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: METABOLOMICS WINDOW INTO THE ROLE OF ACUTE KIDNEY INJURY AFTER CORONARY ARTERY BYPASS GRAFTING IN DIABETIC NEPHROPATHY PROGRESSION
Review round: 1
Reviewer: 1

Output:
Basic reporting: •	There are some typing errors, so I would appreciate if the authors could check the entire manuscript through the text. Here are some examples:
-line 35: in the Abstract, please specify Diabetes Mellitus before using DM
-line 41: in the Abstract, “paitents” instead of “patients”
-line 71: “for perdicting development of…” instead of “for predicting development of…”
-line 90: “progrocess” instead of “progress”
-line 199: “metabolicpathways” instead of “metabolic pathways”
-line 321: “Key TCA cycle metabolites includes…” instead of “Key TCA cycle metabolites include…”
-line 327: “AA-tyrosine was decrease….” instead of “AA-tyrosine was decreased….”
-line 331: “reported in associations with….” instead of “reported in association with….”
-line 352: “one isomeric forms….” instead of “one isomeric form….”
……….

•	As regard the Introduction, I think the authors should enrich that section, since there are interesting manuscripts to refer to and to comment on. As an example, I may suggest to mention and to discuss on the evidences highlighted in the manuscript by Wang TJ et. al., (Metabolite profiles and the risk of developing diabetes. Nat Med (2011)), by Zhang J et. al., (Metabonomics research of diabetic nephropathy and type 2 diabetes mellitus based on UPLCoaTOF-MS system. Anal Chim Acta (2009)), as well as the findings of “Proteomic and metabolomic characterization of diabetic nephropathy” (Acta Diabetologica (2018)) by Rossi C et.al.. I am sure comments on already described metabolomics characterization of DM and DN would better demonstrate the potential of metabolomics strategy by mass spectrometry in the study of such disorders.

•	Figure 3 is difficult to appreciate in term of resolution, in particular Panel d and e, referring to the KEGG metabolic pathway and statistics of KEGG enrichment are both impossible to read.

•	Figure 4, the colour code of each Panel are confusing: it seems that blue means A group and orange B group, but in the other Panels blue and orange code are used for up- and down-, and for AKI and NAKI. Thus, a reorganization of Figure 4 is strongly recommended.

•	Also Figure 5 is not clear. Firstly, it is difficult to understand that the first 5 scatter plot in panel c are referred to Venn diagram in Panel a, and the last 4  scatter plot in panel c are referred to Venn diagram in Panel b. Moreover, significance information of t-test is missing in the comparison. It is also not clear why showing the third comparison (the green one) in the Venn diagram of panel a and b. Thus, I suggest a better explanation of the data processing.
Experimental design: •	In the Materials and Methods section, in particular in “Targeted metabolomics analysis of the validation method”: it is really difficult for the reader to understand that this is a quantitative analysis.  No analytical parameter, such as linearity, concentration range of interest, precision of the method, is described and no reference to a previously validated method is reported. This is a really important point.
Validity of the findings: Results, as shown in Figure 5 are not clear. Firstly, it is difficult to understand that the first 5 scatter plot in panel c are referred to Venn diagram in Panel a, and the last 4  scatter plot in panel c are referred to Venn diagram in Panel b. Moreover, significance information of t-test is missing in the comparison. It is also not clear why showing the third comparison (the green one) in the Venn diagram of panel a and b. Thus, I suggest a better explanation of the data processing in Figure and through the text.
Additional comments: I am writing about the manuscript of Jiayi Wang et. al. entitled “Metabolomics window into the role of postoperative acute kidney injury in chronic kidney injury among diabetes mellitus patients”.

Through an untargeted metabolomics strategy, metabolites statistically distinguishable were highlighted in postoperative urine samples compared to preoperative urine samples. Following, to better understand the role of identified AKI markers in chronic development of renal injury among T2DM patients, the over-highlighted metabolites were quantified by a targeted metabolomics approach.

Overall, this research article is well written and the aim of the manuscript is well described, bringing relevant observations on metabolomics approach as a valuable tool in the discovery of novel biomarkers of Diabetes Mellitus and Diabetic Nephropathy. Anyway, I have some comments and some aspects to be clarified that would be taken in consideration to improve 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: METABOLOMICS WINDOW INTO THE ROLE OF ACUTE KIDNEY INJURY AFTER CORONARY ARTERY BYPASS GRAFTING IN DIABETIC NEPHROPATHY PROGRESSION
Review round: 1
Reviewer: 2

Output:
Basic reporting: Well-structured and designed manuscript/study, but following issues should be addressed.

#Title should include CABG, not just postoperative AKI.
Experimental design: #Why eGFR was calculated using MDRD, not CKD-EPI?
- over 60 ml/min/1.73m^2, CKD-EPI is accurate and recommended.

#Collection of urine: is there no emergent surgery? (preoperative samples in emergent CABG could not be used

#24h postoperative urine collection was done from immediate postoperative to 24 hours. Is this right? When the urine collection was started? Please describe the method in detail.

#How the author controlled the severity of AKI (e.g., staging of AKI)?
The severity of AKI could affect the relationships and findings.
Validity of the findings: #Please describe following information
- Information on CABG (IABP, CPB, surgery time, and so on)
- Information on diabetes (% of diabetic retinopathy)
- Information on others (anemia, albumin, Hx of ischemic heart disease and stroke)

#How was the recovery of AKI? The levels of metabolites were associated with recovery of AKI or progression to CKD (e.g., 3 month-Cr level)?

#Figures: P values should be described in figures (esp. Figure 5).
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: METABOLOMICS WINDOW INTO THE ROLE OF ACUTE KIDNEY INJURY AFTER CORONARY ARTERY BYPASS GRAFTING IN DIABETIC NEPHROPATHY PROGRESSION
Review round: 2
Reviewer: 1

Output:
Basic reporting: #Title:
Metabolomics window into the role of acute kidney injury after coronary artery bypass grafting in chronic kidney injury among diabetes mellitus patients

Authors included diabetic CKD patients? The mean eGFR was 93 ml/min/1.73^2. All of them had persistent proteinuria? Authors mentioned they included T2DM patients with normoalbuminuria.

Title should be corrected.
Experimental design: They used AKIN criteria for defining AKI, but the KDIGO guideline is a recent one. Please use this criteria.
Validity of the findings: No comment
Additional comments: Most of issues are well 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: METABOLOMICS WINDOW INTO THE ROLE OF ACUTE KIDNEY INJURY AFTER CORONARY ARTERY BYPASS GRAFTING IN DIABETIC NEPHROPATHY PROGRESSION
Review round: 3
Reviewer: 1

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


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND CHARACTERIZATION OF TCP FAMILY GENES IN BRASSICA JUNCEA VAR. TUMIDA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English language is clumsy and should be improved.
The literature is poorly explored. Several references are lacking, along with a better Discussion.
Figures and tables are in general ok (although I detail some concerns for the authors)
Experimental design: Ther study is relevant for the Journal scope. For this reason, a second chance should be given to the authors.
I have some methodological concerns that can be addressed by the authors.
Validity of the findings: I suggest that the findings should be further explored and analysed, specially miR319 targets within the TCP family.
Additional comments: The present study by He et al., reports a genome-wide characterization of TCP transcription factors in Brassica juncea var. Tumida, combining phylogenetics and gene expression data.
The approach of the manuscript may provide new valuable insights in line with the scope of the journal, and also may set the basis for future investigation of the function of TCP members in B. juncea. Nevertheless, I consider that the manuscript can not be accepted in its current version, but a new improved manuscript can be considered for publication in PeerJ.
My main concern is the phylogenetic analysis performed. The authors performed a neighbor-joining tree, which is a distance method, not a cladistic analysis. This is a common error within the comunity of plant molecular biologist. I strongly suggest that you perform a phylogenetic analysis through Maximum likelihood, Bayesian, or even Parsimony. This can change the topology of the tree presented in Fig 2. In fact, authors can use the ML o Parsimony search implemented in MEGA, which was the software of your choice. Lack of experience in phylogenetics is reflected by the fact that (according to the legend of Fig 2A) you presented the boostrap tree, which only serve to obtain clade support, but not to be used as the final topology of a phylogenetic analysis. This is a conceptual mistake. Given that the gene cluster obtained may change after you perform a phylogenetic analysis, the outcome and conclusions may also suffer modifications.
There also other relevant points that need consideration:
1- Literature used is not extensive as it should be, taking into account the knowledge about TCP nowadays.
2- The English language is clumsy and should be highly improved to ensure that an international audience can clearly understand your ms. Although I am not an English native speaker person, some parts of the text were extremely difficult to reach your point. I made the effort of suggest some corrections (see minor comments & corrections), but I consider this is a major pitfall of the ms.
3- Authors claim that in B. juncea there are no homologous genes of AtTCP3, 4, 10, which are direct targets of miR319. These should be further explored given the possibility that TCP family in this species seems to scape the post-transcriptional control by miR319. Authors should confirm this result and discuss it further. In case you find some BjTCP target candidates for miR319, you should at least present an alignement in a figure.
Minor comments & corrections:
Title: replace “characters” with “characterization”.
Introduction:
P6, line 35
…that is reported to BE involved
Also, wrong use of the singular
P6, line 41
Missing space. The same mistake is everywhere in the text.
loop(
P6, line 44
The core sequences described are essentially the same (GGNCCC). Also, class I site is incorrect according to recent reports of TCP, revise with updated literature.
P6, line 48
Misspelling; controlingl
P6, line 51
Incorrect interpretation of Niwa et al. 2013. Although they proved FT and BRC1 interaction, they do not evaluate the importance of this interaction in floral development. Rephrase
P6, line 52
binding with. Incorrect preposition
P7, line 65
Remove the s after sativa
P7, line 87
Awkward use of English.
Suggestion for improvement:
SINCE As the whole genome of tumorous stem mustard was sequenced (Yang et al. 2016), a genome wide analysis of TCP genes was PERFORMED FOR the first time to perform in this research.
P8, line 92
The results can provide valuable information for the classification of BjTCPs and lay the foundation FOR of exploring the mechanism of stem swelling and flowering TIMEregulation regulated ORCHESTATED by TCP proteins in tumorous stem mustard.
Material and Methods:
P8 line 98.
What do you mean with constant light? A 16/8 h light/dark photoperiod is not constant light!. You may refer to temperature instead.
P8 line 103.
No N2 snap frozen first?
P8 line 112.
Use plural in sequenceS
Use past tense for donwload
P8 line 123.
…a maximum number of 12 motifs WAS SET.
P8 line124
Reference for ProComp9.0??
P8 line126
Use cursive for “cis-acting”
P8 line131
Reference for MapInspect?
P8 line133
…WAS can downloaded…
P8 line134
accession number were SRX108496
Results
P10 line 167
…were ARE transcription.
The generalized conclusion of line 166-168 must be rephrased, considering that the authors found some BjTCPs outside the nuclei. In addition, what do you mean in Table 1 with “none” in subcellular localization for BjTCP13a-c?. You didn´t found the protein inside the cell at all? This point should be further explained. I am not familiarized with ProComp9.0, but it is expected that the protein is present somewhere in the cell. This data comes from empirical evidence or is just an in silico prediction?.
P11 Line 179
The absence of homologous sequences of AtTCP2-4 and 10 may be interesting to explore further considering that these proteins are targets of miR319. Other genome wide analyses reported of TCP proteins usually check the conservation of miR319 targets (even in plant species outside the Brassicaceae family). I strongly suggest you do the same. In case you confirm that there are no miR319 targets in BjTCP, these would become an interesting outcome (that should be explored and disscused). Also, under this scenario, it may be relevant also to check the existence of miR319 in Bj.
P11 Line 183-185
The grammar in this phrase is clumsy. Also, you must avoid the use of constrictions like “couldn’t”.
P12 line 208
…and BjTCP18a-d were HAVE more…
P12 line 209
“Chinses”???
P12 line 209
Use italics for BrTCP1a,b (and everywhere in the text)
P12 line 209
exon number form FROM other
P12 line 208-211
The grammar of this phrase should be revised.
P12 line 212
their homologous genes IN B. rapa BrTCP18a and b
Figure 3: Indicate which motif/s include the TCP domain. Remove the Green box with the legend CDS in 3C.
Also, did BjTCP21b have the TCP domain???
P12 line 231
was WERE also present
P13 line 237
were also be found
line 242
on A the previous
line 245
“DA simple” means DY samples?
Line 255
The criterion used to select genes for qPCR is correct, if the sequence similarity is at the promoter regions particularly. This should be explicit.
Line 258-259
Missing Word before “expected”?? AS??
DISCUSSION
Line 273
..some genes were duplicated are doubled in the..
Line 277 ..identified. We…
Line 282-284
It is difficualt to interpret what is the author’s point, given grammar errors.
Line 287 (suggestion for improvement)
…we guess SPECULATE THAT these similarities WITHIN cluster OF homologous genes members SUGGEST THAT THEY might play similar function during tumorous stem mustard growth and development.
Line 291
No need to cite Figures in Discussion section, unless this is indicated in the instruction for authors of the Journal.
Line 294
Given the high copy number through duplication of TCP15 in Bj, along with the expression data analysed, it would be interesting to discuss with the reported functions of AtTCP15.
Suggestion for improvement
AS MENTIONED ABOVE, Tthere are 24 TCP genes in Arabidopsis. Some corresponding homologous OF TCP genes WERE cannot fOUnd in tumorous stem mustard, such as TCP2-4, 6, 11 and 16, PROBABLY DUE TO GENE LOSTwhich may be caused by the losing eventS during the evolutionARY process.
Lines 299-303
This claim is incorrect: “In theory, each of the Arabidopsis TCP genes has two homologous genes in the stem mustard...”
Why Bj has to have the double of genes than Arabidopsis?? Take into account thay the lineage of the family where both species belongs split long ago. Remove or rephrase!
Line 303
“doubling”? replace with duplication.
Also, a connector is missing after “,”
Line 313
were demonstrated REPORTED for
Suggestion for improvement: BRC1/TB1 orthologS with TB1 and rice TB1 that may play a similar role in development of the primary shoot architecture
Line 320
But There are four BjTCP18s homologous genes in tumorous stem mustard, which may HAVE BEEN formED by gene duplication.
Functional differentiation may occur between the four TCP18 genes. From GIVEN their DIFFERENTIAL expression patterns, these four genes are not consistent during tissue development.
LINE328
INDICATEd
Line 329
Suggestion for improvement (if I get your idea)
According to reports, there are 16 varieties of mustard species identified AND USED FOR FOOD CONPSUMPTION, in which the mainly difference of these vegetables tissues are used for food and shape, including root, stem, leaf and..
Also, have you found a report were says that BRC1 regulates flowering time?
Line 335
expressionED tan
Line 338
A lot of references are missing. Authors should explore more deeply the literature about TCPs and hormones.
Line 350, suggestion:
The OTHER TEN GENES rest containing the TCA element ten genes are upregulated
Line 351
directly regulated on the TCA element, but also causesMAY LINK other pathways to INDIRECTLY coordinately regulate the expression of the SOME TCP geneS.
Line 358
was not detectED


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: GENOME-WIDE IDENTIFICATION AND CHARACTERIZATION OF TCP FAMILY GENES IN BRASSICA JUNCEA VAR. TUMIDA
Review round: 1
Reviewer: 2

Output:
Basic reporting: MS needs to be screened for typo errors.
Experimental design: Experimental design is good. Some information needs to be mentioned in methodology. See my detailed comments in "comments to author" section
Validity of the findings: Fine but authors are suggested to make a putative mechanism diagram based on current investigation.
Additional comments: Manuscript by Jing He et al. is an interesting one, but before its further consideration, authors should amend below mentioned suggestions:
Comments/Suggestions
Line 19, use full botanical name at first appearance.
Line 19, “thirty-three” within a sentence use numerical values to denote numbers. Be uniform throughout the MS.
Line 26,mention full forms of abbreviations used at first appearance . for example “YA strains. After treatment with GA and SA”  Be uniform throughout MS.
Line 34-38, provide reference for specific parameter. For example : embryonic growth (reference), leaf
Development (reference), branching (reference), flowering (reference), circadian rhythm (reference), hormone signalling (reference),
Materials and Methods
Mention all growth conditions in detail.
What is the rationale behind selection of particular concentrations for hormones? Mention it.
What were qRT-PCR conditions?
Mention original reference for “The relative gene expression level was calculated using the 2-ΔΔCt method.”

Authors are suggested to make a  putative mechanism diagram on the basis of current results and discussion. Cite that figure in conclusion section. This figure will be very informative for readership.


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: GENOME-WIDE IDENTIFICATION AND CHARACTERIZATION OF TCP FAMILY GENES IN BRASSICA JUNCEA VAR. TUMIDA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors provide an improved revised manuscript. Overall, the revisions have clarified issues that now enabled a more informed evaluation of the manuscript. The authors have responded to my queries. They have largely addressed them convincingly yet important concerns.
Experimental design: The experimental design is ok now.
Validity of the findings: Authors now provides a better disccusion of their findings.
Additional comments: No additional comments


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND CHARACTERIZATION OF TCP FAMILY GENES IN BRASSICA JUNCEA VAR. TUMIDA
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comments
Additional comments: N/A


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

Input:
Title: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: This is a very interesting study that addresses an important gap in the literature: the effects of caffeine on exercise performance among females while using low caffeine doses. I have a few minor suggestions that would like to see addressed before acceptance.

Introduction:
Line 66: one narrative review that explored the effects of caffeine in resistance exercise provided these recommendations as well:
https://link.springer.com/article/10.1007/s40279-018-0997-y
This might be of interest to the authors as the review is quite recent. Otherwise, no comments on the Introduction. Well-written overall.

Methods:
Line 141: were the participants fasted for the testing sessions? Please specify the time of day at which the testing was performed and specify if the time of day was standardized for each participant.

It seems that the effectiveness of the blinding was not explored. If not, please add this as a study limitation, as shown by Saunders et al.
https://www.ncbi.nlm.nih.gov/pubmed/27882605

Discussion:
264: the mean effect size value is indeed a larger effect than what is previously reported in the meta-analyses that explored the effects of caffeine on strength. However, I think it would be worth noting that 95% CIs in the present study overlap with the 95% CIs in the meta-analyses that explored the effects of caffeine on strength.
https://www.ncbi.nlm.nih.gov/pubmed/20019636?dopt=Abstract
https://www.ncbi.nlm.nih.gov/pubmed/29527137?dopt=Abstract
Given the overlap of 95% CI, this might indicate the effect in the population is somewhat lower than the effect observed in this analyzed sample. I believe that adding discussion about these points is worthwhile.

Line 285: please reference the meta-analysis by Polito et al. here as well. https://www.sciencedirect.com/science/article/abs/pii/S0765159716000563

Line 300: please specify that RPE was assessed after completing the test and note that there is evidence supporting lower RPE following caffeine ingestion when assessed during exercise.
https://www.ncbi.nlm.nih.gov/pubmed/15773860

Limitations section
If the time of day was not standardized, this is another limitation worth specifying.

Figures:
Very nice figures, even though they might benefit from higher resolution.


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

Input:
Title: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 1
Reviewer: 2

Output:
Basic reporting: •	Clear, unambiguous, professional English language used throughout.
Yes.

•	Intro & background to show context.
It is well and clearly written. I do suggest to the authors to justify why women should have different response from caffeine consumption than men.

•	Literature well referenced & relevant.
Yes.

•	Structure conforms to PeerJ standards, discipline norm, or improved for clarity.
Yes.

•	Figures are relevant, high quality, well labelled & described.
In figures 1 to 4, change caffiene for caffeine.

•	Raw data supplied
Yes.
Experimental design: •	Original primary research within Scope of the journal.
Yes.

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

•	Rigorous investigation performed to a high technical & ethical standard.
Yes.

•	Methods described with sufficient detail & information to replicate.
Yes.
Validity of the findings: •	This study has higher degree of novelty and practical information.

•	This study showed a null effect of ~1.5 g/kg caffeine supplementation over RPE, RPD and TTF (regardless of load), and performance improvement for MVC. The audience should be aware about this information.

•	Meaningful replication encouraged where rationale & benefit to literature is clearly stated.
Yes.

•	All underlying data have been provided; they are robust, statistically sound, & controlled.
Yes.

•	Speculation is welcome, but should be identified as such.
Addressed.

•	Conclusions are well stated, linked to original research question & limited to supporting results.
Yes.
Additional comments: The authors did a great work with practical information.
I have minors concerns about this study, as follow:
•	Title: I do suggest withdrawing “…as well as muscular endurance…” because the statistical analysis did not support this statement;

•	Introduction: I do suggest including a sentence (or paragraph) justifying why women should have different outcomes than men;

•	Statistical analysis: Although it was a great and detailed work, the authors could improve the information for the audience including the homogeneity, normality and sphericity analysis;

•	Results: fix the figures (1-4) (change caffiene for caffeine);

•	Discussion: TTF – form line 279 to 283 – I do suggest deleting this statement. I understood the authors’ thought. However, the descriptive data has too large standard deviation which disagree with the speculation/analysis done. It will impact in the study’s title and the last sentence in the conclusion;

•	Conclusion: I do suggest deleting the last sentence “Since caffeine consumption can present with some deleterious side effects and withdrawal symptoms we suggest a low caffeine dose might be suitable to attain enhancements in strength and muscular endurance.” Because the data does not agree with it.


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

Input:
Title: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript presented aims to assess the potential ergogenic effects of acute low dose caffeine consumption on muscular strength in females. It is evident that the work has been meticulously conducted and the written communication is generally well presented however the research rationale and key findings need to be better contextualised with respect to the current state of the literature. The manuscript offers an important insight into a number of recently identified contemporary issues in this field by firstly examining the effects of a relatively low dose (200mg) caffeine (below the typically cited 3mg.kg threshold believed to induce an effect on physical performance) and by adding to the dearth in published articles examining the effect of caffeine supplementation in females.
Experimental design: The research question is clearly communicated and the experimental design appropriate to address the aims of the study. Some further work is needed to better contextualise the need to answer the question proposed and there are some minor amendments are required  to improve the clarity of the method. Please see specific comments below.
Validity of the findings: The data provided and the statistical approach is adequate with the discussion and conclusion well focused on the primary results. As with the introduction section, more work is needed to better contextualise these results and the broader application of these findings could be more explicitly presented. See specific comments below.
Additional comments: Abstract
-	In the method, please indicate what muscle group was tested at what angle the tests were performed. It would also be worth further indicating the characteristics of the population, (i.e. trained or untrained; habitual caffeine users?)
-	In the results section please be consistent in the number of decimal places when reporting values.
-	Lines 85-86. It is not clear if this sentence relates to the effect of caffeine or indicates a difference between the two tests? The effects of caffeine on 70% MVC are unclear.
-	Line 42. The sentence starting here is awkward, please reword.

Introduction
-	The first two paragraphs should be shortened and amalgamated into one to provide a more precise overview of the topics specifically related to the area.
-	I’m not sure that the argument around being a ‘considerable gap’ in literature with respect to exploring caffeine effects on muscular strength still holds given the number of recent systematic reviews on this subject. Would it not be better to state that despite such evidence there is a larger amount of conflicting findings for this mode of exercise and that caffeine effects are likely influenced by, participant characteristics, dose, exercise modality, muscle group?
-	Line 59 – is ‘side-effects’ the correct term here?
-	There needs to be a more robust and critical account of previous evidence exploring the effects of caffeine in females in order to better contextualise the need for this work. Most importantly there is a need to acknowledge the reasons for why there is limited work in females and the potential pitfalls in experimental design that may influence the results.
-	The section on line 67 needs some further thought. An optimal dose is likely to be individual and exercise specific. The rationale for examining low dose caffeine (and specifically 1.5mg.kg) needs to be more explicitly presented. The reviews by Pickering et al (https://doi.org/10.1016/j.nut.2019.06.016) and Spriet et al (https://doi.org/10.1007/s40279-014-0257-8) maybe useful.
-	Line 78 – this has been considered in an isolated muscle model (https://doi.org/10.1007/s12576-012-0247-20
-	Line 96 – it would appear that your study has a number of differences to the study cited. Consider removing this here and incorporating into the review of the literature above in order to improve clarity

Method
-	Line 113. It would be useful to cite the mean and standard deviation for the typical caffeine consumption to categorise the participant’s in to low or high habitual caffeine consumers,
-	Line 126 – What angle was the MVC completed at? Why was this chosen?
-	Did you measure or control for contraception/phase of the menstural cycle? If not this is a limitation of your study and the potential impact of this on the findings should be considered in further detail in the limitations section
-	Line 131 – given your dosing strategy I think you need to be clear in the abstract and at relevant parts throughout that you are actually examining the effects of 200mg of caffeine and that 1.5mg.kg is the average relative dose. In its current form this is a little misleading.
-	Line 148 and 149 appear to be a repeat
-	Was MVC assessed during all four experimental trails? Is so it is not clear how this was analysed in the outline of the statistical method
-	Line 193 – please provide a reference to support the thresholds used to denote large and moderate effect sizes

Results
-	The figures are well presented and it is nice to see that you have opted to report the individual reposes. Please check that the quality of the images aligns to the requirements of the journal.

Discussion
-	Line 242 replace ‘and’ with ‘an’, add to this first paragraph by providing a summary of the key findings
-	The paragraph starting on line 246 seems a little out of context, a more concise version of this would be useful in the introduction to better justify the dose. Also consider are the examples provided relevant. Is it likely that athletes are going to consume a Starbucks prior to physical activity?
-	The discussion of MVC data needs to be better contextualised with respect to previous work examining the effect of caffeine on strength and caffeine effects in female participants
-	TTF – please outline how these results compare to other studies examining the effect of caffeine on repeated muscular contractions
-	If perception of effort/pain did not account for the results presented, can the authors suggest what the likely underlying mechanism(s) may be.
-	A more explicitly account of the applications of this data for practice would be useful
-	Conclusion – the negative side effects of caffeine consumption should be discussed earlier in the manuscript to better contextualise the application of the data presented


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

Input:
Title: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Great work with the revision. Congrats on this 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: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: I am pleased with the article's improvement. Nothing to add.


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

Input:
Title: A LOW CAFFEINE DOSE IMPROVES MAXIMAL STRENGTH, BUT NOT RELATIVE MUSCULAR ENDURANCE IN EITHER HEAVIER-OR LIGHTER-LOADS, OR PERCEPTIONS OF EFFORT OR DISCOMFORT AT TASK FAILURE IN FEMALES
Review round: 2
Reviewer: 3

Output:
Basic reporting: Please see previous review.
Experimental design: Please see previous review.
Validity of the findings: Please see previous review.
Additional comments: The authors have adequately addressed the comments outlined in my original review.


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

Input:
Title: INVENTORY STATISTICS MEET BIG DATA: COMPLICATIONS FOR ESTIMATING NUMBERS OF SPECIES
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this manuscript the authors draw attention to the effects of small errors in biodiversity databases on the species richness estimations generated by means the use of nonparametric estimators. The cited literature  is up to date and the purpose is clear.
Experimental design: This study is in fact a cautionary note aimed to highlight the dramatic effects in these estimations when occurrence data are managed uncaringly, containing taxonomic or occurrence errors. Using simulated assemblages and introducing a 10% of erroneous rare species, the authors demonstrate that the appearance of these species causes overestimations when the number of replicates increases. The rationale and the provided results are clear and the message that is sent to the users is useful
Validity of the findings: The message of this study could be yet more useful if different numbers of “error” species are included in the analyses (e.g. 1%, 3%, 5% and 10%) Please consider that in the example provided in the figure 1 only 3% of the species are erroneous. Additionally, I also suggest that the results and the figures 3 and 4 should include the mean number of species estimated when these rare species are introduced, also providing the 95% confidence interval. This may contribute to better understand the real contribution of these erroneous occurrences in species richness estimations.
Additional comments: I consider that this manuscript can be a nice and useful piece of information for all people interested in managing the data provided by biodiversity databases. I would encourage the authors to extend your results.


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: INVENTORY STATISTICS MEET BIG DATA: COMPLICATIONS FOR ESTIMATING NUMBERS OF SPECIES
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper explores the effects of very rare occurrences in large datasets on estimates of species richness through a set of simulations. As often-used estimates rely on the distribution of either abundances in a sample or frequencies of incidence throughout sample sets, the analysis is relevant in the field, for it provides a warning about a potential source of overestimations of richness. The authors have followed a sensible, classically empirical analytical path: let’s create an artificial assemblage, sample it as if it was real, and observe how critical parameters (singletons and doubletons in incidence-based estimates) vary with sample size, and then introduce very rare incidences and compare whether the estimates hold or not.
The paper is correctly written, well referenced and structured, and presents the findings in a clear way. However, the Methodology section needs some clarifications.
I have no issue on the design of the experiment, and find it elegant and conceptually reproducible. In fact, I have tried to reproduce some of it, using a different, independent set of tools, to confirm its findings, and was able to get to similar results but only under somewhat different conditions: namely, with uniform or normal distributions for the samples, or, if using lognormal distributions as described in the paper, only for much larger samples.
I believe that the reason I have not been able to fully verify the results may not lie in the original experiment, but, rather, on how it is explained in the paper and therefore my ability to fully grasp all the details. When designing my own simulations, I struggled to find some parameters I think were missing. If they were not necessary, then I did not understand some important point. In either case, the paper needs some clarification. Very likely these clarifications may enable much better reproducibility. Then, whether the results hold or not will be up to the readers repeating the experiment as intended by the authors.
Experimental design: The first, main issue is the definition of “samples”, “replicates”, and “days”. They seem to be used interchangeably but in a rather confusing way. For example, all figure footings refer to “sample days” while the main text refers to “samples”, but the context seems to indicate that sampling days and (part of the) samples are replicates. Throughout the paper there is an inconsistency abut what a sample is. My interpretation (after several reads and examination of figure 1) is that a “sample” in this context is a set of 5, 7, 10… inventories, taken on a “day” each; that each inventory contains a set of incidences of [up to] 100 species; that the contingency matrix that condenses the samplings can therefore be 5, 7, 10… columns wide, and that each of these matrices has been redrawn up to 100 times. It is paramount to describe the simulations in good detail, and that the figure footings are consistent with the main text.
The second important issue (or not?) is that the “sample size” seems to refer to the number of inventories in the sample, which is correct for incidence data. However, a simulation of incidence data requires another parameter: What are the sizes of the populations being sampled within the assemblage? The paper states that the assemblage is log-normally distributed and the distribution parameters are supplied, which enables us to calculate the relative sizes of each population (=species) and therefore the probability of occurrence in the inventory relative to other populations in the assemblage. Yet, whether a species is “sampled” (i.e., appears in an inventory) depends on that relative probability but also on the sample size: number of individuals in the inventory. By definition, a species occurs in an inventory if at least one single individual is sampled (n=1), and all species being sampled in just one inventory within a multiple (=multi-inventory) sampling contribute to Q1; but the parameters of the lognormal distribution, which are the average and standard distribution of ln(x), are given in the text (or so I believe) in relative terms, despite being presented as average abundance. In order to create the assemblage, the probabilities must be applied to a certain population size. For a 100-species assemblage following a lognor distribution with average abundance = 1.5 and a standard deviation = 2, a heap of species would be represented by a fractional individual, which is unrealistic. Thus, an overall abundance (assemblage size) should be given ensuring that the 100-species complete inventory includes at least one individual of the least-represented species.
When using such inventory sizes, simulations seem to show that Q1 and Q2 do not saturate as described in the paper but take longer, unless the underlying distribution is much more uniform.
Validity of the findings: See comments above.
Additional comments: Other lesser issues on the paper can be easily corrected, and are annotated in the PDF. For example, (line 82) the authors have explored four nonparametric estimates (not three as written). There is also a panel missing in figure 4 (footing says 3 and there are only 2).
My recommendation is that the paper will be publishable when authors clarify the details of their simulation enough for other researchers to be able to reproduce the results without second guesses. A good rewrite of the Methodology would be highly convenient. Indeed, supplying a sample matrix of the artificial assemblage and a worked-out example of the simulation would be quite useful, in addition to the already-supplied scripts. These alone do not seem enough to fully and reliably reproduce the reported results.


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: INVENTORY STATISTICS MEET BIG DATA: COMPLICATIONS FOR ESTIMATING NUMBERS OF SPECIES
Review round: 2
Reviewer: 1

Output:
Basic reporting: In my former suggestions I recommended including different numbers of “error” species, but the authors reject this suggestion because it “will come to the same conclusion as those that we have already presented, and will not reveal any new properties”. This is probably true, although could improve understanding the linear or non-linear character of the relationship between the number of these errors and the overestimations. Be as it may be, as the own authors state, the results of this manuscript “demonstrate something that is entirely obvious”; i.e. that including rare species as consequence of erroneous occurrences or taxonomical identifications may provide erroneous estimations. Thus, taking into account that this study must be seen as a cautionary note, I accept the current version of the manuscript expecting that it improve the management by users of the biodiversity information with non-parametric estimators.
Experimental design: I consider that the provided methods are adequate
Validity of the findings: I consider that this manuscript can be a nice and useful piece of information for all people interested in managing the data provided by biodiversity databases
Additional comments: anyone


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: NONALCOHOLIC FATTY LIVER DISEASE, A POTENTIAL RISK FACTOR OF NON-SPECIFIC ST-T SEGMENT CHANGES: DATA FROM A CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. Well-structured and well written manuscript with clear and professional English language.
2. Professional and well conducted tables and figures.
3. Clearly explained introduction explaining the clinical importance of the topic.
Experimental design: 1. Well defined and relevant research question with very limited published research on the topic.
2. Clearly described methods that allow it to be reproducible by other researchers.
Validity of the findings: 1. Although this isn't the first study to evaluate the association between NAFLD and nonspecific ST-T segment changes, it is the largest in population size which adds to the current literature.
2. Multivariable logistic regression analysis was performed to evaluate for confounding factors.
3. Conclusions can't confirm causality (as mentioned by the authors). Nevertheless, the reported conclusions were based on the demonstrated results.
Additional comments: -	 Main theme of the manuscript:
This is a retrospective cross-sectional study conducted on 32,922 participants who presented in a single hospital (Wuhan Union Hospital), aged between 18 to 65 years. The aim of the study was to assess the association between NAFLD and nonspecific ST-T segment changes on ECG. The authors demonstrated that subjects with NAFLD are independently associated with an increased incidence of ST-T segment changes on ECG, even after adjusting for potential confounders.

-	Current literature:
The current literature is limited in data evaluating the association between NAFLD and non-specific ST-T segment changes on ECG. Two studies and a systematic review evaluated this association. A cross-sectional study conducted on Korean adult subjects by Lee et al. involving 50 NAFLD patients and 100 age and gender matched controls demonstrated that ischemic heart disease changes on ECG weren’t independently associated with hepatic steatosis. On the other hand, Vendhan et al. conducted a cross-sectional study on 541 participants demonstrating a significant association between nonobese NAFLD and coronary artery disease defined by ECG changes including ST-segment depression, Q-wave changes, or T-wave changes associated with ischemic heart disease, even after performing multiple logistic regression analysis. Moreover, a recently published systematic review conducted by Ismaiel et al. evaluated the methodological quality of the previously mentioned studies and rated the first mentioned study (Lee et al.) as “fair” and the second (Vendhan et al.) as “good”, supporting the presence of ECG changes related to ischemic heart disease in NAFLD.

-	What’s new?
The current study is of interest in the fields of gastroenterology, cardiology as well as internal medicine as NAFLD is a multisystem disease with a rapid increase in prevalence worldwide. Cardiovascular risk stratification in NAFLD patients using an ECG, a simple and widely used device, is of great clinical significance. The authors conducted a study with the largest study population at the moment assessing nonspecific ST-T segment changes in NAFLD allowing a better evaluation of this association on a large number of subjects. Demonstrating an increased incidence of ST-T segment changes in NAFLD patients, even after adjusting for confounders on such study population adds to the current literature as results can be generalized to a larger extent.

-	Comments:
1.	Please mention the period of recruitment in which the subjects were reviewed at the physical examination centre in Wuhan Union.
2.	Please describe the characteristics of study participants (eg demographic, clinical, social).
3.	You mentioned that patients with missing liver function tests were excluded from the study. I couldn’t find AST, ALT, GGT and ALP levels? Were they assessed in the included study participants?
4.	In the current study, hepatic steatosis severity evaluated using abdominal ultrasonography was classified into normal, mild, or moderate/severe although the ultrasonographic severity classification of hepatic steatosis is divided in to grades 0-4 (grade 0, absent; grade 1, mild; grade 2, moderate; grade 3, severe). Why did you combine moderate and severe together in the same severity grade?
5.	The discussion section didn’t discuss the current results of the published studies evaluating nonspecific ST-T segment changes in NAFLD. Please have a look at the currently published studies evaluating this association and discuss how your results are similar/different. Below are some studies evaluating this association.
•	Study 1 (2006):
(https://www.tandfonline.com/doi/full/10.1080/00365520500319591?scroll=top&needAccess=true)
•	Study 2 (2014):
(https://www.liebertpub.com/doi/10.1089/dia.2013.0165)
•	Systematic review (2019):
(https://www.jgld.ro/jgld/index.php/jgld/article/view/344/168)
6.	How would you explain that patients with mild hepatic steatosis have an increased incidence of nonspecific ST-T segment changes compared to patients with moderate/severe steatosis? You can discuss this in the discussion section.
7.	The word “Notably” in the second line of the discussion section should have the letter n in lowercase.
8.	Use of the abbreviation “NAFLD” isn’t consistent throughout the manuscript. Some sentences have the abbreviation while others have the full word mentioned (nonalcoholic fatty liver disease). Please mention the full word with the abbreviation on the first appearance in the manuscript and then use the abbreviation only.

-	Limitations:
1.	Diagnosing NAFLD using abdominal ultrasonography might underestimate the prevalence of NAFLD.
2.	The retrospective cross-sectional study design doesn’t allow the demonstration of causality between NAFLD and nonspecific ST-T segment changes in NAFLD.
3.	Residual confounders were not excluded due to the measurement errors of these variables.

-	Strengths:
1.	Well-structured and well written manuscript with clear and professional English language.
2.	Professional and well conducted tables and figures.
3.	Limited literature published on this topic (2 cross-sectional studies and 1 systematic review published on PubMed) making it of significant clinical importance due to the associated cardiovascular risk.
4.	Good methodological quality evaluated using STROBE quality assessment tool for cross-sectional studies.
5.	Large sample size.

-	Summary:
I believe that this is a well conducted cross-sectional study that could be considered for publication after performing the above-mentioned modifications that will improve the quality of 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: NONALCOHOLIC FATTY LIVER DISEASE, A POTENTIAL RISK FACTOR OF NON-SPECIFIC ST-T SEGMENT CHANGES: DATA FROM A CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well written in professional English.
There are only few parts I would modify:
-	I would replace the word ‘coincidentally’ in line 50 with a more straightforward word;
-	I would change the word “obese” in line 141 with “heavier” (or similar), seen that the average BMI was way inferior to 30;
-	In lines 149 and 152 the phrase “the proportion of NST changes” could be misleading; I would change it with “the proportion of patients with NST changes”;
-	I would integrate the phrase in line 139-140 as it follows: “a higher prevalence presented in middle-aged and older individuals (third and fourth quartile of age)”. I think it gives a better link to your study’s population.

I think that the introduction needs more details. I personally would add some more background information about NAFLD in lines 49-50 (see Targher G et al. NEJM, 2010).

The structure is conform to PeerJ standards.

Figures are relevant and of high quality.
In Table 2, line 4 I would change the word “unadjustment” with “unadjusted”

Raw data are supplied and are satisfactory.
Experimental design: This is an original primary research. There is no such a research in literature yet. On Medline we can find correlations between NAFDL and QTc prolongation, PR prolongation, presence of AF, ventricular arrhythmias, RBBB, but none with non specific ST-T segment changes.
This is the reason why I think the research fills an identified knowledge gap

The investigation is rigorous. I think there may have been a mistake about the numbers of the participants included in the final analysis (32922 as shown in the fig. 1 vs 32864 as described in line 91?). The raw data show 32922 participants.

Methods are described in detail.
Validity of the findings: I do appreciate that researchers encourage further studies on the subject

All the data are provided and look statistically sound

Speculation should be identified as such: in line 180 I would change the verb “indicate” with the verb “suggested”, only to underline once more that correlation is not causation

Conclusions are well stated
Additional comments: I think the article is well written and it is an original research.


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: NONALCOHOLIC FATTY LIVER DISEASE, A POTENTIAL RISK FACTOR OF NON-SPECIFIC ST-T SEGMENT CHANGES: DATA FROM A CROSS-SECTIONAL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: 1. Well-structured and well written manuscript with clear and professional English language.
2. Professional and well conducted tables and figures.
3. Clearly explained introduction explaining the clinical importance of the topic.
Experimental design: 1. Well defined and relevant research question with very limited published research on the topic.
2. Clearly described methods that allow it to be reproducible by other researchers.
Validity of the findings: 1. This study evaluates the association between NAFLD and nonspecific ST-T segment changes with the largest study sample which adds to the current literature.
2. Multivariable logistic regression analysis was performed to evaluate for confounding factors.
3. Conclusions can't confirm causality (as mentioned by the authors). Nevertheless, the reported conclusions were based on the demonstrated results.
Additional comments: The authors modified the manuscript as per the previously mentioned suggestions. This is a well conducted cross-sectional study that adds new insights to the scientific literature. I believe that the manuscript can now be considered for publication.


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

Input:
Title: NONALCOHOLIC FATTY LIVER DISEASE, A POTENTIAL RISK FACTOR OF NON-SPECIFIC ST-T SEGMENT CHANGES: DATA FROM A CROSS-SECTIONAL STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The article is well written in professional English, there are only few things I would change:
-	Line 200: the present study (not studies) provided evidence;
-	Line 205: I would change the phrase “the population we surveyed was relative health patients“ with “the population we studied was made up of relatively healthy people”
Introduction and background are well written; literature is well referenced and relevant.
Relevant figure, adequate and well labelled
Raw data: satisfactory
Experimental design: As I stated on the previous review I think this is an original primary research. On Medline several correlations are described between NAFDL and cardio-vascular disease, but none specifically with NST changes.
I think this research fills a knowledge gap.
The investigation is rigorous; the methods are described in detail
Validity of the findings: Data are provided and look statistically sound
Speculation is identified as such.
Conclusions are well stated.
Researchers encourage further studies on the subject.
Additional comments: I do appreciate the changes that have been made, which in my opinion make the article more complete. I do appreciate the clarification on the approval to the study given by the ethics committee (and the file provided).
I think the article is well written and it is an original research.


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

Input:
Title: THE BIOMARKERS OF KEY MIRNAS AND TARGET GENES ASSOCIATED WITH ACUTE MYOCARDIAL INFARCTION
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. The text of the manuscript must be corrected by a native English speaker. The current title of the manuscript is not easily understandable.
2. Literature is relevant and well referenced.
3. Figures and Tables are relevant to the content of the article, and appropriately described and labeled.
4. Raw data is available in accordance with the Data Sharing policy.
5. Abbreviations, when first used, should be explained in the text as well as in a separate section.
Experimental design: 1. The research topic of study is not novel. Numerous studies have been undertaken to identify and understand miRNA/mRNA biomarkers in acute myocardial infarction.
2. The aim of the study should be more emphasized in the text of the manuscript.
3. The number of analyzed samples acquired from the GEO database (normal and AMI groups) should be provided in the text of the manuscript.
4. The text of the paragraph “Analyses of miRNA-mRNA targets” should be more detailed.
5. Full names of the programs used for data preprocessing and expression data analysis should be provided.
6. The rationale for the selected dosage of doxorubicin should be clearly explained. Did the Authors test any other doses of doxorubicin?
7. More detailed demographic and clinical characteristics of the AMI patients could give valuable information.
8. What is the source of the investigated miRNA from the AMI patients?
9. U6 is not a suitable reference control for the quantification of circulating miRNAs. U6 is not miRNA, and does not reflect the biochemical character of miRNA molecules in terms of their transcription, processing and tissue-specific expression patterns. Were some other endogenous miRNA controls tested as a reference control?
10. The sequences of reverse primers for the verified miRNA should be provided.
11. More detailed description of the RT-qPCR statistical data analysis and ROC curves should be introduced into the text of manuscript.
12. It is not clear if the RT-qPCR experiments were performed using only H9C2 cell line or also with the AMI patients’ blood samples?
13. According to the MIQE Guidelines (Bustin el al., 2009), the abbreviation RT-qPCR (reverse transcription quantitative real-time PCR) should be used in the context of quantitative RT-PCR.
14. It is not clear what kind of data was used to construct the ROC curves.
Validity of the findings: 1. All underlying data has been provided; it is robust, statistically sound, and controlled.
2. Conclusions are well stated, linked to the original research question and limited to supporting results. The limitations of the study are pointed out.
Additional comments: No additional comments


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

Input:
Title: THE BIOMARKERS OF KEY MIRNAS AND TARGET GENES ASSOCIATED WITH ACUTE MYOCARDIAL INFARCTION
Review round: 1
Reviewer: 2

Output:
Basic reporting: Some well known biomarkers are  use for diagnosis of acute myocardial infarction,  but identification of new biomarker can improve effective treatment for better prognosis of AMI patients. The aim of this study was to investigate miRNAs expression in AMI patients and identification of  miRNAs as potential diagnostic biomarkers for AMI patients. To do this, the authors used microarrays results deposited in the Gene Expression Omnibus by two independent groups. Next, the authors performed bioinformatics analysis in order to find  the association of particular miRNAs with mRNAs. Also the H9C2 cell and samples from AMI patients were used for clinical evaluation of the identified miRNAs and their target genes. The English is clear and acceptable.
Experimental design: The authors do not provide row data for qPCR experiments. Also there is no information who was responsible for this experiments (see “Authors’ contributions”).
2.)	There is no baseline characteristics of MI patients and control group (sex, age, clinical parameters etc.) The authors do not give results of miRNAs expression in blood samples patients. Whether they were consistent with the results obtained for the tissue cultures?
3.)	In case of experiments performed on the cell lines there is a lack of access to methodological details, raw data, number of replicates etc.
4.)	Authors state that “miR-139-3p, miR-24-1*, miR-598 and miR-142-3p, have not 259 been linked to the regulation of cardiac performance” what is not entirely true because all of them were mentioned in articles connected with cardiovascular processes:
-	miR-139-3p  Sci Rep. 2017 Aug 21;7(1):8898.
-	miR-24-1 - Circ Heart Fail. 2019 Mar;12(3):e005765.
-	miR-598 - Eur J Heart Fail. 2013 Oct;15(10):1138-47.
-	miR-142-3p–PloSOne.2017 Jan 26;12(1):e0170842.; Biomed Pharmacother. 2018 Dec;108:1347-1356; J Mol Cell Cardiol. 2019 Aug;133:12-25.
Validity of the findings: A number of previous studies have already shown that plasma miRNAs expression changes in patients in the acute phase of a myocardial infarction. While the association of particular miRNAs with MI was suggested in this work, evidence is lacking in supporting it being a prognostic biomarker.
Additional comments: No additional comments


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

Input:
Title: THE BIOMARKERS OF KEY MIRNAS AND TARGET GENES ASSOCIATED WITH ACUTE MYOCARDIAL INFARCTION
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In their manuscript entitled, ‘The biomarkers of key miRNAs and target genes associated with acute myocardial infarction,’ Wang et al. explored miRNA expression in cell culture and clinical samples of myocardial infarction (MI) and controls in order to identify candidate markers for the diagnosis of acute MI

The manuscript seems to attempt to address a need for more sensitive biomarkers for the detection of MI.
However, it does not become clear why there is a need for new biomarkers in the first place. Furthermore, the manuscript is not providing structured information as to exactly which types of experiments were done at which time point and in which sample types. Most importantly, it remains unclear what the overall intention of the manuscript is – discovery of novel circulating biomarkers for MI? discovery of novel miRNA-related signalling pathways?

1) It does not become clear what the intention of the manuscript is. Is it to identify novel circulating biomarkers for a potential clinical application in the diagnosis of acute MI or is it to investigate miRNA pathways on a cardiac cellular level?

2) It does not become clear what was done in the manuscript:
-	Lack of information on the type of samples used
-	Lack of information on which samples (cell culture, blood (which blood fraction – whole blood? Blood cells?)) were used for which analysis
-	Which sample/cell type was “Identification of DE-miRNAs”, “MiRNA – target gene interactions”, “Enrichment analyses of the target genes”, Protein-protein interaction network”, “Biological analysis of the hub genes” performed in?

3) qPCR-based validation was performed in clinical samples in MI patients vs. controls. The patients need to be characterised  baseline characteristics (i.e. age, gender, cardiovascular risk factors, medication). Especially anti-platelet medication is of importance since this alters platelet-derived miRNAs.

4) Immensely important is the application of heparin in patients/individuals before blood was drawn, since heparin is a strong confounder of qPCR-based DNA/RNA detection methods. Were any efforts undertaken to overcome the heparin bias (heparinase treatment?)

5) With qPCR-based RNA quantification, normalisation is of utmost importance. How was the discovery of normalizers carried out and which normalizers were used? How were the qPCR raw data transferred into relative quantity? Delta-delta Cq method?

6) The AUC analyses need to be compared with established biomarkers for AMI (i.e. troponins). Otherwise the results cannot be interpreted properly.

7) In the abstract it does not become clear why there is a need for additional diagnostic biomarker for AMI. It is widely known that cardiac troponins can be detected highly sensitive and at the same time show high specificity. The broad international audience might want to learn about the necessity to explore alternative biomarkers.
In line with this, the authors state in line 60 “…high mortality of AMI is its late diagnosis, which is induced by the lack of highly sensitive and specific biological markers.” In the light of the highly sensitive and specific cardiac troponin biomarkers, this sentence is wrong and needs to be amended; the current ‘problem’ with high sensitive cardiac troponin is rather the high sensitivity which comes along with more patients being evaluated in the emergency department before AMI can be ruled out and thus causing higher socio-economic costs.

8) Furthermore, beginning in the abstract the authors should specify what type of samples were used.

9) As per the above point 4, please specify in the method section how blood was taken, which blood collection tubes were used (RNA-protective reagents such as PAXGene or EDTA etc.?). This is particularly important since upon freezing blood cells will lose their integrity and release their content into the cell free compartment. After thawing this released DNA and or RNA is prone to degradation.

10) The term ‘normal samples’ most likely refers to control samples I suppose? I suggest to use the latter term.


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

Input:
Title: THE BIOMARKERS OF KEY MIRNAS AND TARGET GENES ASSOCIATED WITH ACUTE MYOCARDIAL INFARCTION
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
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: EXTENT, PATTERNS AND DEMOGRAPHIC CORRELATES FOR PHYSICAL FIGHTING AMONG SCHOOL-ATTENDING ADOLESCENTS IN NAMIBIA: EXAMINATION OF THE 2013 GLOBAL SCHOOL-BASED HEALTH SURVEY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Abstract:
1)	Line 55: Please rephrase this sentence, as all and any physical fighting should be referred to as problematic.
Introduction:

2)	Lines 67-71: Please expand this by adding information on morbidity and mortality from physical fighting in adolescents. Also, more recent references should be added.

3)	Line 78: Please reconsider the use of phrase ‘’problematic physical fighting’’, as it might suggest that there is one which is not problematic, or precisely define the phrase. Does this refer to 2 or more fights?

4)	Introduction fails to clearly indicate the knowledge gap that authors aim to address. Add information on prevalence estimates of physical fighting in the world and in Namibia, if any. Add information on general risk factors for such behavior, and any disagreement in results of previous studies on these risk factors.

5)	Lines 76-80: It is not clear how this study builds upon a previous study which was conducted in the US. Please check the reference you used.

Methods:
6)	How were missing variables handled?

7)	At which level was significance considered in the bivariate model, in order for a variable to be included in the second model?

8)	You stated in the abstract, but not in methodology, that factors associated with physical fighting were identified by a literature search. There is no mention of this in the section Methods. Why does the list of independent variables not include factors which literature recognizes as risk factors for physical fighting in adolescents, such as substance abuse, smoking, socioeconomic factors, etc.?

Results:
9) Lines 154-155: The text ‘’as they did not include ‘1’ in their 99% CIs.’’ Is redundant.

Discussion:
10)	Lines 173-175: The cited reference is not conducted in Namibia.

11)	Lines 192-197: The discussion of risk factors which you found to be statistically significant should be expanded by comparing your results with other studies.

12)	Lines 194-197: This sentence needs to be checked, and rephrased, as it states truancy is associated with truancy. Previous studies have showed that increasing number of joint adverse behaviors is associated with increasing odds for physical fighting (‘’Physical fighting and associated factors among adolescents aged 13–15 years in six western Pacific countries’’ 2017, by Yang et al.).

13)	Please consider briefly discussing the protective effect of loneliness, as some of the identified risk factors (e.g. being bullied, anxiety, having a suicidal plan) can be associated with loneliness.

14)	Lines 229-234: Information on potential risk factors could be missing too.

15)	A limitation is also comparability, given the use of different self-reported measures, definitions of variables, etc.
Experimental design: See above.
Validity of the findings: See above.
Additional comments: See above.


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: EXTENT, PATTERNS AND DEMOGRAPHIC CORRELATES FOR PHYSICAL FIGHTING AMONG SCHOOL-ATTENDING ADOLESCENTS IN NAMIBIA: EXAMINATION OF THE 2013 GLOBAL SCHOOL-BASED HEALTH SURVEY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article meets the standard. However could be strengthened, especially the Introduction. It would be strengthened by addressing changes in physical fighting between the two survey periods.
Line 73, is not true. There is information on the prevalence and general risk factors.

Methods' section
Under setting, describe the age structure of the population as well as the enrolment rates for the study population.

In a few areas, present tense was used instead of past tense. e.g. line 99.."has"
line 110 on definition involvement in a physical fight. The authors chose "2 or more fights" which in some studies "1 or more" was used. Care should be taken when comparing the findings with other findings.

Statistical analysis
what is the benefit of applying Rao-Scott Chi-square test and the design adjusted t test wen the analysis was weighted? Compare the results of these analyses and choose to report more accurate estimates.

Results
line 135: avoid starting sentences with digits
line 147: Results from Table 4 are different from the ones in the paragraph starting in this line
line 152: There is no point to test at 99% confidence level as well.
line 155: no need to give a lecturer on the interpretation of the results of the confidence interval

Discussion
line 173: as the authors stated, the two studies used different definition for physical fighting, hence, it is not worthy to go ahead and make the comparisons.
line 181: write HDI in full.
lines 209-213: this section is repeating paragraph 2 of the Discussion section.
lines 23-219 should be moved to the Introduction as part of the justification of the study.

References
An example of style of referencing from one of your recent articles has the following example:
Albarqouni S, Baur C, Achilles F, Belagiannis V, Demirci S, Navab N. 2016. AggNet: deep learning from crowds for mitosis detection in breast cancer histology images. IEEE Transactions on Medical Imaging 35(5):1313-1321

Clearly if the above format of references is the journals recommended style, then more work is needed to clean up the references. No websites should be indicated.
Experimental design: This is not an experiment.
Validity of the findings: The validity of findings is questionable because the figures in the text do not match those in Table 4.
Additional comments: No additional comments


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

Input:
Title: EXTENT, PATTERNS AND DEMOGRAPHIC CORRELATES FOR PHYSICAL FIGHTING AMONG SCHOOL-ATTENDING ADOLESCENTS IN NAMIBIA: EXAMINATION OF THE 2013 GLOBAL SCHOOL-BASED HEALTH SURVEY
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: A well argued 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: EEG BASED ASSESSMENT OF STRESS IN HORSES: A PILOT STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: Review of “EEG based assessment of stress in horses: A Pilot Study” by de Camp et al. submitted to PeerJ is an interesting and refreshingly new take on non-invasive assessment of mental states of horses in anticipation of stressful condition.

In this study, the authors make a compelling case for need to establish species specific assessment criteria for their mental wellbeing across animals such as horses. They also argue in favor of developing novel techniques and adapting available technologies for this purpose. In this pilot study, they make the case by their innovative use of telemetric 3 lead EEG recording and attempt to correlate it with a well-established behavioral measure called, horse grimace scale (HGS). The EEG recordings and HGS assessments were made under conditions of “rest”, when the horses were in their stables or home environment, contrasted with “stress” condition, when they were taken for medical treatment. Their findings, though preliminary, show that there was significant change in HGS and cross-frequency coupling in EEG signal between the two conditions. As a proof of concept, while their results should be taken with caution, still provide promising trend that should be easily verifiable and may lead to significant insights into physiological read outs of the internal states of horses.

The experiments and data were collected following adequate protocols and care. Though they lack technical sophistication of standard physiological recordings, it is worth noting that these were done during free behaving condition and under minimal or no restraint to the animal, which could be quite challenging.

Here are a few questions and comments for the authors to help improve the manuscript:
1.	Please provide rationale for choosing right somatosensory region for EEG recordings? Will the results be different if any other region on the head was selected for recordings?
2.	Under EEG recording section, authors mentions “Selected sequences lasted on an average 90s in case of the resting condition and 50 s in case of the stress condition.. “  - How many such sequences occurred in a typical recording session? How many times the same individual was recorded under two conditions?
3.	Any speculation with regard to the changes they saw in HGS and PAC, whether they were due to treatment stress or change in environment e.g. clinic vs home environment (change in local environment) OR treatment anticipation stress vs resting state.
4.	 Authors should discuss EEG changes more specifically that are related to their own data, for instance - a) their interpretation of the observed differential coupling strength between rest and stressful condition during up and down phases? b) intriguing but potentially interesting finding of a “progressive change in coupling strength” around ~8Hz and gamma band, rather than an abrupt change?
5.	In future studies it will also be interesting to relate cross-frequency coupling changes in low and high gamma band using multielectrode EEG with change in heart rate of the animal in different conditions of stress.

Minor points:
Remove commas and replace with standard decimal notations when providing data and statistics – such as p=0,0006 should be changed to p=0.0006. This should be done throughout 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: EZH2/H3K27ME3 AND PHOSPHORYLATED EZH2 PREDICT CHEMOTHERAPY RESPONSE AND PROGNOSIS IN OVARIAN CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This manuscript by Sun et al. investigated the correlation between the expression level of EZH2, pS21-EZH2, H3K27Me3 or pAkt and clinical behaviors of ovarian cancer. The data is clearly presented and the manuscript is well written. My primary concern is that different histologic subtypes of ovarian cancers arise from the different cells of origins and should be considered as distinct diseases. As the majority of the cases investigated in this study (53/65) were serous ovarian cancer, it is recommended that the authors should do analysis within the serous ovarian cancer and discuss the limitation of not a sufficient number of cases of other subtypes being included.

1. Fig2 and Fig1 used identical representative images for EZH2 and pEZH2. The images in Fig 2 provides little values and can be removed. Instead, the author should provide a correlation plot for EZH2 and chemo response. Given that both low pS21-EZH2 and low EZH2/lowH3K27Me3 correlated with better chemo-responses and survival, it will be interesting to analyze whether there is any correlation between pS21-EZH2 and EZH2 levels.
2. In Table 2, the combination of pAkt and pS21-EZH2 did not provide better value in predicting response to chemotherapy than pS21-EZH2 alone. The authors should comment on this in the discussion.
3. Line 169, the p-value is inconsistent with the value in Table 3.
4. Line 201. the word 'reprogram' is a verb and should be changed to a noun, reprogramming.


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: EZH2/H3K27ME3 AND PHOSPHORYLATED EZH2 PREDICT CHEMOTHERAPY RESPONSE AND PROGNOSIS IN OVARIAN CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript was written adequately, however it requires editing to correct some language errors.
Experimental design: Some important details are missing in Materials and Methods (see General Comments to the Authors).
Validity of the findings: The Discussion contains some speculation, which would be hard to avoid considering the descriptive and associative nature of the study.
Additional comments: Sun and collaborators determined the associations between the EZH2 canonical and non-canonical signaling pathways with chemotherapy response and prognosis in ovarian cancer. For this, the authors measured levels of EZH2, H3K27me3, pAKT1, and pS21EZH2 by IHC in ovarian cancer tumors from 65 patients. The authors reported that EZH2/H3K27Me3 and pS21EZH2 IHC scores have predictive value for chemotherapy response and progression-free survival in ovarian cancer. Below are suggestions for improving the manuscript.

Comments
1.	Indicate when the tumor samples were collected.
2.	Lines 113-117. Provide the catalog numbers of the antibodies. Indicate what negative controls were included in the IHC runs.
3.	Line 128. IHC scores (line 122), not protein expression, were used for determination of associations.  IHC scores should be used throughout the manuscript when referring to protein levels in the tumors.
4.	Lines 144-145. Indicate the p-values of the correlations to make designations of “closely” and “poorly” correlated less arbitrary.
5.	Lines 154-155. When describing EZH2/H3K27Me3 or pAKT1/pS21EZH2 association with chemotherapy response, it is not clear if it is for each protein, either one, or both combined.
6.	For consistency use either chemotherapy resistance or chemotherapy response throughout the manuscript.
7.	Lines 197-198. Sentence is not clear. Please revise.
8.	The Discussion should be expanded to include possible mechanisms of EZH2 regulation of  response to platinum chemotherapy. Also, limitations of the study given the relatively small numbers of patients, and grouping of the samples only into serous and non-serous should be mentioned.
9.	Figure 1. Pictures of low staining for p-EZH2, p-AKT1, and EZH2 show no staining. These pictures should be replaced with representative pictures that actually show some staining. Were the correlations actually calculated using percentages of positive cells as indicated on the figure legend? If percentages were in fact used justify why it was preferred over IHC scores.
10.	Figure 2. Indicate how many patients were resistant or sensitive.
11.	Table 1 and Table 2. Indicate the test corresponding to the P-values shown.


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: EZH2/H3K27ME3 AND PHOSPHORYLATED EZH2 PREDICT CHEMOTHERAPY RESPONSE AND PROGNOSIS IN OVARIAN CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: no more comments
Experimental design: no more comments
Validity of the findings: no more comments
Additional comments: The authors have addressed my concerns and made significant improvements 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: EZH2/H3K27ME3 AND PHOSPHORYLATED EZH2 PREDICT CHEMOTHERAPY RESPONSE AND PROGNOSIS IN OVARIAN CANCER
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript needs revision for English correctness.
Experimental design: No comment
Validity of the findings: Most comments of the first review were addressed satisfactorily by the authors. There is however a concern regarding the validation of immunohistochemistry for the different proteins measured here. This is important because the study is based mainly on the accuracy of these determinations. The use of PBS as a negative control to demonstrate antibody specificity is not appropriate; acceptable substitutes of primary antibody that can be used for this purpose are IgG or neutralization of primary antibody with immunizing peptide. The authors should run these tests for all antibodies used and include the results as supplementary information.
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: DIRECT AORTIC ROUTE VERSUS TRANSAXILLARY ROUTE FOR TRANSCATHETER AORTIC VALVE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please see below
Experimental design: Please see below
Validity of the findings: Please see below
Additional comments: In the manuscript by Lee et al, entitled “Direct aortic versus transaxillary transcatheter aortic valve
replacement: a systematic review and meta-analysis”, the authors searched for studies reporting clinical outcomes after TAVI performed by direct aortic or transaxillary and analyzed 25 studies. Overall, mortality was similar, but post-procedural complications were higher with the transaxillary approach.

The registration on PROSPERO, the inclusion of studies with VARC definition and the exclusion of very small studies are important elements of this study.
Of course the study has important limitations, mainly the observational design of studies and the low number of patients nd events included.

The search was performed until July 2017. To make this study more interesting and appealing for the contemporary practice, please update it to Oct 2019. Few more studies will be added but will provide more power and more contemporary data.

Medtronic Corevalve and Edwards valves are the most used and for which we have the vast majority of strong evidence, but the authors should clarify the reason why they excluded studies with new valves. How many of such studies were excluded? It would be useful for readers to report a supplementary table with all excluded studies with the reasons and the reference.

Please report in the text or supplement the exact string of keywords used to do the search in each database.

Please add in the abstract the total number of patients analyzed in the 25 studies included.

Abstract conclusion: “both” is not necessary.

Please clarify in all the text including the abstract that the meta-analysis included studies not directly comparing the 2 approaches and reported and pooled rates of events from these studies.

Introduction: the authors state that transfemoral is the preferred approach but in some cases it is not possible. This is true, but the authors should clearly state for readers the reason for this preference. Indeed, transfemoral access is the one that allowed TAVI to be superior to SAVR (Gargiulo G et al, Ann Int Med 2016). In this meta-analysis, TAVI was non-inferior to SAVR and for the first time even in intermediate risk patients, but most of all, when TAVI was transfemorl it was superior in terms of mortality to SAVR. These data were then confirmed by subsequent analyses and trials. Please add this important point and citation.

Figure quality should be implemented.

A mother tongue revision of English language would be needed.


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

Input:
Title: DIRECT AORTIC ROUTE VERSUS TRANSAXILLARY ROUTE FOR TRANSCATHETER AORTIC VALVE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The references used are dated and mostly of only historical significance.  The current manuscript is only about high-risk an inoperable patients.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The manuscript is well written and the methodology sound.  The meta-analysis is dated and fails to reflect current practice in the US where TAVR is approved for all levels of risk.  Discussing STS PROM <8 vs >8 is useless today.


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: DIRECT AORTIC ROUTE VERSUS TRANSAXILLARY ROUTE FOR TRANSCATHETER AORTIC VALVE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 1
Reviewer: 3

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: Specific comments follow:

Summary:  The authors started off with over 400 studies devoted to TAx TAVR and DAo TAVR.  They used certain rules to select studies for a working poll.  The final group consisted of 25 studies.  From these studies, the authors looked at TAx TAVR and DAo TAVR outcomes.  After considering different factors, the authors concluded that DAo TAVR patients had a better outcome than TAx TAVR patients.

Points:
1.	The selected pool of studies is too small and is too selective.  The selection scheme culled out over 90% of the original studies.  The authors eliminated some records, because the study writers did not use precise definitions.  The authors should have attempted to “translate” those studies into common terms.  This would have enlarged the pool of useful studies.  Thus the analysis would have been more useful.

2.	The authors need to review their writing.  The following are major issues:
a.	In APA style, the punctuations follow citations.  The authors had the order reversed.
b.	In APA style, when there are three researchers on a paper, all three are listed.  The authors did not follow this guidance for the one research paper that had three researchers.
c.	In APA style, when the names of the researchers are made part of the sentence, then only the year is placed inside the citation parentheses.  In one spot, the authors did not follow the APA guidance.
d.	A voice should be selected and maintained.  The authors switched back and forth from the first-person pronouns to the third-person pronouns.
e.	Formal writing avoids anthropomorphizing usage.  The authors used personal pronouns to refer to objects.
f.	Parallel structure and a consistent pattern will help the reader to follow the discussion.  The author did not do this with TAx TAVR and DAo TAVR.  And the authors did not do this for sentences that contain a series.
g.	Shorten forms should never be used before the long form.  In several places the authors used the shorten form before using the long form.
h.	Use the active voice.  The authors had active voice, but in a few spots the authors used the passive voice.
i.	Use strong verbs.  The authors used weak verbs in a few places.
j.	Avoid using long sentences with several subordinate clauses.  The authors used very long sentences with several subordinate clauses.


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: DIRECT AORTIC ROUTE VERSUS TRANSAXILLARY ROUTE FOR TRANSCATHETER AORTIC VALVE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: ok
Experimental design: ok
Validity of the findings: ok
Additional comments: The authors did a good job in addressing all comments raised by both reviewers. The manuscript is updated and has improved significantly.


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: DIRECT AORTIC ROUTE VERSUS TRANSAXILLARY ROUTE FOR TRANSCATHETER AORTIC VALVE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: New Review

I am using the authors’ feedback to provide my comments.

See below
Experimental design: See below
Validity of the findings: See below.
Additional comments: I see that the authors revisited this area.  Instead of beginning with 400-plus studies, they started off with 703 studies.  After culling, they came up with about 32 useful studies.  (I ran the math twice and came up with 32 both times.  Double check the list of culled out records, because it appears that one is missing.)
== When I saw the word “finally” in line 149, I thought the authors were adding another 31 studies.  It was not until I was in the “Quality Assessment” section that I realized that the sentence in line 149 was referring to the final group.  I recommend using a different lead-in phrase such as:

	Thus these 31 studies were used in the quantitative meta-analysis…..

I was impressed that these 31 studies looked at 5,055 patient outcomes.  So the final pool of studies was a decent representation.

== In line 80, you stated that you looked at “all relevant studies published before July 2017.”  In Table 1, there are eight studies that were published after 2017.  You might wish to revise that sentence in line 80.

== From a quick read of the submission, I was able to follow the flow of their presentation.  I was not tripped up by the writing.  This is a vast improvement.