings: 1) The research is interesting and fills a knowledge gap in the field.
2) The statistics should be described more in detail.
Additional comments: Well written and interesting report that paves the way for future possible miRNAs use in the diagnosis or prognosis of complications in obstetrics.


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

Input:
Title: IDENTIFICATION OF MICRORNA SIGNATURES IN UMBILICAL CORD BLOOD ASSOCIATED WITH MATERNAL CHARACTERISTICS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: 1. The miR array ( V4.0 microRNA Ready-to-use PCR Human Panel I+II TaqMan microarrays) was not found in Qiagen web, please check the information of commercial product was correct.
2. The requirement of include and exclude patients need to clarify.
Validity of the findings: 1. The fold change +_ 95% CI of up-regulated or down-regulated microRNAs should be listed in table 2.
2. Were there any references supporting the microRNAs change profile in the 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: IDENTIFICATION OF MICRORNA SIGNATURES IN UMBILICAL CORD BLOOD ASSOCIATED WITH MATERNAL CHARACTERISTICS
Review round: 1
Reviewer: 4

Output:
Basic reporting: Abstract:
Line 25: try 'useful' in place of ideal;
Line 30: CELSPAC: TNG study? – Provide explanation to acronym
Line 35: Results could be more specific, indicate miRNA pattern for deregulation (up or down?) and statistical outcome.
Line 36: we described A specific signature of - insert 'a'; Line 40: with a wide

Introduction:
Line 47: is blood that REMAINS in the placenta…
Lines 49-53: Could use references, e.g. Burns, E., 2014, 10.1891/1058-1243.23.1.41
 Authors Mithal et al (referenced on line 54) seem to refer only to the use of cord blood to predict sepsis.
Line 71: Moreover, PREVIOUS research has shown that …
Line 81: follow-up to? – Typo? Try 'which WILL follow up ON 10 000 children…'

Methods – Sample collection and processing:
We question the sample collection – how was the digestion of miRNA avoided, when samples were allowed to sit for 30 minutes. It seems there was almost an hour at room temperature for RNA to degrade. Contrary to this, the Qiagen’s kit used in this study does instruct sample processing as is presented in this manuscript. To better convince the reader, perhaps other sources could be referenced here, such as: Neyro, V., Elie, V., Médard, Y. and Jacqz‐Aigrain, E., 2018. mRNA expression of drug metabolism enzymes and transporter genes at birth using human umbilical cord blood. Fundamental & Clinical Pharmacology.
Line 89: The current study included 24 mothers from whom we collected umbilical …
Line 96: …and aliquoted into tubes as 250 μL samples and …

Methods – miRNA quantification:
Line 103-104: Could add catalog number, search does not bring up the product!
More information about normalization would strengthen this report, was Livak et al. method used: Livak, K.J. and Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), pp.402-408?
Were spike values taken into account when normalized, as suggested by Chen et al.: Chen, K., Hu, Z., Xia, Z., Zhao, D., Li, W. and Tyler, J.K., 2016. The overlooked fact: fundamental need for spike-in control for virtually all genome-wide analyses. Molecular and Cellular Biology, 36(5), pp.662-667?
Also please add details on statistical analysis following correlation with maternal factors: how was significance calculated? On that end, how was correlation performed?
Line 108: 50% not 50 %
Experimental design: Results:
Reviewer feels it would strengthen manuscript greatly if a scientific basis were delineated for these innate concepts: global profiling > statistical analysis > signature .  What do these terms really mean? Pattern? Trend? association? causal? Mechanism? In such a narrow scoped scan of 24 plasma samples (technical replicates?) can we talk about a 'signature'? Please define signature mathematically with probability of it meaning something clinically.

Throughout: make number presentation consistent, e.g. write out numbers one to nine, and present numbers over 10 numerically.
Line 112-113: Did all 24 samples contain each of these 752 miRs? Or was the kit capable of recognizing a total of 752 potential species? Please clarify this important distinction.
Line 117: associated with maternal age NAMELY miR−137… ?  Check sense, as 'thereof' is not clear in this context.
Validity of the findings: Discussion:
Line 154: Please clarify following sentence: ‘Despite negative results, this study delineates role of miRNA within delivery course and success.’
I would also suggest making it longer and putting results in the context of known literature, e.g. this would be a good spot to highlight some of the significantly correlated miRNAs and cite relevant functional studies, e.g. the best of the 333 'plasma papers' that cite Weber et al 2010 in Clinical Chemistry.
Line 154: Despite negative results - do you mean previous publications, or your Table 2? If the latter, then specify which entries are positive and which ones are negative.  Since no Hypothesis was delineated at the outset of the Experimental Design, then how can there be a "negative result"?
Line 156: "we expected that possible deregulation should be influenced by…" - Finally, this phrase is hinting at an a priori Hypothesis. Trouble, pain, questions, confusion all may lead to a Problem that is normally described in Background Literature Review. After the Problem is addressed then scientific inquiry proceeds to a Rationale and Hypothesis > Objectives >> Specific Aims... Placing the "expected possibilities" at the beginning of the Introduction would help Reader understand the potential usefulness of end-of-pregnancy plasma profiles.

Conclusions:
Line 161: "levels in … plasma are related to … characteristics" To Reader this statement appears to be an indication of POSITIVE results so it flies in the face of Line 154's "negative results". Please clarify.
162: Over interpretation? Have any of the miRNAs that were found deregulated been investigated in relation to maternal health, pathologies or complications?

Overall:
We think this study could offer a good resource for future research. We appreciate the supplementary table with raw reads, which might prove useful to other groups. We might suggest adding to a repository (e.g. GEO) for easier access. Literature search of miRNA and cord blood indicates that not many labs have explored cord blood’s general correlations to maternal health in the context of miRNA. This research would be a good addition to the community if: 1) data is uploaded to a repository; and 2) correlation and statistical analyses are presented in a more transparent and explanatory way.
Additional comments: Micro RNA means shorter than messenger RNA or transfer RNA, but is does not follow that another report on microRNA diversity of blood has to be 'micro' in manuscript length. The fact that the Introduction has just ~390 words implies that there is not much to report, or otherwise the subject matter of the experiment conducted is so self-evident or banal that it is not worth much of an overture. Results reported on Lines 120-134 state that a total of 74 miRs appeared changed in the two dozen samples of cord blood. How do these 74 species compare to miRs detected in other body fluids? Perhaps that question has no 'statistical power' for the approximately 2000 miR genes in the human genome of ~18k other genes? The repertoire of 'array hits' could be on a genomic-scale yet the Introduction does not present the experiment in a Hypothesis-driven approach. A shotgun snapshot strategy makes this random survey look too much like a fishing expedition, an observational 'systems biology' scan of a non-hypothesis small sample of newborn infants.
Weber et al's landmark 2010 report in Clin Chem has amassed over 1500 citations (Scholar), 1032 times cited in Clarivate's Web of Science. Given this plethora of attention one would expect a 2019 report to devote more than one line (Line 59-61) to miR biomarkers, especially given the fact that Weber et al. examined the presence of miRNAs in 12 human body fluids and urine samples from five (n=5) women in different stages of pregnancy. Their Table 3 lists 14 miRNA species that were uniquely detected in plasma so there should be ample opportunity to discuss this present manuscript in light of Weber and subsequent findings. How many hits from Juracek et al's Table 2 occur in the original 2010 report? Of these citers of Weber, 330 deal with plasma, 45 of these in the past twelve months, so there is no paucity of papers with which to compare these present UCB results from Juracek.
Reviewer has heard of an expression, the least publishable unit (LPU), and this manuscript coming in at Methods 365 words, Results 340, Discussion 195 should not be competing for such an accolade. If CELSPAC is in the same league as CALIPER (CIHR: Closing the Gaps in Pediatric Reference Intervals) then each of its publications should be broad, thorough and as deep as possible. Expand and cover the foundational literature on plasma miR profiling in clinical settings. Blood is blood. e.g. But these three smokers' compare how with the international profiles of smoker plasma profiles? Might you try to mine your data more deeply and meaningfully?


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: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English expression need to be further polished, and abbreviations should be completely spelt out when they are first appeared, such as "HOA" and "LOA" in line 30.
Experimental design: 1. The current study is a retrospective study and involved small numbers of people, which may make it less reliable.
2. According to the manuscript, during the one-year follow up, patients worn OK lens at night and removed them before 7 am on the next day (line 74). But authors didn't mention when patients worn OK lens (specific timepoint), so it's not sure if all patients worn OK lens in a fixed duration.
3. For optical quality measurement, pecific measure time wasn't mentioned in methods. Considering OK lens' influence to cornea would tail off with time, the measure time should be fixed to insure its veracity.
Validity of the findings: According to the result 3.4.1, astigmatism aberration had no statistical significance at 1 and 3 months, and was remarkablely higher at 12 months after wearing OK lens (line 149-151). But authors didn't explain this phenomenon or give possible mechanism in discussion, which may confuse the readers.
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: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. Please enhance the English writing of this study.  There were many grammar errors.
2. Age of "10.806±2.184" years old was not an appropriate format for age.
Experimental design: 1. Please describe the selection criteria in more details.  What eye examinations were done?
2. The "parameters were analyzed before and at 1, 3 and 12 months". However, in Table 1, only data from baseline and 12 months were presented.
3. Please detail the statistical methods used in analyzing the repeated data.  What type of student t-test was used?  Since Friedman test was a kind of repeated measurement ANOVA, what another ANOVA was used? How was the repeated measurement ANOVA applied to the data?  Did the authors checked the distribution of visual acuity and compensating factor?
4. How was the visual acuity measured?
5. As the orthokeratology lenses can pose potentially vision-threatening risk to the subjects, did the authors record any of the adverse events?
6. It is very important to know how the study subjects were selected.
7. Please define the error bars in the figures.
8. The authors should describe the compliance data during the study period.  Any other treatments were used?
9. Did the authors obtain data regarding the known visual quality problems of orthokeratology lenses, such as night vision loss, ghosting, halos, and glares?
10. Correction for inflated alpha should be considered.
Validity of the findings: The authors over-interpreted the results.  First, this study spanned for only 12 months.  Second, most of the parameters were regressing to baseline values, which indicated that the treatment effect of the lenses was decreasing over time.  Third, the authors did not report any side effects and subjective visual quality measurements.  Fourth, the selection criteria and selection process were not clear.
Additional comments: This retrospective study provided potentially useful data for the field.  However, there are some critical issues to be addressed.


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: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is a well written, well executed, easy to read manuscript with no grammatical errors. The Figures and tables were self explanatory.
Experimental design: In this study the authors have made significant effort to study the visual quality of OK lenses worn juvenile patients who were followed up for a year.
The authors need to include the inclusion and exclusion criteria for recruiting the patients for this study.
Validity of the findings: As it has been mentioned in the discussion "the effect of OK lenses on control of myopia progression is well established", hitherto there is no or very little information is available on their effect on visual quality. This makes this study an unique piece of work, however the discussion part still needs to be re-written to highlight the robustness of this work and its significance to make it more interesting for the readers.
Additional comments: Once again I appreciate the authors for their effort taken to address visual quality of OK lenses.


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: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Professional English is used throughout. References, article structure and data mentioned in manuscript are accord with requirements.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: No additional comments


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

Input:
Title: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Writing of the manuscript has been enhanced.  Some important minor points should be address.
Experimental design: As the adverse events in wearing orthokeratology lenses are one of the major concerns, the authors should mention about excluding the cases who experienced any side effects of wearing the lenses. This information is important for the readers to know that the study population was selected partly based on clinical outcomes. Therefore, this should also be discussed as a limitation of this study. The authors also should be advised to acknowledge the potential biases introduced by the selection criteria.
Validity of the findings: The conclusion should be given a timespan of 12 months as shown in this population.
Additional comments: Thanks for the revision.


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

Input:
Title: ONE-YEAR EFFECT OF WEARING ORTHOKERATOLOGY LENSES ON THE VISUAL QUALITY OF JUVENILE MYOPIA: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors have made necessary changes in the manuscript taking Reviewer's comment into account.
Experimental design: I appreciate the author's for including the inclusion and exclusion criteria to make things clear.
Validity of the findings: The discussion part has been re-written in such a way to highlight the significance of the work with substantial citations to the previous work.
Additional comments: I appreciate the authors for making effort to address the Reviewer's comment.


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

Input:
Title: MEASURING URBAN VULNERABILITY TO CLIMATE CHANGE USING AN INTEGRATED APPROACH, ASSESSING CLIMATE RISKS IN BEIJING
Review round: 1
Reviewer: 1

Output:
Basic reporting: good
Experimental design: good
Validity of the findings: good
Additional comments: I am very happy that the authors have addressed my concerns point by point precisely. No further suggestions come from my side. Therefore, I would like to recommend this manuscript to be published.


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: MEASURING URBAN VULNERABILITY TO CLIMATE CHANGE USING AN INTEGRATED APPROACH, ASSESSING CLIMATE RISKS IN BEIJING
Review round: 1
Reviewer: 2

Output:
Basic reporting: I congratulate the authors of the paper as the revised manuscript has improved significantly in terms of quality and clarity.
Experimental design: Method section is now well described and the use of IOM has been clarified.
Validity of the findings: Authors did not aim to validate their findings (same as many vulnerability studies). In fact it is difficult to validate vulnerability as it is an intrinsic quality of a system and not often easily measurable. I was surprised to know that this is the first climate change vulnerability study conducted in an important city like Beijing.
Additional comments: I congratulate you for modifying this paper. Well done!


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: RAMIFICATION HAS LITTLE IMPACT ON SHOOT HYDRAULIC EFFICIENCY IN THE SEXUALLY DIMORPHIC GENUS LEUCADENDRON (PROTEACEAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment; see pdf
Experimental design: No comment; see pdf
Validity of the findings: No comment; see pdf
Additional comments: No comment; see pdf


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

Input:
Title: RAMIFICATION HAS LITTLE IMPACT ON SHOOT HYDRAULIC EFFICIENCY IN THE SEXUALLY DIMORPHIC GENUS LEUCADENDRON (PROTEACEAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: Clear language and generally well-written. Most of the relevant literature has been cited. The manuscript has a professional structure (but see my general comments below).
Experimental design: Very clear experimental design. Well thought out design and implementation. The research question is well defined and mostly well-laid out. Rigorous investigation performed to a high technical standard. Methods well described.
Validity of the findings: The results are conclusive, despite being negative (see my comment below). Conclusion is well stated and links strongly to the original question. I have queried the relevance of one or two figures (see my comments below), but mostly the figures and appropriate.
Additional comments: The study “Hydraulic constraints of reproduction do not explain sexual dimorphism in the genus Leucadendron (Proteaceae)” by Roddy et al. investigates the potential causes of sexual dimorphism in dioecious species of Leucadendron. Specifically, Roddy et al. test the hypothesis that sexual dimorphism in branching patterns is caused by differing hydraulic demand to reproductive structures. The authors pick up on an earlier paper by Harris and Pannell (2010) suggesting that, because female cones require greater hydraulic supply than male cones in Leucadendron, female branches should be less branched to improve “hydraulic efficiency”. To do so Roddy et al. examine the extent of branching in males and females of two Leucadendron species (L. rubrum and L. daphnoides) and compare this to measures of hydraulic conductance. Although the authors find some evidence of an association between hydraulic conductance and branching patterns in L. rubrum and female L. daphnoides (but not males), they conclude that there is no clear relationship between branching and “hydraulic efficiency” in the genus.
I think that the study is well thought out and thorough in its scope. I agree that there appears to be no clear trend between branching (ramification) and hydraulic conductance, and that alternative explanations for dimorphism should be assessed. Even though the results from the study are negative (i.e. the authors find little evidence to support the Harris and Pannel hypothesis), the science is good and the results are interesting.
My biggest issue with the manuscript is that the authors take some time to develop their case, and this makes it hard to follow at times. This extends from the introduction (see my comment below in the specific comments section) to the results/figures (see my comment below). I wonder if the authors might make the manuscript simpler to read/follow by dropping the discussion of trade-offs and/or scaling relationships and focussing more on simpler tests of the hypothesis (e.g. comparisons between males and females)? We only start to see the hypothesis being tested in Figure 5 and Figure 6, and I wonder if the authors can find a way of getting there quicker (see my suggestions below).
Specific comments:
Introduction
Line 78-82: What is the journal policy on citing Honours theses?
Line 90-93: The relevance of the cell size paper to the current study is not entirely clear.
Line 112-116: The relevance of this sentence in the context of the introduction is not entirely clear.
At times the introduction comes across as being quite convoluted, jumping from one aspect of the study to another. For example, the introduction moves from introducing dioecy and its potential impacts on plant hydraulic architecture (very clear, well laid-out), to mentioning trade-offs (and predictions for different levels of analysis), to an attempt to define efficiency, and back to trade-offs. I understand that the authors have tried to be incredibly thorough in their assessment (including developing a better metric of branching), and that this required introducing Corner’s scaling laws to defend their metric etc. However, I recommend that the authors shorten the introduction, sticking more to the point of testing whether there is a relationship between plant architecture and hydraulic conductance.
Results
Line 254-264: This is the real heart of the results section and the authors should consider getting to it earlier (and expanding on it). Do males and females differ in hydraulic conductance?
Generally, I feel that the results section could be shortened or laid out slightly differently. For example, the authors use three figures (Figures 2 through 4) to introduce their trade-off framework, to compare ramification metrics, and to introduce scaling relationships. Some of this information could very well be supplementary material. (e.g. Figure 3 could very well be a supplementary figure. If the authors established a better indication of branching then that presented by Harris and Pannell (2010), then mention this in the methods, and move on to test the interesting ideas.)
Figure 5: Since leaf area is incorporated into the metric of kLA, are these variables not autocorrelated?
Figure 6 is clear. Did the authors consider conducting a simple t-test to establish whether k varied between males and females? Simply showing that k is not related to sex would go some way to establishing whether “hydraulic efficiency” is linked to reproductive investment.
Also, why such a large range in k? Two orders of magnitude seems fairly large. Can the authors comment on this?
Discussion
Nice conclusion.


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: RAMIFICATION HAS LITTLE IMPACT ON SHOOT HYDRAULIC EFFICIENCY IN THE SEXUALLY DIMORPHIC GENUS LEUCADENDRON (PROTEACEAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: meets standards (please see comments to author)
Experimental design: meets standards (please see comments to author)
Validity of the findings: meets standards (please see comments to author)
Additional comments: General comments

The hydraulic consequences of branch ramification, resulting from sexual dimorphism or otherwise, is an interesting and poorly understood topic.  Also, the causes and consequences of sexual dimorphism is also poorly understood in may respects.  As such, I think these data are important and likely to be cited in the future.  The experimental methods appear to be appropriate and well implemented (very nice in fact).  I have two criticisms, which I think can be easily addressed, and are likely not serious flaws providing the authors can indeed address at least the second criticism.

Firstly, I would suggest the authors consider concentrating on the question at hand (e.g., are their important links between specific-conductive capacity and branch ramification), rather than concentrating so exclusively on one particular study, i.e., Harris and Pannell 2000.  Not only does this have the effect of putting Harris and Pannell on “front street”, but I think it detracts from the main results, i.e., it is probably not so important that Harris and Pannell were wrong, but that hydraulic constraints do not explain branching ramification more generally.  I think most readers would agree with this, and could be address quite easily with just a bit of word crafting.

Secondly, and more importantly, it appears as if the x and y axes in Figures 5a, 5d, and 6a are not independent.  In a way, this is not so important for two reasons.  Foremost, in the face of this non-independence, the authors’ hypothesis are still upheld.  For example in Figure 6a, we should expect positive correlation to result from non-independence, but the authors report none, thus supporting their hypothesis that hydraulics do not limit ramification.  Secondly, figures 5a and 5d do not appear to be very important to the hypotheses and it seems to be the differences between the groups (male vs female, and between species) that the authors are interested in, rather than within-group r2 and p values, that are important to the questions being asked.  Nevertheless, unless I’ve misunderstood something (and the axes are indeed independent), reporting r2 and p values from non-independent axes is invalid.

I might suggest to the authors that if they think the relationships reported in these figures are still important to their story, to then use standard major axis regression to address them directly.  For example, if the question in Fig 5a is, “does shoot shoot conductance scale differently with leaf area between male and female plants (or between the two species), then they could plot conductance on the y axis (Q/pressure gradient) and leaf area on the x axis.  Male shoots and female shoots should be plotted individually and the group-specific slopes and intercepts tested using the “sma” function in the “smatr” package.

Specific comments

91: It’s not clear what the independent variables are in this sentence, e.g., is any trait~hydraulic correlation expected to be significant just because there are constraints on cell expansion?  Also, it reads as if the authors expect that these constraints will only influence variation on the x axis (hydraulic efficiency).  This would not result in covariation between x and y nor consistent coefficients between male and female groups if plotted individually.  Perhaps just a bit more elaboration here would help clarify this point.

94-116:  I have a few suggestions just to make the arguments and terms clearer in this paragraph.  Considering that “efficiency” is indeed such a slippery term (whereas conductance has well understood units and meaning), I wonder if it might be better to simply use something like “mass-specific conductance”… or “volume-specific conductance” to give clear and intuitive meaning to the measurement.  Also, I don’t think it is correct to say that conductance divided by “size” (mass?, volume?, height?) is a good measurement of “flux of water per investment cost of moving that water”.  After all, water moves through xylem via a “free energy” gradient (energy is provided by the sun/atmosphere).  Perhaps it might be more correct to simply state that conductance needs to be scaled by a meaningful metric… and it matters what metric we use (i.e., the authors comparison of mass vs size [volume?]).  Or perhaps the authors really mean to normalize k by “the cost of building and maintaining the network”, in which case volume or mass would be more appropriate.

132-133:  In the case of positive correlation, I suppose many “hard tradeoffs” could be erroneously dismissed simply because the magnitude of error often correlates with the magnitude of the measurement.  This would result in predictably different coefficients for the quartiles.  ...just to play Devil’s advocate…

144-145:  There is a mistake in the wording.  If the conductance has been divided by “cross-sectional area” (144), then it will almost certainly scale inversely with cross-sectional area (145).

166: How many days between collection and the day of measurement?

230-231:  I might suggest replacing “it” with “HB ramification” in this sentence, just to be clear.  I can’t really compare Fig 3 with table 1 because male and female plants are not denoted in the column headings of table 1.  Are there p values to report for the male vs female comparisons?

241-241:  I might agree that poor correlation and divergent coefficients among quantiles are indication for an absence of a tradeoff, but I don’t agree that strong correlation and convergent coefficients are indication that correlations are “likley hard tradeoffs”.  I think the authors could correctly say that the evidence is congruent with a hard tradeoff (or similar), but not that x and y are linked via cause and effect.

244-253:  I am a bit confused by the statistics here.  K_LA is K divided by shoot leaf area, right?  And K_CSA is K divided by shoot cross sectional area, right?  If this is not correct, please clarify in the methods.  If this is correct, we should expect negative correlation between these traits simply because the x an y axes are not independent.  For example, in Fig 5a, the exact same variation in shoot leaf area (x) also appears in the denominator of the y axis.  As such, R2 and P values from the within-group correlations are meaningless and should probably not be reported.  HOWEVER, the p values from the across-group comparisons, i.e., comparing first the slope then the intercept (if slopes are similar) using the “sma” function in smatr should still be valid.  Considering that these between group comparisons are the point of the manuscript, I don’t see the non-independent axes as necessarily being a major problem… but I would strongly recommend not reporting r2 or p values from auto-correlated axes, and instead report the differences in slope (and intercept if appropriate) and p values of the between group comparisons.

There appears to be a similar problem in figure 6a.  For example, I think the cross sectional area appears in the denominators of both the x and y axes.  As such, we should expect positive correlation between x and y simply as an artifact of this.  However, the authors would still be correct in pointing out that even though we should expect positive correlation between these traits, they still did not, and thus, there appears to be no strong evidence that more highly ramified branches require greater K_csa… which is consistent with the author’s hypothesis.  Figures 6b, c, and d appear okay though.

299-300:  Again, I think the authors could say that it is not inconsistent with a true tradeoff, but I don’t think this is good evidence that one exists.

Table 1:  It is not clear which values are male and which are female from the column headings.  Also, the table is missing a bottom line.  Would it be possible to report p values for the male-female comparisons?

I would like to thank the authors and editor for giving me the opportunity to review this very interesting study.  I sincerely hope that the authors find at least some of my comments useful.

Sean M. Gleason


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: ACTIVE AND PASSIVE PROCRASTINATION IN TERMS OF TEMPERAMENT AND CHARACTER
Review round: 1
Reviewer: 1

Output:
Basic reporting: First of all, let me thank you for asking me to review this paper.  At the general level it is well written and it addresses most of the literature related to the topic of procrastination. However, the Introduction lacks a consistent logic om how the authors intertwine procrastination-personality-emotional intelligence.
Experimental design: I do think that the research question is well-defined. The authors should develop the limitation section of the paper much further than what is stated in the present version.
Validity of the findings: The findings and conclusions are valid as far as I can see. However, I'm not sure I follow in the rationale about temperament and character profiles. What I can see is that the authors create single scores that then are used to state that, in the case of temperament for example, the higher value means dependable temperament while the lower, the less dependable. I do not see that this is a profile and it does not seem to be used as a profile either. But then again, I might have misunderstood what the authors actually did. In addition, saying that an active procrastinator is creative is a bit problematic...for instance, are all creative individuals active procrastinators? The authors need to adress these type of questions that their findings and design generates.
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: ACTIVE AND PASSIVE PROCRASTINATION IN TERMS OF TEMPERAMENT AND CHARACTER
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English language is at a good level, but it can be improved after the manuscript is improved.
Literature references are satisfactory, with sufficient field background/context provided.
Professional article structure is satisfactory and the article has all the standard sections as well as figures and tables. The data under tables’ and figures’ titles should be transferred to the Results section. Raw data are also shared.
The manuscript includes all results relevant to the hypothesis, but I suggest hypotheses to be better formulated.
Experimental design: This is original primary research within Aims and Scope of the journal.
Research question are not well defined but according to the aim and hypotheses, I assume that they are relevant and meaningful. It is stated how research fills an identified knowledge gap.
The research was performed to a high technical and ethical standard, and methods are described with sufficient detail & information to replicate.
Validity of the findings: My biggest concern is regarding how data were reported in this manuscript and I think there is room for improvement in this part. Once the results are presented in a more understandable and informative manner, the discussion should also be adapted to this. Finally, the conclusions should be better defined and in line with the problems which should be provided.
Additional comments: I have read the manuscript “Active and passive procrastination in terms of temperament and character” several times and I find this to be an interesting research paper that gives a new perspective on procrastination research. I find the research was done properly, but the manuscript in my opinion could be a little better organized and written in the results and the discussion part. Some moderate changes could help this manuscript be more easily understood, more informative regarding the interesting data it presents, and in the end, more easy to read and follow.
As for basic reporting I think that the manuscript is well written, and that the level of English is satisfactory, but there are specific points where some of the sentences could be rephrased (I list them later in specific comments).
As stated above, I think this manuscript has room for improvement, specifically in the relation between aims – results – discussion. It is not quite clear what the aim was because it is described in general, with no specific aims listed. The authors do list the hypotheses, which is good, but the results do not follow those hypotheses clearly and understandably. For some of the analyses it is not clear as to why the authors did them and in what way they help answer their problems, because they do not explain it, and this is not something that will take a lot of time to add. I just think that this should be stated more clearly, because in fact the authors listed 4 hypotheses in the Introduction and then in the Results started answering them in the opposite direction.
You mention 4 hypotheses - I suggest also writing 4 problems first, and then the exact hypotheses – what precisely are you expecting to find regarding those 4 problems, what kind of associations. In the results, after general descriptive part, your result should be presented in a way that makes it clear what you did do answer your problems, one by one, and what the results in this manner show. In the Discussion part you should comment on those results, again in the same order, so it is easy for the reader to follow. An in the end, in Conclusion you should sum up your results as precise answers to your problems stated at the beginning.
All the explanation given under Table or Figure titles should be moved to the Results section.

As for more specific comments, they are listed in a separate file that I will attach.


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: VIRUS-INDUCED GENE SILENCING IN THE PERENNIAL WOODY PAEONIA OSTII
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript reported the application of a TRV-based virus-induced gene silencing (VIGS) in the tree peony species P. ostii. Gene of P. ostii phytoene desaturase (PoPDS) was cloned and then used as a marker to test the TRV-based VIGS system in three-year-old P. ostii seedlings. According to this research, the Agrobacterium-mediated seedling vacuum-infiltration was determined as an appropriate way. After TRV-PoPDS inoculation, a typical photobleaching phenotype was shown in the newly-sprouted leaves of tree peony seedlings. Infection of TRV and silencing of PoPDS were confirmed by RT-PCR and qRT-PCR. In addition, the fluorescent tracer study of TRV-GFP in P. ostii seedlings indicate the potential of utilizing TRV-based VIGS system to analyze gene functions in different tissues.

This manuscript conformed to an acceptable format of ‘standard sections’. Sufficient introduction and background were provided. Figures are relevant to the content of the article, but the description need to be improved. Raw data have been made available. In general, this manuscript is worth of publication, but the English written should be improved and there are a number of ways that this manuscript could be stronger.

Following are some general comments for this manuscript and corrections to English grammars errors and typos that will help improve this manuscript:

Please make sure the usage of spaces between numerical values and unit symbols are correct, as suggested by International System of Units. For instance: “10 min”, “94℃”, “48 h”, “1797 bp” and “2-5 μg”.
Line 25: Replace “significance” by “values”;
Line 26: Replace “current limitation for its” by “limitation for its current”
Line 28: Replace “P. ostii” by “Paeonia ostia”
Line 30: Replace “infection” by “inoculation”
Line 30: Replace “agrobacterial” by “Agrobacterium”
Line 36: Replace “of inoculated plants” by “of plants inoculated”
Line 41: Replace “Virus-induced” by “virus-induced”
Line 42: Replace “Green” by “green”
Line 61: Add “ (P. ostii)” behind “Paeonia ostia”
Line 63: Replace “functional analysis” by “function analysis”
Line 67: Replace the sentence “an efficient genetic transformation system is still not established” by “the lack of efficient genetic transformation system”
Line 77: Please carefully check the description of short interfering RNAs. “21-24 nucleotides” or “21-23 nucleotides”?
Line 75: and Line 369 Please check the publish year, 2003 or 2011?
Line 154: Please double check that it was in a “growth chamber” without shaking.
Line 156: The 600 in “OD600” should be subscript.
Line 165: Delete “The”.
Line 181: Delete “in ”
Line 187: Delete “infected with TRV-GFP, ”
Line 193: Please provide the producer or origin of “anti-GFP antibody conjugated to alkaline phosphatase”, because Bedoya et al did not use alkaline phosphatase conjugated GFP antibody in the referred paper (Bedoya et al., 2012).
Line 209: Replace “and not in” by “but not in”
Line 204: The sentence “These two methods acted on the leaf back and whole plant, respectively.” is ambiguous and confusing. Please revise it.
Lines 205-206: Replace “vacuum way” and “syringe way” by “vacuum infiltration” and “syringe infiltration”
Line 207: Add “Furthermore, ” before “the syringe infiltration”
Lines 213-215: Replace the sentences “Using the transcriptome data obtained from developing leaves of tree peony, we PCR-amplified the open reading frame (ORF) nucleotide sequence of phytoene desaturase (PDS) of P. ostii, annotated as PoPDS, which was commonly used as a reporter for silencing” by “Phytoene desaturase (PDS) is commonly used as a visible reporter for silencing. Based on the transcriptome data obtained from developing leaves of tree peony, we PCR-amplified the open reading frame (ORF) nucleotide sequence of P. ostia PDS, annotated as PoPDS.”
Line 215: Replace “PoPDS encodes” by “PoPDS was predicted to encode”
Line 218: Replace “PoPDS amino acids shared” by “amino acid sequence of PoPDS shared”
Line 220: Replace “peptides of PoPDS” by “amino acid sequence of PoPDS”
Line 227: Replace “seemed” by “seems”
Lines 231-233: It is insufficient here to make a conclusion that “PoPDS of tree peony could be silenced by VIGS and TRV infection was systemically established.”. Authors should draw this conclusion at the end of next paragraph.
Lines 235-237: It is inaccurate to say “DNA fragment of TRV1 and TRV2”. Please totally rewrite the sentence “DNA fragments of TRV1 and TRV2 were detected in TRV empty vector- and TRV-PoPDS-infected leaves, but not in the mock control plants”.
Line 238: “PoPDS insert was detected in the leaves agro-infiltrated with TRV-PoPDS”, but in legend for figure 4 “Semi-quantitative RT-PCR analysis of TRV1 and TRV2 accumulation levels in systemically-infected P. ostii leaves”. Agro-infiltrated leaves or systemically-infected leaves, which one is correct?
Line 243: Please be careful with the use of extreme words in academic writing, such as “definitely”.
Line 249 Add “(dpi)” after “5 days post inoculation”
Lines 262-264: The sentence is difficult to read. please rewrite it.
Line 276: Change “seeding” to “seedling”
Line 280: Change “small-scale” to “limited area”
Line 280: Change “infected with TRV” to “infiltrated with TRV ”, because (as shown in the text) you did not verify the virus infection in plants inoculated via syringe-infiltration method.
Line 295: Replace “Because the reproductive buds of triennial tree peony plants didn’t take shape” by “Because no reproductive buds were formed at this stage”.
Line 296: Replace “in the following work” by “in future work”
Line 301: Replace “possibility of VIGS in tree peony” by “possibility of applying VIGS in tree peony”
Line 303-304: Replace “didn’t except some lesions resembling TRV empty vector-treated leaves” by “showed lesions resembling those of TRV empty vector”
Line 320-321 “viral propagation and the silenced systemic response” “viral propagation and/or systemic silencing response”
Lines 324-325: This sentence is grammatically wrong. Please rewrite it.
Line 325: The reference (Quadrana et al., 2011) sited here is inappropriate. Quadrana et al established a VIGS system based on the transgenic plants expressing exogenous GFP and evaluated its impact on metabolism, they did not mention TRV-GFP.
Line 326: Change “In present work” to “In the present work”
Line 327: Change “abundances” to “accumulation levels”
Lines 331-332: Move the sentence “Previous findings proved that TRV virus possesses the ability to move efficiently within the roots of infected plants (Macfarlane & Popovich, 2000).” To line 329, right ahead “We found a higher…”.
Line 333: Add “for the” behind “underlying mechanism”
Line 335: Please explain the meaning of the phrase “TRV-GFP vector was positive to tree peony plants”
Line 365: Replace “Yanlog” by “Yanlong”

Line 399: Delete the empty line
Lines 402-404, 107, and 328 : Please check the initial and last names of the authors, “Ji T” or “Tian J”? “Shuai Z” or “Zhang S”?
Lines 444-445: Please add the name of book, publisher, and page number.
Lines 468-470 and lines 402-404: reference duplicated.
Line 472: delete one “:”
Line 485” Change “pp.01064.02016” to “853-862.”
Lines 500, 503, and 506: Please add the page numbers

Figure 1d: TRV transcripts accumulation levels in vacuum-infiltrated leaves and syringe-infiltrated leaves are different. It would be better to emphasize this point in the text, since it may provide an reason for the choice of vacuum-infiltration method. “18S-26S-ITS” should be “18S-26S ITS”

Figure 3a: Please describe the meaning of the black box, such as “Black box indicates the PoPDS insert”

Figure 4b: “the primers for TRV2-1 were designed outside the multiple cloning sites (MCS)” should be “the primers for TRV2-2…”.
Actually, since Figure 3 has described the details, there is no need to mention again the primers here. I noticed that the size of TRV2-1 products from TRV-PoPDS-infected plants and TRV-infected plants are different. Would you please give an explanation.

Figure 5: There is no need to mark units to all the scale bars. I prefer to delete all the units in figures and describe them in the figure legend, such as “Scale bars equal to 100 μm (a-f) or 75 μm (g-i)”. “Merge” should be “Merged”.
Figure 5 legend: Replace the whole paragraph by “Confocal microscopy image of P. ostii leaves and roots infected with TRV-GFP at 5 days post-infiltration (dpi). Fluorescence was not observed in leaves (a-c) and roots (g-i) of mock-treated plants.” by “ ”. Replace the sentence “The cell outline (a, d, g, j), the dark fluorescence (b, e, h, k), and the combination photographed in bright field (c, f, i, l) are shown.” by “The bright field (a, d, g, j), the GFP channel (b, e, h, k), and the merged images (c, f, i, l) of the bright field and the GFP channel are shown.”.

Figure 6a: To make the figure easy to read, please delete all the “kDa” behind numbers and left one “kDa” on the top of numbers. Please label the blot with “anti GFP” and the stained gel with “Coom.” or “Loading” at their right side, and then provide explanation of any abbreviated label in the figure legend. I think the line beneath “Mock Leaf Root” should be on the top of “Leaf Root”.
Experimental design: The research is relevant and the experimental design is simple but appropriate.

The following additions would be useful:

Please describe how many replicates are made for each assay.

Lines 181-182: Please make an definition or explanation of “Transient assay of GFP”. In addition, the author only mentioned “leaf cells” here, how about the GFP imaging in root cells? Please describe the details.

Line 231: It would be better to display the images of stable phenotype at 5 months as you indicated in the text, or else please add “(data not shown)”.

In the result section: “Validation of TRV-GFP in P. ostii leaves and roots”. I will recommend to add the TRV infected plants as negative control in addition to mock control, due to the potential nonspecific fluorescence from damaged cells caused by virus infection.
Validity of the findings: Most of the conclusions are well stated and linked to the research performed.

In lines 346-348 of conlusion section, please make sure that the main purpose of the TRV-GFP assay is to reveal “a valid means to a valid means to monitor the viral spread in different tissues”. In my opinion, you try to use this result to prove that TRV is a versatile tool to analyze gene function in different tissues. If so, please rewrite this part.
Additional comments: The authors should carefully correct for all English usage and clarity.


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: VIRUS-INDUCED GENE SILENCING IN THE PERENNIAL WOODY PAEONIA OSTII
Review round: 1
Reviewer: 2

Output:
Basic reporting: Literature related to whether the tree is host for TRV, not provided. Is this a natural host plant?
Much of the data can be moved to supplement and additional data needed for making inferences like 'silencing protocol established in this species'.
Experimental design: Already known methods are used in this study. Authors should have attempted to use different age of seedlings and shown the best stage for the silencing experiment. Statistics and replicate details should have been better described.
Validity of the findings: Extending the VIGS protocol in this tree species is useful. But, authors have simply used already known vector and inoculation protocol and standardized to silence PDS gene. There is no novelty here. Authors should generate more data and validate the functional relevance of an important gene and demonstrate VIGS as tool for advancement of research in this species.
Number of replicates and independent experiments are not clear.
Additional comments: In this manuscript, authors attempted to establish TRV-based VIGS in tree peony. They have taken already available VIGS vector and cloned partial fragment of PoPDS and infiltrated the young plants. They have tried both syringe and vacuum methods for agrobacterium inoculation and state that they found the later works better. They have also traced the GFP signals in roots.

The main concern with this study is the evidences are preliminary and need additional data by showing silencing of other genes involved in specific plant process. Overall, the reproducibility of the silencing data could not be perceived from the evidences presented. How many times each experiment was repeated? It should be stated in the legend.

Line #116, three year or four week old, confusing.
I could not understand how the GFP construct was made and how finding GFP signals in root indicate silencing? Authors’ claims in this regard are vague and evidence is too preliminary.
Fig 5, GFP signals shown are very weak, though the western has detected the GFP protein.
Fig 4, what is TRV2-1 and TRV2-2? I can hardly see photo-bleaching in the leaves.
Authors need to show silencing of genes involved a specific plant process and substantiate the silencing. PDS silencing alone is not adequate to conclude that the VIGS system is established.
Table 1, Figure 1-3 should be moved to supplementary.


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: VIRUS-INDUCED GENE SILENCING IN THE PERENNIAL WOODY PAEONIA OSTII
Review round: 1
Reviewer: 3

Output:
Basic reporting: The lack of effective molecular genetic tools in tree peony limit its genetics, gene function, and molecular mechanism investigations. In the current research, Lihang  Xie et al. have attempted to establish a novel approach by Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) in the tree peony species Paeonia ostii. This is an interesting paper demonstrating TRV-based VIGS technique could be adapted for high-throughput functional characterization of genes in tree peony. In my opinion, it will certainly attract numerous citations in the future. The English in this manuscript is generally correct and intelligible. Also, the scientific background and the research as a whole are sound. Therefore, I suggest accepting this manuscript with some minor revisions.

Please provide detailed information regarding data deposition statement (PoPDS,…) in revised manuscript.

There are still typos and formatting errors in the manuscript. Number and unit should be in between space, for example, “5 min” instead of “5min” in line 133, etc.

Figure 2, sequence IDs should be clearly represented in legend.

Figure 6a, what is the bottom panel? Something like there were no targeted bands in this panel?

The references should be in a consistent format with species name in italics, articles and journal names in similar illustration, etc.
Experimental design: All good
Validity of the findings: All good
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: MATERNAL ANCESTRY ANALYSES OF RED TILAPIA STRAINS BASED ON D-LOOP SEQUENCES OF SEVEN TILAPIA POPULATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors should carefully go through the manuscript again and ensure that due attention is given to the point raised as indicated in the comment boxes attached to the manuscript.


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

Input:
Title: MATERNAL ANCESTRY ANALYSES OF RED TILAPIA STRAINS BASED ON D-LOOP SEQUENCES OF SEVEN TILAPIA POPULATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript title with ‘Maternal ancestry of red tilapias based on D-loop sequences analysis of seven tilapia populations’ written by Jiang et al analyzed the genetic diversity and variants of seven tilapia populations, and further speculated the maternal ancestries of three red tilapia varieties, based on the mtDNA D-loop sequences. The paper finely descripted the experiment design, and also provided a relatively complete genetic profile for the tilapia in China, especially demonstrated the new genetic information for the maternal ancestries of several red tilapia strains. In brief, the research is valuable for understanding the genetic profile of tilapia in China, especial for genetic origins of red tilapias, which could be the support data for the genetic protection and further utilization of tilapia resources in China.  However, there are some mistakes and unclear descriptions presented in this paper.
In my personal opinion, the present paper could be accepted only after minor revision.
The specific advises or suggestions are written as follow:
(1) Line 29: the description of tilapia resource of China, which might not closely with the present study. Which would suggest to replace with ‘many tilapia species or varieties have been widely ….’.
(2) Line 36: the scientific name of red tilapia had been written with mistake, which should be corrected with ‘Oreochromis spp.’.
(3)  Line 63: there are words presented not clear, that is ‘tilapia variants that sport and attractive red coloration.’
(4) Line 80: the three strains are not clear, which are first time presented in the introduction. It is suggested to be rewritten with ‘three strains widely cultured in China (Yang et al., 2016)’.
(5) Line 98: the number of ‘48’ in the head of sentence, should be corrected using the letter form, i.e. forty-eight.
(6) Lines 125 & 128: the written form of software names should be checked carefully, ‘DNAsp5.1’ and ‘Mega 5.05’, should be rewritten with ‘DnaSP 5.1 software’ and ‘MEGA 5.05’.
(7) Lines 128-131: the sentence which suggested to be rewritten with ‘MEGA 5.05 software (--) was utilized to calculate the Kimura 2-parameter (K2P) distances among populations, and to construct the Neighbor-joining (NJ) dendrogram set with 1000 replications of bootstrapping.’
(8) Line134: the software used to construct the haplotypes network, which should be provided in the paper.
(9) Lines 140-142: the description for nucleotide composition of D-loop sequence is incorrect. The D-loop sequence is one kind of double-strand DNA sequence, the composition of A and C should equal to T and G, respectively.
(10) Lines 175-180: the word ‘inter-population’ which suggested to be replaced with ‘among populations’.
(11) Line 184: the subtitle of this paragraph, which suggested corrected with ‘haplotypes network of D-loop sequences in tilapia populations’.
(12) Line 219: the word ‘generation’ should be replaced with ‘generations’.
(13) Line 248 & 251: corrected the genus name and significant probability symbol with italic font, i.e. Channa, and P, respectively.
(14) References related advices, the form of reference cited in manuscript and the references list should be carefully checked according to the journal. such as following:


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: UNILATERAL HAMSTRING FOAM ROLLING DOES NOT IMPAIR STRENGTH BUT THE RATE OF FORCE DEVELOPMENT OF THE CONTRALATERAL MUSCLE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The aim of the study was to compare the effect ten × 30-s SAFR vs 10 min rest on the contralateral (non-intervened) muscle’s RFD, isometric MVC and corresponding EMG. The results demonstrated a significant improvement in the hip flexion ROM and a decrease in F50 following SAFR, without any change in MVC force and EMG. The manuscript is well written and the experimental protocol is pretty clear. However, a major limitation with the study is that although simple measurement techniques (e.g. MVC force, RFD, EMG, ROM) were used, the authors have provided a lot of speculations regarding the underlying mechanisms (mainly neural factors) contributing to the results. Few of these issues have been highlighted below.
General comments
1)	Introduction line 103-105: Based on what “neural mechanism” do the authors believe that SARF could alter muscle performance in the contralateral limb? This question was not elaborate in the introduction (and even in the discussion). In lines 103-105 as well as 125, the authors have provided very vague speculations about the “neural mechanisms” and ”rolling induced neural inhibition” however the reviewer could not find any evidence for this across the entire manuscript, particularly because none of the measurement techniques used in the study would directly (or even indirectly) assess these potential mechanisms.
2)	Introduction line 121-125. It is not clear why the authors needed to “reexamine” the effect of SAFR on flexibility and MVC force of knee flexors. In the previous study you had 23 participants and in this experiment 34. Did you add 11 participants to your data pool, or these are 34 new participants? In either of the case, please elaborate why the reexamining the ROM and MVC force was necessary?
3)	Introduction line 133-135. How could your data provide a better understanding about “potential neural inhibition …. and recovery of the immobilized limb”?
4)	Methods, line 187. The examiner who measures the ROM should have been blinded to the intervention otherwise this could have affected the outcome measures.
5)	Discussion, line 295-298. Although the authors found only a time effect, they decomposed the pre vs. posttest values in each intervention session and then compared the magnitude of effect sizes between the two conditions. Theoretically, the authors are NOT allowed to decompose the time effect into two separate groups. In order to do this, you need to have an interaction effect. In addition, please consider that d = 0.34 and 0.24 are already showing very small effects, so it does not make sense to compared those with 0.09 and 0.11 (while statistically there is no difference between them!).
Experimental design: Specific comments:
Abstract: The conclusion is over-reaching (see my general comment #1)
Introduction, line 108. The word “unilateral” should be replaced with “contralateral”…
Introduction, line 108-109. While Cavanaugh et al. (2017) did not find any effect on F200, it is not clear how the authors expected that F50 or F75 would result differently…
Introduction, line 112-115. This sentence needs to be revised. The structure of the sentence looks weird.
Introduction line 118: How much is the ecological validity of 10 sets × 30 s FR? This is a limitation to the results.
Methods, line 162. Is there any reason why the rolling and testing legs were not selected randomly between participants?
Methods, line 169. The order of ROM and MVC force after each intervention indicates that the MVC force of the tested leg was performed with ~ 45 s to 1 min delay, whereas in 296 you use the word “immediately” which is not correct.
Methods, line 187. The examiner should have been blinded to the protocol, otherwise this should be mentioned in the limitation.
Methods, line 189. What does slight stretch means? How much was the reproducibility of the ROM, RFD and MVC data? The ICC, CV and standard error of measurement should be reported.
Methods, line 196. Based on what criteria this 2 degrees was selected? Please provide a reference.
Methods, line 207. You used 3 contractions at 50% of MVC for warm up. What is your reference for this protocol?
Statistical analysis. Did you test the assumption of normality and sphericity for all dependent variables? Also, for the time and condition effects, you should use Bonferroni, as paired t-test is only applicable with an interaction effect.
Results line 251. There is only one figure in the manuscript (not figures)
Results line 257. It is a bit strange that the P < 0.001 but the d is very small!
Results. Overall, regardless of the p values, the effect sizes for all outcome measures (even significant ones) are very small which questions the strength of the intervention compared to the control condition.
Discussion, line 286. The increase in ROM following rolling massage is a very controversial concept, the word “as expected” indicates as if the authors have accepted the positive effect of SARF without any hesitation …
Discussion, line 295-302. Based on statistical analysis guidelines, the entire paragraph should be deleted (see my general comment # 5).
Discussion, line 309. Again, what neural factor?
Discussion, line 314. what afferent receptors? Very speculative though!
Discussion, line 316. What somatosensory training?
Discussion, line 321. Regardless of the provided explanation, that is very over-reaching, the references # 33 and 34 are both measuring the effect of static stretching, but you are using the evidence to infer the influence of SAFR. The two technique could have totally different effect.
Discussion, line 348. The stretch tolerance was not defined well, although as far as the reviewer is concerned, this is the main reason for the observed results rather than inhibitory neural effect.
Figure 1 does not elaborate anything (because it shows the pooled data). I would delete it.
Validity of the findings: The ICC, CV and standard error of measurement for RFD, MVC force and ROM should be reported.
Additional comments: No additional comments


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

Input:
Title: UNILATERAL HAMSTRING FOAM ROLLING DOES NOT IMPAIR STRENGTH BUT THE RATE OF FORCE DEVELOPMENT OF THE CONTRALATERAL MUSCLE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors present a manuscript on the effects of a hamstring foam rolling intervention on the range of motion, strength, and rate of force development of the contralateral limb. In general, the manuscript is clear and well-written. However, I have some suggestions that I believe will improve the quality of the paper and make some parts more transparent.

General recommendation
- When discussing statistical significance, I recommend avoiding the term "significant" in isolation (throughout), and to precede it with "statistical". This will ensure clarity in that you are talking about statistical, not clinical, significance.
- I suggest avoiding the term "muscle activity" and "activation" when referring to EMG results (see https://www.frontiersin.org/articles/10.3389/fphys.2017.00985/full)
- All discussion and reporting should be relative to the control group. I recommend not reporting inferential statistics for time or group main effects. This ignores the purpose of a control group. See comments for references.
Experimental design: No comment.
Validity of the findings: The analyses can better address the research question/design by focusing on the effects relative to the control. Discussion of the results can be more specific.
Additional comments: FIGURES AND TABLES
Figure 1 - Rather than bar plots, I recommend a plot that allows the reader to see the individual data. For example, a Gardner-Altman plot, a Cumming plot, or a slope graph. Estimationstats.com may be a good resource for inspiration.

ABSTRACT
- The abstract is subject to change given some of my other comments. I will reserve comments until all analyses are clearly relative to the control group.

INTRODUCTION
Paragraph 1 - At the authors' discretion, I suggest avoiding terms that suggest foam rolling's mechanism is tissue/mechanically-mediated (e.g., "myofascial release", "soft tissue mobilization", etc.). For instance, referring to foam rolling as a method of "massage" would be just as effective and more cautious.
L77 - I recommend avoiding anecdotal evidence. The increasing number of studies on the topic is likely enough to suggest that it's of interest for researchers and clinicians.
L88 - Not all studies have found that foam rolling does not impair performance; for example, Monteiro's work. As such, this can be reworded more cautiously, "without necessarily impairing…"
L91 - I think "heterologous" is more appropriate than "heterogeneous"
L99 - I recommend rewording this sentence to suggest that it is not the nociception in and of itself that leads to improved stretch tolerance, but potential downstream effects of nociception (e.g., descending inhibition)
L100 - "mechanical sensitivity" or "stretch tolerance" may be more precise than "pain sensitivity"
L102 - Is it the pain or discomfort that is modulating excitability or foam rolling (from a purely methods standpoint)? It is not clear to me that pain/discomfort—effects of the intervention—are causing the excitability changes. As such, I think it's more accurate to attribute these effects to foam rolling (the intervention).
L109 - I recommend explaining what is meant by "neural factors"; e.g., MU recruitment or rate coding?
L112 - More sensitive parameter than peak force for what, exactly? Altered excitation affecting the force generating ability of nonlocal muscles? Please clarify.
L115–117 - Do they always serve as better predictors? If not, it may be best to modify this sentence to read "often serve…"
L122 - "thus" is not needed
L124 - What about the semimembranosus?
L128 - Different or statistically different?
L129 - I don't think filling a gap is in and of itself important. Importance may be improving our basic knowledge of something or having clinical/real-world relevance. The following sentences start to get at this. As such, I would reword this sentence slightly. For example, "This will work build upon previous work by explicating the effects of high-volume, …"

METHODS
L139 - Why 34 subjects and the uneven split between men and women? Was it out of convenience? Were participants familiar with foam rolling?
L161–163 - Why was the intervention applied to the dominant rather than non-dominant thigh as opposed to, say, randomizing?
- Why was such a high volume of foam rolling of interest?
- For all measurements, were the observers/investigator(s) blinded at all?
- Do you have test-retest reliability information?
L196–197 - Why use the average rather than modeling the variance (e.g., using a mixed-effects model)?
L196 - How many extra trials? Why two degrees?
L223 - Do you happen to have impedance values?
L226 - 20000 Hz or 2000 Hz? The former seems high.
L229 - What type and order were the filters?
L239 - Why the linear portion from t = 0 to X rather than the dF/dt evaluated at t = X?
L246 - Are paired t-tests really of interest over the ANOVA interaction? It seems that these ignore the control group, so the results wouldn't seem very interesting.
L247 - Did you control for multiple comparisons?
L249 - How was Cohen's d calculated?
L249 - Cohen's interpretations were not intended for exercise science. Hopkins' interpretations may be preferred?  https://www.sportsci.org/resource/stats/effectmag.html

RESULTS
- For all outcomes, please present the F statistic, its degrees of freedom, and the p-value.
- I suggest focusing on the ANOVA interaction (or difference between deltas) rather than main effects and post hoc testing. Inferential statistics within a condition ignore the purpose of the control group. See:
Bland, J. M., & Altman, D. G. (2011). Comparisons against baseline within randomised groups are often used and can be highly misleading. Trials, 12. doi:10.1186/1745-6215-12-264.
Bland, J. M., & Altman, D. G. (2015). Best (but oft forgotten) practices: Testing for treatment effects in randomized trials by separate analyses of changes from baseline in each group is a misleading approach. The American Journal of Clinical Nutrition, 102(5), 991–994. doi:10.3945/ajcn.115.119768.

DISCUSSION
L288 - Because joint angles are not ratio scale, you cannot have a percent change. Please remove the "7%"
L295 - This should rely on the interaction effect. If you wish to use an estimation approach, I recommend re-analyzing the data to get the estimates and their uncertainties you are interested in.
L304 - Trends do not exist within the NHST framework. As per my previous comment, if you wish to use an estimation approach, I recommend re-analyzing the data
L305–309 - Are these relative to the control group or independent of the control group?
L313–324 - I think the explanation here can be made more specific. There are different terms, lines of evidence, and hypotheses being presented, but many of them are not pointed and do not seem contiguous. For instance, there are many afferent receptors—to which ones are you referring? Reference 35 deals with reciprocal inhibition; how does this relate? I think this discussion can be made tighter.
L347–350 - Please be more specific than "neural inhibition to the central nervous system" – by what mechanism and how would this occur? It's okay to speculate, as specificity in the discussion will make it more interesting and insightful.


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: UNILATERAL HAMSTRING FOAM ROLLING DOES NOT IMPAIR STRENGTH BUT THE RATE OF FORCE DEVELOPMENT OF THE CONTRALATERAL MUSCLE
Review round: 2
Reviewer: 1

Output:
Basic reporting: Thank you for making the revisions.
Experimental design: No further comment
Validity of the findings: No further comment
Additional comments: There is one more comment regarding the sentence stated in line 290-293. This should be the other way round. When ANOVA shows significant main effects of times or conditions, Bonferroni post hoc comparisons (appear below ANOVA table) are reported. When an interaction effect is observed, paired t-tests with Bonferroni corrections are applied.


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: UNILATERAL HAMSTRING FOAM ROLLING DOES NOT IMPAIR STRENGTH BUT THE RATE OF FORCE DEVELOPMENT OF THE CONTRALATERAL MUSCLE
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I would like to thank the authors for addressing my comments. I feel that the manuscript is close, but I have a few more comments and clarifications that I feel can improve the manuscript.

INTRODUCTION
- These are small comments that I think can make the introduction more focused:
L106–109: This sentence seems opaquely speculative, and I am not sure that it adds much to the paragraph.
L110–116: These sentences are rather specific for what we know about foam rolling. It may be best to remove them and stick with the evidence that we have. E.g., "Recent work suggests that foam rolling can have nonlocal effects…"
L131–134: I think this is worth rewording or restructuring. For instance, EMG amplitude is a measure, not a mechanism, and higher level/more central mechanisms may be the mechanism by which the downstream effects (e.g., MU recruitment) occur.

METHODS
L254: I checked Delsys' website, and they do not have 20 kHz listed (https://www.delsys.com/bagnoli/). Please triple check this! If you're 100% sure, you can ignore me; I just have not seen sEMG sampled at that rate before.
L277: Please double check this equation. Shouldn't it be SD*sqrt(1-ICC)?
L280: The model (fit) residuals must be normal, not the DVs.
L290: Why 0.63 and 1.15? Shouldn't it be 0.6 and 1.2?

RESULTS
- Please include both numerator and denominator degrees of freedom for the F-tests
- I encourage the authors to remove the inferential statistics for the within-condition outcomes. This is ultimately the authors' and editor's decision, but I will reiterate that prominent biostatisticians have been discouraging within-group (i.e., time or condition) comparisons such as these. The focus should be just on the interaction effect, as this is what addresses the research question. Descriptives are appropriate for everything else:
Bland, J. M., & Altman, D. G. (2011). Comparisons against baseline within randomised groups are often used and can be highly misleading. Trials, 12. doi:10.1186/1745-6215-12-264.
Bland, J. M., & Altman, D. G. (2015). Best (but oft forgotten) practices: Testing for treatment effects in randomized trials by separate analyses of changes from baseline in each group is a misleading approach. The American Journal of Clinical Nutrition, 102(5), 991–994. doi:10.3945/ajcn.115.119768.
- Can you include the point estimate and standard error for the interaction?
- The plot looks great, but it should be for the interaction effect (difference between the deltas). The within-condition effects are not of interest.

Discussion
L346: Were not statistically significantly altered (lack of stat significance is not evidence for null; see https://www.nature.com/articles/d41586-019-00857-9)
L351: Where are you getting the Cohen's d from? Is this adjusted for the control condition?
L352–353: This sentence reads awkwardly. Please revise slightly
L354: If you update the estimation plots for the difference of differences, this sentence may change slightly. That said, I like that you're looking at your data to infer effects rather than just statistical outcomes (e.g., p-values).
L360: Are these corrected for the control condition?
L384–387: The Nijs paper deals more with chronic nociception. Consider either the reviews by Bialosky or the review by Vigotsky & Bruhns (2015) on manual therapy mechanisms. The latter may be more relevant for neurophysiological mechanisms and DNIC. The sentence will likely change slightly to accomodate the information in the cited review (e.g., the effects being in S1 vs. elsewhere).
L409: It seems uncertain if the effects are occurring in the cortices or elsewhere. It might be best to be a little bit more vague here; e.g., "have induced greater inhibition"

Conclusion
- I do not think discussion of possible mechanisms is necessary here. I would stick to what you found and can conclude from your data. You can suggest areas for future work, but I suggest avoiding heavy speculation.


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: UNILATERAL HAMSTRING FOAM ROLLING DOES NOT IMPAIR STRENGTH BUT THE RATE OF FORCE DEVELOPMENT OF THE CONTRALATERAL MUSCLE
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I would like to thank the authors for their patience and thoroughly addressing all of my 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: PERFORMANCE EVALUATION OF DEEP NEURAL ENSEMBLES TOWARD MALARIA PARASITE DETECTION IN THIN-BLOOD SMEAR IMAGES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Clear and unambiguous, professional English used throughout.
Literature references, sufficient field background/context provided although more recent literature (2018)  should be included
Figures and tables are appropriate and complement the text. More self-descriptive caption should be included.
Self-contained with relevant results to hypotheses
Experimental design: Original primary research within Aims and Scope of the journal.
The article clearly states the contributions of the article however the research question is not so clear, is this a review of DL ,methodologies or are you presenting a new method?

Rigorous investigation performed to a high technical & ethical standard including a deep analysis of the outcomes.

Methods described with sufficient detail & information to replicate.
Validity of the findings: Novel comparison of methodologies and novel framework with lot of details for a replication. Also a web-based model is proposed.
Data is available on-line from a previous work and the statistical analysis is well done.
Conclusions and contributions are well stated
Additional comments: The paper is really interesting and a great analysis of the state of the art on the use of DL for malaria detection. However I missed more recent reference since most of them are from early's 2000.
The captions from the figures and tables are  in general poor, it would be easier to follow if they were self-explanatory.
Line 85 proposes protocols from WHO 2010, why not WHO 2018?
Can you give some references for this sentence: "2015). In particular, convolutional neural networks (CNN), a class of DL models have demonstrated promising results in image classification, recognition, and localization tasks"?
Between lines 125 and 140 you compare CNN with other methods and give the accuracy of the CNN approach, what is the performance of the other methods such as SVM? It is not clear the improvement achieved.
Lines 142-143 presents your previous article, could you make clear what is the novelty in this work?
In the Material and Methods section I am missing a more general overview of the whole process since sometimes is difficult to follow.. A flowchart would help. Indeed I am not sure if figure 2 is presenting this but the steps in figure 2 do not correspond with subsections. In fact Figure 2 is not well explained in the text
Line 232 could be a break line.
Line 283 states that you used both custom and pretrained models, this is not clear until this moment. Giving and explanation of the whole process at the beginning of the section will also clarify this.
At the beginning of the results section you could also states what you want to analyse
Tables 4 and 5 can be fuse in one table.
Line 435, what are you losing?
Rewrite sentence in line 436.
Figure 11 and table 6 can be fuse.
I don't understand what you are trying to show in table 7
Which model are you using in the web-based model?
Where can we find the web-based model?
Table 8 should be include in the results but not in the conclusions.
Clarify sentence in line516


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: PERFORMANCE EVALUATION OF DEEP NEURAL ENSEMBLES TOWARD MALARIA PARASITE DETECTION IN THIN-BLOOD SMEAR IMAGES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Please check overall summary (below).
Experimental design: Please check overall summary (below).
Validity of the findings: Please check overall summary (below).
Additional comments: The authors have evaluated the performance of model ensembles in an attempt to reduce variance and optimally combine the models’ predictions toward classifying parasitized and uninflected cells in thin-blood smear images. The study demonstrates significant improvement in performance, compared to the state of the art and would appeal to the research community. However the authors have to clarify on a few aspects pertaining to the methods, results, and related discussions.

Specific comments:
1. How did the authors detect and segment the red blood cells? How did they evaluate the segmentation accuracy?
2. How did the performance of the proposed ensemble compare to that of the human readers?
3. How effectively could the proposed ensemble be deployed into mobile devices or cloud for the ease of predictions?
4. How did the authors settle for the optimal t-sne hyper parameters for the current study? Could the process be generalized?


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: PERFORMANCE EVALUATION OF DEEP NEURAL ENSEMBLES TOWARD MALARIA PARASITE DETECTION IN THIN-BLOOD SMEAR IMAGES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors addressed all the comments. The paper is easy to read, very interesting and presenting a model with great results. Congratulations!


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: PERFORMANCE EVALUATION OF DEEP NEURAL ENSEMBLES TOWARD MALARIA PARASITE DETECTION IN THIN-BLOOD SMEAR IMAGES
Review round: 2
Reviewer: 2

Output:
Basic reporting: Good paper.
Experimental design: Accept
Validity of the findings: Accept
Additional comments: The paper has been revised well.