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: GLOBAL TRANSCRIPTOME ANALYSIS OF DIFFERENT STAGES OF PREIMPLANTATION EMBRYO DEVELOPMENT IN RIVER BUFFALO
Review round: 1
Reviewer: 1

Output:
Basic reporting: The “Global transcriptome analysis of different stages of preimplantation embryo development in river buffalo (#37890)” is an interesting manuscript with a great amount of data for the early embryonic stages of the river buffalo. The introduction is concise and clear which integrates the reader into the question of the work. However, there are some relevant subjects I would like to address:
-The English language should be improved to facilitate the comprehension of text by an international reader. There are some professionals or companies that offer the language revision. The language could be improved for example, in the text lines: 25-26, 31-32, 53-54, 58-59, 155-158, 170, 173-174, 201, 206-207. Line 123 and 143 “three biological repeats” where “replicates” is a more appropriate term.
- The references should be reviewed. Line 54-55, for example is wrong. The text mention that “Global transcriptome profiling had revealed that gene expression of preimplantation embryo is essential for the embryonic and foetal development (Almen et al., 2009).” Almen et al 2009 refers to human membrane proteome. It does not mention embryo development or any stage of early development. Moreover, at Line 212-214 the authors start the discussion with a wrong information: “Starting from fertilized egg, the embryo undergoes a series of cell divisions to generate blastocyst which includes three distinct cell lineages—trophectoderm, inner cell mass and epiblast cells (Rossant and Tam, 2009).” The epiblast cells are derived from the inner cell mass. The third cell lineage is the hypoblast or primitive endoderm. The original information from Rossant and Tam, 2009 is: “The mouse blastocyst, immediately before implantation, consists of three distinct cell groups: the trophectoderm (TE); the epiblast, which is derived from the earlier inner cell mass (ICM); and the primitive endoderm”. Another example is in the discussion, line 255-257: “Its expression (GRB7) was along with the preimplantation embryo development and the down-regulation at BL stage indicated the EGFR didn’t keep as high level as in 8C and MS stage (Herrick et al., 2017).” Herrick et al 2017 does not refer to embryonic gene expression. In this paper the authors tested the influence of external growth factors in murine embryos cultured individually; “in a sequential medium system containing a defined protein source, blastocyst formation, hatching, and blastocyst cell number and allocation were not affected.” The authors could be more careful with their assumptions and the related literature. In this example they could focus more on GRB7 role itself, and not make indirect statements of its function.
- Regarding the references, along the manuscript the authors should use more recent papers from the literature. One example is in the discussion section, line 235, the description of POU5F1 function has only one reference from 1998. POU5F1 is an important transcription factor and has many papers related to its function. The authors could use more recent work as Daigneault et al 2018 (Scientific Reportsvolume 8, Article number: 7753) where they demonstrate that the “Disruption of the bovine POU5F1 locus prevented blastocyst formation and was associated with embryonic arrest at the morula stage”. This paper corroborates the data found in the present work and could enrich the discussion.
- In line 34, 124-125, 219, and 270, the authors say that 81.94Gb raw reads were generated and they show table1 to support this evidence. Table 1 does not have a complete subtitle and it makes difficult to understand this data, because if the total raw reads are given as megabases (M), the number of total raw reads should be 0.81Gb and not 81,94Gb. The same was told for the clean data that correspond to 75.72Gb and it can be 0.7572Gb.
-The authors could organize the figure2 in a more didactic manner. For example, all clusters with genes up-regulated would be together (e.g 1, 2 and 3), followed by the clusters with genes down regulated (4, 5, 6…), and finally add the clusters with variations of gene expression along development. In line 154 the authors say, “cluster 1 and 4 were genes which were up-regulated”. It is not easy to follow in the figure. If the clusters would be together, like clusters 1 and 2, it would be easier to read the text and search in the figure. Another example is “while cluster 6 and 8 showed down-regulated genes in 2C and BL”. These are two different clusters that show different profiles and could be separated in the explanation to give more details and explore other clusters. If they are talking about down-regulated genes at stage 2C, for example, they could have mentioned clusters 3, 7 and 11 as well.
- The Figure 3 should be described in more details in the text. The authors say that “All of them (genes) conditioned high expression in the preimplantation period and increased the expression (except SOX2) at the BL stage (Fig. 3a, b)”. Figure3 has two graphics, but they chose to talk only about the blastocyst stage. Moreover, the authors did not describe YAP1 expression, but talk briefly about it in the discussion (line245). A similar situation happened in the text describing figure 5b. The authors skipped the explanation for ATF7IP2 in the main text and in the discussion, although it has shown an interesting expression profile. I suggest the authors to comment more about their results in the main text related to Figures 3 and 5b.
- The “Gene co-expression network analysis” (line 166-177) findings is not discussed in the manuscript and the figures from this topic should be labelled, as Figure S2a and b. Table S2 refers to “5 Co-expression modules and genes in each modules”, but Figure S2a has 6modules containing two green modules. Labelling figures S2a and b might help the understanding of the results.
- The “Differentially expressed genes (DEGs) detection” topic is not discussed in the manuscript. It is only mentioned in the Discussion. This topic brings so much information and it deserves to be described in more details to be discussed later. Figures S3 and S4 have low resolution and it should be improved to facilitate the reading. The authors say that (line 193-195) “KEGG pathway annotation showed that the most dramatic variation in the six pairs of comparison was metabolic pathway that should be activated in cell division and differentiation (Fig. S4).” However, for 8C vs MS it shows pathways in cancer and transduction that could be described and discussed in the manuscript. This is quite interesting because the embryo starts its polarization during compaction and there are many papers in the literature, although in different mammals, that could help discussing the findings.
- The raw data was not supplied.
This work has generated an impressive amount of data, therefore they could describe and discuss more the information obtained from these results.
Experimental design: The manuscript is in the scope of PeerJ journal and intend to present new data regarding the transcriptome of buffalo preimplantation embryo stages. However the authors could improve the material and methods with details. Three topics (line 73-92): “oocyte collection and maturation culture in vitro”, “preparation of granulosa cells and in vitro fertilization (IVF)” and “zygote culture in vitro” do not mention “table S3 Formula of buffer in this study”. Moreover, the formula of the solutions in the table do not contain the exact concentration of the solutions, making difficult to replicate the experiment. Those topics (line 73-92) could be more detailed and should refer to any previous work if it has been done before.
The methodology could contain more details regarding data analyses, on topic “Data filtering and genome mapping (line 98-104)”.
On the topic “Transcriptome-wide time series expression profiling” (line 148-165), the authors “employed Mfuzz to perform this cluster analysis (line 151-152)” and found 12 clusters. They do not explain in the text all clusters present in Figure2. Clusters 3, 5, 6, 8, 11 and 12 are not included in the description of the results. If the authors do not consider them to be relevant, they could remove from the manuscript.
Validity of the findings: The manuscript present novelties in the gene expression profile of preimplantation embryonic stages of river buffalo with conclusion linked to original question of this work. Although the authors provide great amount of data; it should be more descriptive and discussed as mentioned in the “Basic reporting” and “Experimental design” topics of this review. Therefore, I recommend the article to be considered for publication after answering the revisions.
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: GLOBAL TRANSCRIPTOME ANALYSIS OF DIFFERENT STAGES OF PREIMPLANTATION EMBRYO DEVELOPMENT IN RIVER BUFFALO
Review round: 1
Reviewer: 2

Output:
Basic reporting: The preimplantation studies of embryos provide key insights into the question of when, where, and how cells take on fate separate. In this paper, the authors used RNA-seq to reveal the transcriptome profiles of four stages (2-cell, 8-cell, Morula and Blastocyst) of preimplantation embryo development. And gave an analysis of the gene co-expression network and differentially expressed genes (DEGs) between different stages characterized genes specific expression. Several TFs, POU5F1, TEAD4, CDX4 and GATAs, were also discussed. I believe it is an important work in deep understanding at transcriptional level about the preimplantation development. However, a careful revision is absolutely needed
Experimental design: Some  parameters of WGCNA were not given
Validity of the findings: Discussion section is not like discussion, but like conclusion. So, the authors should rewrite this section.
Additional comments: The preimplantation studies of embryos provide key insights into the question of when, where, and how cells take on fate separate. In this paper, the authors used RNA-seq to reveal the transcriptome profiles of four stages (2-cell, 8-cell, Morula and Blastocyst) of preimplantation embryo development. And gave an analysis of the gene co-expression network and differentially expressed genes (DEGs) between different stages characterized genes specific expression. Several TFs, POU5F1, TEAD4, CDX4 and GATAs, were also discussed. I believe it is an important work in deep understanding at transcriptional level about the preimplantation development. However, a careful revision is absolutely needed as detailed below.

1.	There are extensive reviews regarding transcriptome analysis in detection of embryo development signature, especially the preimplantation period. However, I failed to find sufficient novel insights to this well-established field. Please give the new findings or conclusions in their paper.
2.	As I know, a lot of studies at transcriptional level about the preimplantation development were not given in the introduction. Please add the progress of early embryo from the transcriptome analysis[Vassena R, et al., Development. 2011; 138:3699-3709; Jiang Z, et al., BMC genomics. 2014; 15:756; Zuo Y, et al.. BMC genomics. 2014; 15:1113; Xie D, C et al., Genome research. 2010; 20:804-815; Biase F et al., Genome Res. 2014 24(11):1787-96; Yan L, et al., Nature structural & molecular biology. 2013; 20:1131-1139; Zuo Y, et al., Oncotarget. 2016 7(45):74120-74131; Hu B, et al., Open Biol. 2019 9(6):190054;Graf A et al., Proc Natl Acad Sci U S A. 2014 111(11):4139-44; Long, C, et al., IEEE Access, 2019 7: 7794-7802; Zhong S, et al., Nature, 2018, 555: 524-528; Fan X, et al., Genome Biology 2015,16: 148; Guo F et al., Cell Research, 2017, 27: 967-988, and so on], I believe this will be helpful for better understanding the research status of this area.
3.	“Gene co-expression and differentially expressed genes” section: Please list the parameters of WGCNA: power, minModuleSize, deepSplit, neworkType 5 etc.,
4.	The “Differentially expressed genes (DEGs) detection” section has similar question. The author used DEGseq and PossionDis to test the difference between each two stages, respectively, however the fold change, P value, Q value were not given.Lack of novelty.


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: GLOBAL TRANSCRIPTOME ANALYSIS OF DIFFERENT STAGES OF PREIMPLANTATION EMBRYO DEVELOPMENT IN RIVER BUFFALO
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I recommend the publication of the manuscript: "Global transcriptome analysis of different stages of preimplantation embryo development in river buffalo". The authors made significant changes in the text and answered all the suggestions from the previous revision. They improved the text and figures and added a scheme as a final figure that made the manuscript interesting and easy to follow.


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: GLOBAL TRANSCRIPTOME ANALYSIS OF DIFFERENT STAGES OF PREIMPLANTATION EMBRYO DEVELOPMENT IN RIVER BUFFALO
Review round: 2
Reviewer: 2

Output:
Basic reporting: The English writing of thismanuscript has been imporoved.
Experimental design: no comment
Validity of the findings: This study is fully organized and systematically done
Additional comments: most of the comments have been revised in the new MS, I have no additional comment for this study.


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

Input:
Title: RELATIONSHIP BETWEEN THE DARK TRIAD AND DEPRESSIVE SYMPTOMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The Manuscript entitled “Relationship between the dark triad and depression: The moderating effect of gender” provides interesting findings on the relationships between the subclinical personality traits of the dark triad and Depression symptoms in a quite large sample. However, there are several things which definitely need to be changed before I can approve publication. The most important point is about the statistical analyses and their interpretation, specifically, including gender into the regression models.
Please see my following review below.

Overall, the article is clearly written, mostly easy to follow and interesting to read.
1.	However, the DT traits are sometimes written starting with a capital letter (e.g. in the abstract) and sometimes starting with a lowercase letter. I would suggest to spell the terms as follows throughout the manuscript: “Machiavellianism”, “psychopathy” and “narcissism”. This is in line with the classic terminology / spelling in scientific papers.
2.	Moreover, while I do understand that the first paragraph of the introduction should be seen as a general introduction into the topic, I am still of the opinion that the associations with other traits, emotional deficits, etc. is depicted (too) broadly. It is well known that there are important differences between the three DT traits in the associations with other constructs. Hence, it would be advisable to not generalize such associations across all three DT traits (as it is done so far in paragraph 1).
3.	Additionally, I am of the opinion that the introduction would be easier to read if the DT traits would be explained before referring to their deficits (hence, one might want to think about changing the order of paragraphs 3 and 4 in the introduction).
4.	The sentence in lines 150-151 is not in line with the actual sample used in the present work. Actually, the present sample shows rather low scores in the DT traits. Therefore, it should not be mentioned that people scoring high in DT should be analyzed. The same is true for hypothesis 5.
5.	Lastly, I do not understand, why hypothesis 4 is only related to sub-dimensions of the DT traits.
Experimental design: The present study is a self-report association study, the research question(s) are mostly well defined and also the gap in the previous literature justifying this research is well explained. However, I have several important comments on the methods.
1.	It would be advisable to include information on the consent (written / informed / …) in the manuscript.
2.	Lines 165-168: The information on the sample are not in line with the information from the SPSS file (e.g. in the SPSS file there are 158 instead of 148 males). Please check all numbers throughout the manuscript carefully.
3.	From what is written in the method section right now, it does not get clear whether the Cronbach’s alphas are derived from the cited studies or from the sample of the present study. It would be great if the alphas derived from the present sample would be presented.
4.	The statement “Third, we explored the possible moderating effect of gender on the significant relationships found in the previous step.” (lines 220-221) is actually wrong. With NONE of the analyses implemented, actual moderation effects of gender were investigated. Actually, not even the correlations between males and females were statistically tested for significant differences (e.g. by a Fisher’s z-Test). To explore moderation effects, it would be best to include gender as well as the gender x DT (sub-)dimension interaction terms alongside the other predictors (main effects of DT (sub-)dimensions) into one regression model. If this is not done, no moderation effects are investigated. Therefore, also the interpretations and even the title are not correct.
5.	However, when looking at the data, it becomes obvious that several of the scales under investigation are not normally distributed. This is especially also true for the BDI-II. Hence, parametric testing does not seem to be appropriate for these data. I strongly suggest to use statistic tests (throughout the complete manuscript) which to not rely on the assumption of normality.
6.	Moreover, reporting effect sizes is very good. However, given the different sample sizes of males and females, the authors might want to think about not reporting Cohen’s d but a corrected effect size (e.g. Hedge’s g) in Table 1.
Validity of the findings: The results are interesting. However, next to the comments on the statistical analyses, I also have some comments on the presentation of the results as well as – especially – regarding their interpretation (these comments built upon the comments on the statistical analyses mentioned above).

1.	In Table 1 it appears as if the scores of the total scales of the questionnaires would be calculated by building sum scores; however, subscales were built by calculating mean scores. For reasons of consistency I would strongly suggest to use either sum or mean scores for all (sub-)dimensions.
2.	In Table 1: Females score (descriptively) higher in VAN (NPI) compared to males. Therefore, the t-value should be positive. In line with this, please check the numbers in Table 1 (and also the other tables) carefully for errors (e.g. rounding errors) and correct them when necessary. When working with the SPSS-File provided by the authors, I sometimes receive (slightly) different values.
3.	Table 1: In the note below Table 1, “SD” is explained by “standard error”. However, SD stands for “standard deviation”. Please carefully check which of the two is reported in the table and change the respective parts / descriptions, accordingly.
4.	I would suggest to use the same order of the subscales when presenting methods / results throughout the manuscript (e.g. compare description of questionnaires vs. Table 1 vs. Table 2).
5.	In the text (in the results section) about the specific results, I would suggest to write the complete names of the scales; otherwise it is hard to follow / understand results.
6.	As mentioned previously, to examine (and interpret) moderation effects of gender, this factor MUST be included in the regression models (alongside main effects of the DT (sub-)dimensions and the respective interaction effects).
7.	Lines 269-270: Actually, Hypothesis 1 is not fully supported. Because in the introduction this hypothesis is referring to all DT traits; however, gender differences were not found in each if the (sub-)dimensions / traits.
8.	Lines 269-274: There is much more and newer research about gender differences in these traits. Hence, also newer literature should be cited.
9.	Lines 686-688: Please include references.
10.	Lines 290-292: Please cite more than one reference if you are referring to “studies”.
11.	322-323: Please cite more than one reference if you are referring to “many studies".
12.	If gender (and the respective interaction terms) are not included in the regression model, actually no moderating effects were tested; Just because some correlations reached significance in one (e.g. the female) sample but not the other (e.g. the male) sample, it does not necessarily has to be due to a moderation effect of gender; instead the differences in significance(s) could also be due to the different sample sizes (in line with this, the differences between the correlations do not seem to be very big). Also, descriptively the sizes of the correlations do not seem to be very big. Hence, the complete paragraph on hypothesis 4 in the discussion (and the conclusion) is not really correct (see already comment above). Only if gender alongside the other predictors (as well as the interaction terms etc.) is included in the one regression model, potential moderating effects can be discussed.
Additional comments: I am sure the authors can tackle most of the above mentioned comments. In conclusion and most important, the authors should think about the specific statistical analyses and, hence, also the discussion of their results. Also potential errors in the reported results should be corrected.
Lastly, I have to minor comments:
1.	Why do the author’s mention their work would be an “original Review” in their comment? I guess it should mean “original research”.
2.	Line 253: It is written “VA” but the scale is actually labeled: VAN. Please correct.


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

Input:
Title: RELATIONSHIP BETWEEN THE DARK TRIAD AND DEPRESSIVE SYMPTOMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors investigated the associations between DT traits, depression and gender among a larger sample of participants in early adulthood. The authors observed that specific dimensions of DT traits were associated with symptoms of depression and gender. The authors further underlined that DT traits appeared to be associated with difficulties in emotion regulation.

Abstract: Please add gender ratio.
“…..DT sub-dimensions depending on gender..”; ok, but on which gender specifically?
Introduction: Generally very well written; references are timely, though, the authors might balance the disadvantages of DT traits against the advantages of DT traits; this holds particularly true, as from the view point of evolutionary psychology and psychiatry, DT would have been sorted out over time, which, however, was not the case. Further, give a closer look at (Sabouri, Gerber, Lemola, et al., 2016; Sabouri, Gerber, Sadeghi Bahmani, et al., 2016).
“Depression is one of the most common mental disorders worldwide.”; please add references and, most importantly, report gender differences.
When describing the DSM 5 criteria for major depressive disorders, please add the time frame and clearly distinguish between the diagnosis of major depressive disorders and symptoms of depression. This is particularly important, as from the Method section it turned out that ‘depression’ was solely self-reported and not based on a thorough psychiatric and clinical interview, for instance with the MINI (Sheehan et al., 1998).
“…but relatively little importance has been assigned to the negative emotional consequences that these people suffer.”; see (Sabouri, Gerber, Lemola, et al., 2016)
“….variables such as SYMPTOMS OF depression (Jones & Paulhus, 2017).”
Collectively, the Introduction was particularly well written; the reader gets a comprehensive overview of the state-of-the-art; hypotheses were theory-based and well formulated. Congrats on you.
Methods:
Report, how you did make sure that participants answered correctly to the items. How did you handle missing data? Report, how much time participants needed on average to complete the questionnaires.
SPSS® version 20.0 (IBM Corporation, Armonk NY, USA).
Report the alpha level of significance.
Results:
“…for the ENT…”; I suggest to spelling out all abbreviations; don’t count too much on the reader’s working memory.
Multiple regression analysis (and Table 3); report in more details, if basic statistical requirements to run a multiple regression analysis were met (Durbin-Watson coefficients; multicollinearity; homoscedasticity); Table 3; it appears that all subdimensions statistically significantly contributed to explain the outcome variable; please check once again very carefully, as such results are rather surprising; further, a t-value of 1.86 most probably was not statistically significant.
Discussion: The authors discussed very well their data.
Conclusion: Although it is a matter of taste, I suggest to substantively shorten the Conclusion section.

References
Sabouri, S., Gerber, M., Lemola, S., Becker, S. P., Shamsi, M., Shakouri, Z., . . . Brand, S. (2016). Examining Dark Triad traits in relation to sleep disturbances, anxiety sensitivity and intolerance of uncertainty in young adults. Compr Psychiatry, 68, 103-110. doi:10.1016/j.comppsych.2016.03.012
Sabouri, S., Gerber, M., Sadeghi Bahmani, D., Lemola, S., Clough, P. J., Kalak, N., . . . Brand, S. (2016). Examining Dark Triad traits in relation to mental toughness and physical activity in young adults. Neuropsychiatr Dis Treat, 12, 229-235.
Sheehan, D. V., Lecrubier, Y., Sheehan, K. H., Amorim, P., Janavs, J., Weiller, E., . . . Dunbar, G. C. (1998). The Mini-International Neuropsychiatric Interview (M.I.N.I.): the development and validation of a structured diagnostic psychiatric interview for DSM-IV and ICD-10. J Clin Psychiatry, 59 Suppl 20, 22-33;quiz 34-57.
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: RELATIONSHIP BETWEEN THE DARK TRIAD AND DEPRESSIVE SYMPTOMS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript titled “Relationship between the dark triad and depressive symptoms” by Gómez-Leal et al. improved a lot and the authors did a great job in the revising the manuscript. It reads very well, the introduction, rationale and hypotheses are easy to follow and understand, and the statistical methods are appropriate. The discussion is very interesting.
I only have some minor comments / suggestions on the manuscript (word-file). Afterwards, I approve publication.

Basic Reporting:

Abstract: “The aim of the present study was to analyse the relationship between the DT and depressive symptoms in order to identify those factors most strongly associated with the development of depression in individuals scoring high on DT.“: I think it would be advisable to include some information on the sub-dimensions in this sentence. Because the sub-dimensions have – as far as I understand – not been investigated in this context so far: “The aim of the present study was to analyse the relationship between the DT including sub-dimensions and depressive symptoms in order to identify those factors most strongly associated…”.

Lines 108-109: “However, relatively little importance has been assigned to the negative emotional consequences that these individuals may suffer.”: I guess by writing “these individuals” it is referred to individuals scoring high in the DT? Please clarify.

Lines 201-202: “A score of 0-13 indicates minimal depression, ...”: It would be great if a reference for these cut-off scores would be cited. In the past, I realized that several different rules for the cut-off scores of the BDI-II exist.
Experimental design: Experimental design:

I acknowledge that Cronbach’s alphas are reported for the present sample. However, it seems like these values are only reported for the complete scales of the questionnaires. As the subscales are also of importance for the present study, it would be advisable to also report Cronbach’s alphas for the subscales. At least it might be possible to give some information on the range in which the Cronbach’s alphas of the subscales lie.

Line 243-244: “Descriptive statistics, t-tests, Spearman’s correlations and Fisher's Z-tests were all conducted using the SPSS® version 23.0”: I guess instead of “t-test” it should be Mann-Whitney U test. Is this correct? Additionally, I am pretty sure that it is not possible to perform Fisher’s z-Tests in order to compare two correlations in SPSS. Please clarify.

Lines 255-256: “We included in the model those DT sub-dimensions that were significant in the correlational analysis”: Was the significance checked in the complete sample or the male / female sample? Please clarify.

Additionally, it would be very helpful if some more information on the GAM would be given. There are several settings when implementing a GAM, which can / must be changed according to the data. Also, as the scores assessing the predictors (DT traits and sub-dimensions) have different ranges (depending on the items included to calculate the score, the response scale, …), I am wondering whether it is necessary to standardize these values. Maybe some more information on this topic can be given. At least it would be helpfull to discuss this issue in the discussion section of the manuscript.
Validity of the findings: Validity of the findings:

Caption Table 2: „ Spearman's correlations between sub-dimensions of DT and depression.“: Correlations with the total scores are also presented in Table 2. Therefore, it might be a good idea to think about deleting the „sub“ before the „-dimensions“ or to put it into parentheses.

Lines 284-288: “When specific gender comparisons were made with respect to the correlations using the Fisher's z-test, it was found that men, compared with women, showed a stronger negative relationship between the narcissism sub-dimensions of self-sufficiency (Z = 2.05, p < .05) and vanity (Z = 2.31, p < .05) with depressive symptoms.”: When looking at Table 2, it seems like actually women show stronger negative correlations than men. Please clarify.

Line 293: “…revealed six significant sub-dimensions…”: As far as I can see, it should be five sub-dimensions.

In the data analyses section, it is stated that gender was also included in the GAM. It would be great to clarify i) whether only the main effect of gender was included or also interaction effects with gender, and ii) whether a gender effect turned out to be significant or not.
Additional comments: Line 400: “… included in the regression model.”: I guess it should be “GAM” (instead of regression model). Please clarify.

Please check all statics – and especially information on significance – carefully, once again. For example, when I calculate the mean age of the sample, I receive M=35.757, which would be rounded to 35.76 years (instead of 35.75). Moreover, when running a Mann-Whitney U test, I receive a p-value of .056 for the gender difference in the CA scale of the SRP-III. However, in Table 1 there is an asterisk behind the number, which would indicate significance. The p-value for the gender difference in the T scale of the Mach-IV is actually 0.009767, hence, two asterisks rather than one would be appropriate. The gender differences in the ENT and VAN scales of then NPI show p-values of .008 and .005, which would need to be indicated with two asterisks rather than 1. Of note, these are just some examples.


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

Input:
Title: RELATIONSHIP BETWEEN THE DARK TRIAD AND DEPRESSIVE SYMPTOMS
Review round: 2
Reviewer: 2

Output:
Basic reporting: In my opinion, the authors took their job to revise the ms very seriously. Congrats on you! The authors have dealt with every issue raised by the editorial board and the reviewers in a thorough and comprehensive fashion. Overall, the ms is in a very nice shape, the content is interesting, references are timely, and the text has a good flow.
Experimental design: see above; no further comments
Validity of the findings: see above; no further comments
Additional comments: see above; no further comments


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

Input:
Title: RELATIONSHIP BETWEEN THE DARK TRIAD AND DEPRESSIVE SYMPTOMS
Review round: 3
Reviewer: 1

Output:
Basic reporting: The manuscript "Relationship between the dark triad and depressive symptoms" by Gómez-Leal et al. improved a lot and the authors did a great job in the revising the manuscript, again.
Overall, the manuscript reads very well, the introduction, rationale and hypotheses are easy to follow and understand, and the statistical methods are appropriate. The discussion is very interesting.
I do not have further comments and agree on publication of the manuscript.
Experimental design: -
Validity of the findings: -
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: CHARACTERISTICS OF THE VAGINAL MICROBIOME IN CROSS-BORDER FEMALE SEX WORKERS IN CHINA: A CASE-CONTROL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.Title: Usually reported as a “case-control study”, rather than case controlled
Abstract Background: I think the implication is that understanding differences in the vaginal microbiome is important for STI control among all women; the authors selected sex workers as an extreme group to look for variations that might be associated with STIs. This could be made clearer.
Introduction: Please do not start off with a limitation. The second sentence is much more engaging and the illegal status of sex work can be addressed in the discussion.
Thank you for acknowledging that sex work, especially trans-border sex work, is a difficult social issue and not problematizing the women who engage in sex work.
Line 60: Lactobacillus is a broad genus; some species have been found to be more protective and indeed produce hydrogen peroxide. Others however, in particular L iners, are less associated with vaginal health. Please update your review in lines 59-63.
Line 63: “the mucosal barrier of the vagina was more easily broken” or disrupted
Line 66: Unclear what “except for pathogen infection” means; also unclear what reproduction associated diseases. Are you explaining that dysbiosis can include BV and is associated with negative pregnancy-related outcomes?
Line 72: The number of community state types varies greatly across the literature; the 5 state model is an appropriate choice, but should be indicated as a choice. The lactobacillus discussion here is much more appropriate and perhaps that earlier section can be included here?
Line 81: The critique of the African study seems unnecessary, as you are not addressing that particular question.
Line 90: Vaginal swabs (cut “information”)
Line 99: For those not familiar with the geography, how close are these two study sites?
Line 100: Please capitalized Chinese Center for Disease Control consistently.
Line 148: p-value should be less than 0.05 to be significant
Line 168: Please provide more explanation than clustered in the right or clustered in the left.
Line 194: Unclear what “divided by” refers to?
line 203: Race, geography, and hygiene practices could all explain these differences.
line 205: This is a new and somewhat unexpected hypothesis – understanding how FSW impact the COMMUNITY microbiome goes far beyond the sampling and methodology of this paper.
line 206: This paragraph needs to be edited, both to improve the language (grammar and typos), but to have a clearer purpose. Are you outlining the strengths and limitations of the study? If so, please state this clearly.
line 218: Please be clearer: Neisseria and Chlamydia are known pathogens and it is not surprising they were found more often among FSW; it is unclear that they alter pH. Overall however, this paragraph is good and provides interpretation and meaning to the results.
Line 266: This is an excellent summary of limitations but should be incorporated into the paragraph starting at line 206. The ideas for future research are good.
Figure 1a: what do the colors and shapes indicate?
Figure 1b: also address in the text, what species loaded onto each principal component?
figure 1c: what does the scale -6 to 6 refer to?
Please include somewhere the proportion of each group with Neisseria and Chlamydia; antibiotic use and the other factors described in the limitations should also be included in Table 1.
Please present the CST by groups.
Experimental design: Abstract results: It is unusual to include STIs themselves in the analysis (Neisseria and Chlamydia), especially as the premise is to look for other vaginal microbes that might be associated with acquisition of Neisseria and Chlamydia.
Line 85: As there are no approved treatments to successfully change the vaginal microbiome, the second part of this sentence seems inappropriate
Methods: It is interesting that you describe community state types in the introduction but do not use them to characterize your samples.
Line 186: Community types are used! Please include this in your methods section as well. The results are very interesting and it is valuable that you were able to examine so many lacto species.
Power: How was the sample size determined? How were the control samples selected – was there any matching on age, ethnicity, etc?
Please state a specific hypothesis you are testing, for example – we hypothesized that FSW would have higher alpha and beta diversity relative to NFSW.
Validity of the findings: Are any of the differences in diversity or PCA still present when Neisseria and Chlamydia are removed? If your hypothesis is that differences in microbiome lead to Neisseria and Chlamydia infection, it is inappropriate to include these bacteria in your analysis. It would be tempting to look for associations between microbiome and Neisseria/Chlamydia among the sex worker samples – but your sample size may be too small.
Line 239: Unclear what you are saying. There are clear shifts in CST between your two groups (in the supplemental materials), even if they do not meet statistical significance (give the small numbers, Fishers Exact might be a better test). How are you defining dysbiosis? I don’t see this anywhere in the results.
line 243: Why would hospital patients be more likely to have vaginal dybiosis?
Conclusion: I would rethink what these results mean. What would the benefit of screening actually be?
Figure 3. Unless you also performed quantitative PCR, I’m unclear that log(Abundance) is a meaningful measure.
Additional comments: Thank you for undertaking this work; I believe the data have potential to be very informative. However, I think the analysis must be updated to exclude Neisseria and Chlamydia and then re-assess your results. In addition, please review appropriate methods for comparing relative abundances of specific species. Finally, I am concerned about your sample size.


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: CHARACTERISTICS OF THE VAGINAL MICROBIOME IN CROSS-BORDER FEMALE SEX WORKERS IN CHINA: A CASE-CONTROL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is an interesting article describing differences found in the vaginal microbiota of female sex workers in southwest China.  The article is well presented and has interesting findings, however there are several places where the English is not up to standard and needs to be revised:
1.  Lines 62-63 are very poorly worded and I can't understand the meaning.  Revision is needed.
2.  Lines 81-82 are also poorly worded.
3.  Line 118 "Hisq" should be "HiSeq"
4.  Line 137 "Line" should be "Linear"
5.  Line 175 "this" should be "these"
6.  Lines 183 and 184 "related" should be "relative"
Experimental design: The use of 97% OTUs in the primary analysis of this data would likely obscure several different species of Lactobacillus which have representative sequences that get grouped together into the same OTU.  Use of 99% OTUs would be far more accurate in this type of study, or better yet use of a more advanced method like DADA2 which isolates biologically meaningful sequence variants.
Validity of the findings: The findings of differences between FNSW and FSW are quite stark and particularly the figure panel b showing PC1 vs PC2 is a perfect separation of FSW and FNSW.  This pronounced of a difference leads me to believe (speculate) that the differences the authors are seeing may be mainly driven by the fact that the FSW's are primarily of Vietnamese descent (crossing into the region at the border) whereas they are being compared to the FNSW's who are likely primarily of Chinese descent.  This is something that the authors do a good job of pointing out in the discussion, however it is not highlighted as a limitation in the abstract.  Readers shouldn't have to dig for this important of a distinction.  Further, Table 1 should include a breakdown of the background of the FSW's and FNSW's so that the reader can see how much of an issue this may be.  How many of the FNSW's were of Chinese background vs Vietnamese?  How many of the FSW's were of Vietnamese background vs Chinese?
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: CHARACTERISTICS OF THE VAGINAL MICROBIOME IN CROSS-BORDER FEMALE SEX WORKERS IN CHINA: A CASE-CONTROL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have made good changes throughout that strengthen the article. There are still many places where grammar could be improved and I recommend one more round of edits to polish the language.
Experimental design: The authors addressed the questions raised well. The sensitivity analysis showing that not all the differences between FSW and FNSW were due to Chlamydia and Gonorrhea are important and should be included. Likewise, the analysis between Vietnamese and Chinese patients is a nice addition. The edits to the tables and figures are helpful.
Validity of the findings: The results are interpreted well and the limitations are appropriate.
Additional comments: Thank you for the thoughtful edits.


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: CHARACTERISTICS OF THE VAGINAL MICROBIOME IN CROSS-BORDER FEMALE SEX WORKERS IN CHINA: A CASE-CONTROL STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have adequately addressed my concerns about better presentation of the manuscript via fixing of typos and correcting of english language deficiencies pointed out.
Experimental design: I did not have any issues with the original experimental design and the authors have addressed any concerns raised by the other reviewer adequately.
Validity of the findings: The authors have addressed the limitations that were pointed out and added the appropriate disclaimers to the findings.  Particularly the issue that FNSW were all of Chinese background is a limitation of the experimental design and is now appropriately pointed out and detailed in the demographics.
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: LIFE HISTORY RESPONSE OF ECHINOPS GMELINII TURCZ. TO VARIATION IN THE RAINFALL PATTERN IN A TEMPERATE DESERT
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This manuscript presents results of a well-designed experiment on the effect of variation in rainfall amount and frequency on success of a cold desert annual in an arid sand dune ecosystem of northwestern China. It addresses an important question in desert plant establishment ecology. The key result is interesting and well-supported, namely that frequency of precipitation is as important in determining life history outcomes in this species as total amount, and that intermediate frequency is optimal, especially when total amount is below average. The paper is basically well-written and well-organized, with only very minor problems with English, and it does a good job of placing the study in the context of previous work.

My main issue is with the way the data are presented in graphical form and in the text.  The bar graphs in Figures 2, 3 and 4 show the full factorial interaction between precipitation amount and frequency, and they present a visually compelling story, particularly in Figure 2. However I found the means separations on these graphs to be very difficult to follow. As these are main effects means separations, I think it would be best to include main effects graphs in addition to  the full factorial interaction graphs, or perhaps even better,  to present the main effects in table form with their means separations.  The full interaction graphs could still be presented, possibly with means separations for the interactions presented when they are significant. The text tends to discuss all the interactions as if they were significant, but in many cases they were not; this needs to be considered in the way the results are described.

I could not find any statistical support in the paper for the means separations in Figure 2; this needs to be included in the ANOVA table. The main effects in Figure 2 are obviously significant, but the focus of the presentation in the results is on the interactions, so it is important to know whether the interaction term was significant. Also, Figures 2 and 5 are the only figures that contains error bars, and these are not defined—are they standard errors of the mean? Why are error bars included only on these graphs and not the others?

Another question I have is with data transformations for the ANOVA. Survival and seed germination are clearly binomial variables that are not normally distributed, and the best way to handle these is to use an alternative analysis that takes the binomial distribution into account. A traditional alternative has been to use arcsine-square root as a transformation for binomial data, but this has been shown to be less effective than a generalized linear model approach that can accommodate non-normal distributions. Seed mass and number and capitulum number could also benefit from log or square root transformations, but I am not sure why a log transformation was chosen for root-shoot ratio. Reproductive effort is never formally defined, so it is difficult to know how it should be treated statistically. It also appears to be a proportion; please clarify how it was calculated. Data transformation is unlikely to change any of the major take homes of the analysis, but the details of the means separations could change.

I would also like to see a better explanation of the variance component analysis.  These variance components do not sum to one, so I assume the remaining variance is error variance? Or is that interpretation too simplistic? There is a citation for this method, which is good, but it is likely not a primary reference for this statistical method.  In any case, a reader should not have to consult a primary reference to understand the paper. This index is stated to account for the ‘percentage of variation that is explained as a plastic effect’, but I am not sure what this means. The interpretation of the results depends on this analysis quite a bit, so it would be good to add a few more sentences to the methods to explain what it really means.

A few minor points:

In Figure 3, panel B needs a Y-axis label—it is possible to figure this out from the legend, but at first glance it looks like a drafting error, so why not include it?

Line 130. Talking about stabilizing mobile dunes after their stabilization creates confusion. Is it that the dune stabilization method with the straw checkerboards and revegetation is not totally effective, so that mobile dunes redevelop? This needs rewording.

Line 147. It is not possible to know exactly when a buried seed germinates—please refer to this a seedling emergence, not seed germination.

Line 180. The period of experimental treatment from spring to summer does not correspond to the life cycle of a winter annual, which starts when the seed germinates in fall. This just needs to be reworded.

Line 195-196. If overwinter survival is a key factor in population dynamics for this species as you state here, and as you measured overwinter survival in the prequel to your experiment, you could report the data. I know they are not part of your experimental design, but it would be interesting to know.


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: LIFE HISTORY RESPONSE OF ECHINOPS GMELINII TURCZ. TO VARIATION IN THE RAINFALL PATTERN IN A TEMPERATE DESERT
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1) In the introduction hypothesis and predicted effects of the rainfall manipulation could be more specific.   Currently the main hypothesis is that changing the rainfall regime would affect the plants somehow.  This is a reasonable expectation but rather unspecific.  Given the number of response variables measured it was very likely that at least some would respond to the treatments.  Can the authors be more specific about which variables and in which direction they expected the plants to respond to each treatment?

2) Similarly, in the discussion section the authors should attempt to reach a more clear and concise conclusion. There were a lot of response variables measured, survival, germination, growth, height, root/shoot ratio, capitula number, etc.  It can be easy to get lost in the details.  I would appreciate a stronger attempt to synthesize the results into a few important points.  For instance, I was surprised for at the strength of the response to rainfall frequency—the effect of rainfall frequency was nearly as strong as the effect of rainfall amount (Table 2).

3) There were errors in the raw data shared in the excel sheet.  These prevented me from reproducing the results as shown in the paper.   These need to be corrected so that the analyses can be reproduced:

 -  In sheet 3.  The number of treatment rows in columns A and B, does not match the number of rows of data in the other columns.
 - In sheet 3, row 105, column ‘C’.  This shows an anomalous plant height of 322 (cm).  This does not appear to match the average height shown for this treatment combination in figure 3c.
 - In sheet 3, reference error on Line 104, column ‘E’
 - In sheet 3, reference error on Line 72, column ‘H’

4)  In Table 2, the numerator and denominator degrees of freedom MUST be given whenever reporting the results of an F-test.

5) The method used to analyze the survival data is not explained.
Experimental design: 1) The “Experimental Design” section should be reorganized to more clearly convey the basic and most important aspects of the design.  I would suggest starting that section with a description of the actual design, which now starts at about line 162.  This part is so important that it should be brought up to the beginning of the section. This section should clearly and succinctly explain the number of treatments, treatment levels, number of replicates per treatment (see below), in one paragraph.  Currently important elements of the experimental design are mixed in with other details about the plants, the measurements taken and the environment. This makes it difficult for the reader to quickly grasp the design.
Validity of the findings: 1) As stated above there appear to be some errors in the excel spreadsheet that was distributed with this manuscript those need to be corrected.

2) All analyses are conducted with 2-way ANOVAS with an interaction.  However, some of the response variables are not well suited to an analysis with an ANOVA.  For instance, percent germination is bounded between 0 and 1 (or 0 and 100%),  and capitula number is a positive integer and is generally a pretty small number.  Neither of these variables match a normal distribution.  Consider whether an ANOVA with normal errors is justified for each response variable.   The authors could consider using generalized linear models for some of these measures, such as a binomial model with a logistic link for the percent germination, and a Poisson model with a log-link for capitula number, etc.

3) Likewise, because of plants dying some of the analyses will be unbalanced (that is have different numbers of replicates per treatment level).  This can be problematic and the authors need to discuss whether the results of the ANOVA are robust to this.

4) The authors need to more clearly state the degrees of freedom for the tests as well.  This is missing from Table 2 and it depends on how the replicates are counted. (mentioned above as well).
Additional comments: 1) In general, I applaud the authors effort in using a rigorous experimental design to investigate the impact of precipitation on annual plant population ecology.  I especially appreciate that both rainfall amount and rainfall frequency were manipulated separately in the experiment.    I also appreciate the care taken to measure many aspects of plant performance.

2) This is only a suggestion, but the data here present an opportunity to build a population model for the species.  A population model would be a powerful way to understand the overall effect of precipitation on this species.  See Compagnoni (2014) for a good example of integrating separate vital rates into a simple population model for an annual plant species.  This may be beyond the scope of the current manuscript so take it as only a suggestion.  It could be a good topic for a future paper.

Compagnoni, A. 2014. Warming, soil moisture, and loss of snow increase Bromus tectorum’s population growth rate. Elementa: Science of the Anthropocene 2:000020.  Doi: 10.12952/journal.elementa.000020

Minor points

•	Line 69:  Discussing plasticity of survival is bit odd.  The survival rate is defined at a population level.  Plasticity usually involves variation among individuals within a population.

•	Line 69:  Is fecundity an “early life-history stage”?

•	Line 70 -74:  Sentence is a bit long and hard to follow, consider revising for clarity.

•	Line 51:  Grammar, pluralize or use definite article “plant communities”

•	Line 136:  Grammar, pluralize “seedlings”

•	Line 172: Grammar, need definite article at start of sentence: e.g. “[The] shelter was assembled…”

•	Line 180 – 181:  Information on the number of replicates per treatment is very important.  This should not be confined to a parenthetical phrase and should be described earlier in the experimental design section.   If I understand this correctly there were 25*12 = 300 total replicates (pots?) in the experiment.

•	Line 241:  How was seedling survival data analyzed? There is a p-value here, but I don’t see any mention of survival  in the analysis section.

•	In Figure 3 the x-axis is somewhat hard to read.  It can be hard to tell which bar goes to which precipitation treatment (“Quantity change of rainfall”).  I think that centering the tick marks below the treatment and making them bigger would help.  Also maybe spreading the groups of bars for each rainfall amount apart would help.


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: LIFE HISTORY RESPONSE OF ECHINOPS GMELINII TURCZ. TO VARIATION IN THE RAINFALL PATTERN IN A TEMPERATE DESERT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The basic reporting is mostly good.  I appreciate the authors work to revise the statistical analyses and re-write the methods section.

The whole manuscript is much easier to read and comprehend in this draft.  The figures are improved as well.

There are a few sentences that are ungrammatical and a few errors in the text.  See the PDF with my annotations attached.

I also appreciate the authors sharing the raw data with the reviewers.  This is really valuable.
Experimental design: The experimental design appears to be adequate.  The authors have improved their description of the design.
Validity of the findings: I have several concerns about the validity of the findings:

1)  I was unable to reproduce the F-values, eta^2 values and p-values found in Table 2.  I used the raw data in sheet 2 of the excel spreadsheet provided.  I tried to reproduce the results of the 2-way ANOVA's with R and with SPSS.   Each time I got different F-values and different eta^2 values than are shown in the table.  I will e-mail the editor the SPSS output from my tests if that is helpful for the authors.

Please confirm the results of your 2-way anovas.  If there are extra steps in the data analysis required to reproduce your results, these should be reported in the main manuscript.

2) The F-statistic for the "Biomass" Anova in Table 2 changed in this draft compared to the last draft.  Please confirm the value is correct.

3) The authors report that they "recorded plant mortality every three days" (Line 194).  Was this true for the entire duration of the experiment (from September to July?).   If mortality was checked regularly the authors may be able to use a more sophisticated survival analyses (not Chi^2) to determine treatment effects on survival.

4) The data for the survival analyses were not provided.  It would be good to include these data with the manuscript.

The figures appear to be correct based on the data provided.
Additional comments: Thank you for your work to revise this.  I have added my comments on the text to the attached PDF document.


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: LIFE HISTORY RESPONSE OF ECHINOPS GMELINII TURCZ. TO VARIATION IN THE RAINFALL PATTERN IN A TEMPERATE DESERT
Review round: 3
Reviewer: 1

Output:
Basic reporting: The basic reporting is good.  There are minor problems of English usage and odd phrasing in the manuscript.  I hope the editors at PeerJ can catch some of these.   Overall it is fairly well written.  Please see comments on the pdf for more notes.
Experimental design: Good.
Validity of the findings: I was able to recreate the ANOVA results reported in table 2 with SPSS.

Thank you for replying to my earlier comments and fixing the table with the updated values.

From a statistical perspective, I am somewhat concerned about one issue.   There are five combinations of the two treatments (amount and frequency) that are completely missing due to the death of all the plants in those treatment combinations. The number of treatment combinations that were excluded due to missingness should be explicitly mentioned in the results.  These missing levels of the interaction can cause large issues when using ANOVA with a fully factorial design.

I recommend the authors conduct a simple ANOVA without the interaction effects between precipitation amount and frequency and confirm that the significance of the main effects are unaffected.

If the main effects change substantially this may indicate some problems and it may be safer not to conduct the full factorial ANOVA.  If the results remain the same, then there may not be an issue--although I am somewhat skeptical of the interaction effects nonetheless.
Additional comments: Thank you for considering my earlier 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: POLYPHENOL-RICH TRAPA QUADRISPINOSA PERICARP EXTRACT AMELIORATES HIGH-FAT DIET INDUCED NON-ALCOHOLIC FATTY LIVER DISEASE BY REGULATING LIPID METABOLISM AND INSULIN RESISTANCE IN MICE
Review round: 2
Reviewer: 1

Output:
Basic reporting: There are no concerns on basic reporting and language
Experimental design: As noted in the review, some points such as pharmacokinetics to determine the active component and dose justification is not reported
Validity of the findings: The data partially supports the findings but the deficiency is that w e do not know which component is active.
Also if the animals simply stopped eating the high fat food.
Additional comments: Reviewer Comments.

There is an urgent need for adequate treatment for the growing NAFLD and hepatocellular carcinoma. This manuscript reveal a natural product  extract that could be beneficial.
Simple editing:
All “et al. “ in the manuscript should be in italics.
Please search the entire manuscript and change.
Line 52. Replace restore with treat and prevent
Line 72. in the south of China, as well as in India and south east Asia.
Line 96. Five standard (lower case s)
Line 107: Delete The primary antibodies and replace simply with Antibodies to
Line 115: Change concentrated to concentrating
Line 112: Define TQPE as Trapa quadrispinosa pericarp extract. Also, the name of the plant must always be in italics.
Line 196. Please change dominated to predominant
Line 239. Define HOMA-IR.  Homeostatic Model Assessment of Insulin Resistance.
Line 321. Please change to used in other clinical trials.
Line 332. ..waste, that are usually discarded
Questions that need to be answered:
Line 153. How do we know that the mice continue to eat the high fat diet after beginning treatment? It is possible that they lost their appetite and ate less. Was the amount of food eaten (or left in food tray) measured?
Line 153. How ere these doses selected. Normally one selects does based on blood levels and scaled down from toxic limits
In the introduction the authors should note if the pericarp of TQ is used in China as natural medicine for liver disease.  Is this used what led the authors to identify the active components?
Also, there are other Trapa species (such as T. japonica and others. Is the activity specific to TQ?
Was there any plasma pharmacokinetic analysis on the TQPE conducted? Steady state plasma levels are essential. It will also show which of the components are actually absorbed. Of the several constituents of the TQPE it is possible that in the intestinal tract some of them may be converted and it is also likely that only one or more components are actually absorbed.  What is the oral bioavailability of each component? This is important and needs to be determined prior to publishing this manuscript. The authors may be surprised that only one or more components are needed and also that there may be conversion in the gut.
Line 245. Was there a dose response between the 15 and 30 mg/kg doses tested in each of the parameters?
It would be helpful to have a Figure of the proposed NAFLD signaling pathways. I note the pathway as the last Figure. That Figure also needs clarification. There are several up and down arrows. Which of these are related to the TQPE treatment. It is not clear to me.


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: POLYPHENOL-RICH TRAPA QUADRISPINOSA PERICARP EXTRACT AMELIORATES HIGH-FAT DIET INDUCED NON-ALCOHOLIC FATTY LIVER DISEASE BY REGULATING LIPID METABOLISM AND INSULIN RESISTANCE IN MICE
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors showed reduction of the diet induced NAFLD associated to obesity in ICR mice due to administration of a specific herb extract (TQPE), from the pericarp of an aquatic plant popular in south of China, which fruit is used as folk medicine and functional food. They also suggested that AMPK/SREBP/ACC intracellular pathway in the liver is involved in the detected effects.

The authors performed:
1- HPLC from the TQPE
2- liver histology by HE from the DIO with NAFLD mice;
3- body and liver mass;
4- AST, ALT, TG, TC, HDLc, LDLc, SOD, and MDA in blood samples;
4- WB in liver samples of AMPK, SREBP, and ACC.

The language of the manuscript is clear. The references provide sufficient background to its proposal.
The figures and tables are well done and convincing.
The raw data is shared. However, it should be interesting for readers to know which animal group corresponds to each band in the supplied PVDF membranes.
The hypothesis is clear and the data presented is adequate to confirm it.
Experimental design: The subject of the manuscript is within the scope of the journal.
The hypothesis is clearly defined and the aims of the study are adequately indicated within the research field.
The ethical approval for animal usage is described in the manuscript, however, I think it should include the following information in the Methods section and or in the legend figures and tables:
1- the total number of animals used in each parameter analyzed;
2- the entire period of HFD intake;
3- the via of administration of the TQPE;
4- if the animals were euthanized after any period of fast or in  fed condition.
Validity of the findings: The data presented is convincing. The statistical analysis performed is adequate, however there are 3 minor points that I would like to hear from the authors:
1- Is there any effect of the TQPE in control chow animals, such as reduced insulinemia or body weight?
2- Is there any impact of the TQPE treatment on food and or water intake?
3- Are the elevated AST and ALT blood levels associated to any other hepatic injury histological marker?
Additional comments: The manuscript is very easy to read and the figures are well presented.
The inclusion of the translational information regarding the equivalent dose for human is positive, as well as the economical aspect of identifying the potencial beneficial usage of the pericarp since it is an agriculture wast.
However, I would like to see included in the manuscript some information regarding the above mentioned 7 points.


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: POLYPHENOL-RICH TRAPA QUADRISPINOSA PERICARP EXTRACT AMELIORATES HIGH-FAT DIET INDUCED NON-ALCOHOLIC FATTY LIVER DISEASE BY REGULATING LIPID METABOLISM AND INSULIN RESISTANCE IN MICE
Review round: 3
Reviewer: 1

Output:
Basic reporting: The Authors have provided reasonable responses to questions and have made the edits requested.
However, it is also required that some of the clarifications are added into the text for the readers, such as the determination of bioavailability of the active entity in future studies.
Experimental design: The experimental design is fine except that the reason for choosing the doses need to be added
Validity of the findings: The results presented are valid except that the authors must add that the mice simply decreased the amount of high fat diet they ate because of the treatment they were receiving. The food intake and the treatment dose intake was not measured so some estimate of what was observed that the animals were eating normally should be mentioned.
Additional comments: Same as above
A couple more edits are marked on the attached manuscript. It will be helpful if marked (Tracked edits) are returned so I do not have to red the whole paper again!


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

Input:
Title: POLYPHENOL-RICH TRAPA QUADRISPINOSA PERICARP EXTRACT AMELIORATES HIGH-FAT DIET INDUCED NON-ALCOHOLIC FATTY LIVER DISEASE BY REGULATING LIPID METABOLISM AND INSULIN RESISTANCE IN MICE
Review round: 3
Reviewer: 2

Output:
Basic reporting: The revised version of this manuscript is improved. However, there are some minor points I would like to be considered:

1- line 155: instead of "... sacrificed..." I suggest ... euthanized...

2- Please include how long the mice were fasted before euthanasia.

3- line 241: instead of "... insulin signal." I suggest to include ... insulin blood level and action.

4- line 257: instead of "... repaired prohibition of..."I suggest  ... kept the similar level of IRS-1 and AKT phosphorylation than the detected in the control mice.
Experimental design: Please, include the number of hours the mice were fasted before euthanasia.
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: INVASIONS OF GLADIOLUS RUST IN NORTH AMERICA ARE CAUSED BY A WIDELY-DISTRIBUTED CLONE OF UROMYCESTRANSVERSALIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors developed SSR markers to look at allelic and genotypic diversity in 92 isolates of Gladiolus rust. Genome sequence from three US isolates was used to design SSR markers. Markers were first tested on the three US isolates before checking them on seven additional isolates from US. Eight markers that performed well in the initial PCR screen were then multiplexed in two sets of four and run on all 92 isolates. The manuscript is well written and presented logically. However, I have few concerns that I list below.

Major concerns
----- --------
1. This is an obligate biotroph. How was it determined that the assembled regions belong to the rust fungus? I understand that the authors detailed in the methods section how the spores were collected and allowed to germinate on an antibiotic solution. But it would add to the manuscript if the authors check that the contigs selected for marker development are of fungal origin. There are several tools available to sort bacterial and fungal sequences or a simple BLAST might work.

2. How do you explain the negative results in Table S1 for Aus and NZ? It looks like that the negative results were ignored.
Not all Aus/NZ isolates group with US/MX/CR isolates. Three (Ut1841, Ut2048, Ut497) out of four markers in the second multiplex did not perform well. Even if for one moment the results from multiplex 2 are excluded and only look at results from markers in multiplex 1, only 3 and 4 isolates from NZ and AUS, respectively, have results from all four markers.  What could be the reason?
Sample size? There are 5 isolates from Costa Rica but these all have similar genotypes.
Age of the sample? The NZ isolate NZ71696 was sampled in year 2000 performed better than other NZ samples collected in 2004 or 2006.
So, then how do you explain the lack of results for Aus and NZ, for markers that work well for isolates from US/MX/CR?

3. If the lack of results for markers in NZ/Aus isolates is biologically relevant, then clonality still exists but is limited to isolates from North America, US/MX/CR.
Here is one way to look at the data. In Table S1, first exclude results from poorly performing marker Ut497. Then, check if results from at least five out of the remaining seven markers are consistent, we obtain 9 US, 5 CR and 57 MX isolates, but only 2 NZ (NZ99990, NZ71696) and 4 Aus (VPRI 21344, VPRI 21238, VPRI 20858, VPRI 20881) isolates. Based on Figure 1, introductions in Aus/NZ preceded US/MX/CR. Could these isolates in US/MX/CR be derived from Aus/NZ?

If so, then the title could be modified to "Invasions of gladiolus rust in North America are caused by a widely-distributed clone of Uromyces transversalis"

4. Although Ut497 was a poor performing marker, it surprised me that it did not work on three US isolates, including two of the sequenced isolates.

5. Explain how scoring is done for the multiplex reactions either in Results or Methods section, and what does 'a small peak below the threshold peak size of 500' mean.

6. What is the implication of a NOPEAK? What does NOPEAK mean? Does it mean that there was a PCR product visible on the gel but no detection during fragment analysis, or, no product could be visualized on the gel.

7. Table 4: Why these six markers? This analysis can be done for all the 25, even the original 60, markers, as the genomes sequences are available for the three isolates.

Minor concerns
----- --------
1. In the whole genome sequencing, both the dikaryons have been sequenced. Reads can be mapped back to the assembly to find SNPs in regions of interest.

2. It would help to list a few examples where allelic and genotypic diversity based on SSRs was in agreement with results obtained from other types of markers such as or SNPs derived from GBS or whole genomes.

3. Lines 259-260: Was the number of shared contigs higher in the two FL11 isolates versus the CA11?

4. Line 279: What criteria was used to select these eight markers? It is explained in the Methods but not in results. Also please expand on the multiplexing scheme in the results.

5. Being a dikaryon, is it a surprise that most markers were polymorphic?

6. Line 349: What is the basis of this calculation? Please elaborate.

6. Table 1: Make the state/Country as a separate column

7. Table 2: Include the number of markers in the heading

8. Table 3: Please add another column that lists the total reads per isolate. Then change the column headings to reflect Filtered for quality, and Reads that assembled.

9. Table S1: It should be cited in the 'Genotypic analyses' section in Results.
    Add a footnote on what these numbers means. Also, add a footnote on how is 'NA' different from 'NOPEAKS'.

267: Where is the term 'electropherogram' defined?
283: Isolate(s) from South Africa not listed in Table 1
289: Where has been the 'scoring threshold peak size 500' been defined?
296: What was the rationale for choosing a monomorphic marker?

TYPOs
242: Remove the double space - "  reads  " to " reads "
243: 'insert size', not 'insertion size'
244: Expand PF
245: Either list three numbers or remove 'respectively'
248: Add percentages along with the number of reads that were assembled
249: Remove '(contigs)'
259: Delete 'microsatellite containing'
264: Add 'sequenced', as in three sequenced isolates


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: INVASIONS OF GLADIOLUS RUST IN NORTH AMERICA ARE CAUSED BY A WIDELY-DISTRIBUTED CLONE OF UROMYCESTRANSVERSALIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: DeLong et al. provide a clear manuscript about the efforts that led to the identification of high degrees of clonailty on an important Urediniomycete affecting Gladiolus species.

The document is clear, professional and very well written. Each of the sections is well documented and include all necesary information for a good report. The authors do a great job at providing sufficient background information to guide the reader throughout the introduction while also stating the research objective clearly.

While the evidence presented is clear and objective, I feel that the only shortcoming is the absence of a main figure that can summarize the results in a simple and clear manner. In my opinion, a revised version of Supplementary Figure 1 would suffice to present a summary of the results and the main findings.
Experimental design: The experimental design presented by DeLong et al. is solid and adequate for the research question posed. The knowledge gap is clearly stated and the methods used in the current manuscript aim to fill this gap.

Minor comments on stating parameters and tools for repeatability have been added to a revised version to the manuscript, but none of these comments convey a major concern for the experimental procedures used in this research.
Validity of the findings: The findings of this manuscript present a puzzling but common feature of fungal plant pathogenic organisms: A link between clonal reproduction and large scale movement of these disease causing organisms.

The results found here are very relevant to the study of the evolution of fungal organisms, and provide more evidence on the uniqueness of the evolutionary patterns of fungal and fungal-like organisms. All the results are well supported and solve the proposed research questions by the authors.

While the current era of genomics seems to be moving fast into whole genome data genotyping techniques, reports like this are important as they remind researchers that traditional markers are still useful and necessary in plant pathology or other sciences that require markers that can be screened rapidly and are cost efficient.

There are some minor comments attached on the presentation of some of the discussion/conclusions, mostly on the extrapolation of the results from microsatellite data with the biology and pathogencity of the organism. These comments should be addressed so the authors can also provide a critical view of their findings as well as future perspectives of what can be done to evaluate if the lack of genetic diversity found in microsatellite data truly reveals a general lack of divergence in this species, or if other techniques can provide a more accurate view of the evolution of these organisms.

These comments are minor and don't compromise the structure of the manuscript nor the main findings, but are necessary for a more objective view of some of the findings presented here.
Additional comments: Overall, the manuscript presents a very interesting finding on this fungal species. The manuscript is very well written and easy to understand; the methods are solid and the results support the hypothesis proposed.

A few minor comments have been added to the manuscript. Mainly, the absence of a results-driven figure makes gives the manuscript a feeling of incompleteness. However, Sup. Fig. 1 can be slightly improved and presented as evidence for the main findings of this report. Subsequently, some of the speculation of the lack of polymorphisms in SSR v.s. clonality of the species need to be either better justified or be presented with more carefulness. Other minor comments are added to the manuscript directly.

None of these minor comments affect the overall findings. Great manuscript overall.


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: LONG NONCODING RNA MEG3 SUPPRESSES PODOCYTE INJURY IN DIABETIC NEPHROPATHY BY INACTIVATING WNT/Β-CATENIN SIGNALING
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.overall writing is poor and needs significant improvements
2. Literature review- The authors did not have enough background literature search and I was bit lost as to why they choose to study MEG3 Lnc out of nowhere. They need to do more literature search and make it clear in the introduction as to why they choose MEG3. It was kind of choppy, they explained MALATI Lnc and jumped to MEG3
3. article structure-  They have done good job of data presentation
Experimental design: 1. The study conducted is original and fits the  Aims and Scope of journal
2. They need to emphasize Why the whole project was undertaken, research question is poorly defined.
3. The research performed is rigorous, couple of issues need to be addressed as explained in comments to the author
4. Materials and Methods need to be more specific there were instances where I was lost as to how did they do the experiment. A. cell counting protocol- they do not mention how they differentiated live and dead cells.  B. ROS production-I am confused what kit they used.
Validity of the findings: 1.The study is well designed, data is presented well and the reader understands to logical flow
2. The study need to be more robust. The authors assume that when plasmid is injected i it is taken up only by podocytes- it can be taken up by any tissue  and observed changes could be result of changes in general metabolism.
3. Conclusions are well stated and data supports cause and effect in-vitro. However in-vivo data needs more rigorous testing.
Additional comments: Major comments.
1. writing needs significant improvement. The reader is often confused and lost because of the poor language usage. Logical connection, cohesion  and  flow is lacking. Please do extensive literature review and rewrite the introduction as to why LnC MEG3 study was undertaken in DN.
2. Materials and Methods section writing needs significant improvement- please write in brief the principles involved for all the methods. For example ROS was detected by ELISA- I am confused they detected ROS or ROS modified proteins?
3. In Figure 1C- DMSO was used  - please explain why this was used
4.They used pCDNA 3 vector- This could be taken up by any tissue in the body and all other cell types in the kidney. What makes the authors believe that effects observed are specifically because of podocytes in the kidney . could you please give more evidence to show MEG3 is expressed only in podocytes ?.
5. The authors should also carry out blood glucose and other parameters after administering MEG3  to rule out MEG3 Lnc altering the metabolism.
6.  In Diabetic Neprhopathy  almost all cell types are affected.  can the authors  give more evidence to rule out changes in other cell types in the kidney after injecting MEG3 Lnc?


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: LONG NONCODING RNA MEG3 SUPPRESSES PODOCYTE INJURY IN DIABETIC NEPHROPATHY BY INACTIVATING WNT/Β-CATENIN SIGNALING
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comments.
Experimental design: In the current study, Xiajing Che and colleagues have explored the role of Long noncoding RNA MEG3 in diabetic nephropathy. The methods have not been sufficiently described. In particular, how the in vivo studies were carried out specifically in Figure-3 has not been described. How was MEG3 overexpressed in podocytes in vivo using a plasmid?
Validity of the findings: Although the in vitro studies with high glucose treatment and expression studies in patients and animal tissues are novel. The functional studies have not been described well. The studies described in Figure-3 are particularly troublesome. How were the authors able to overexpress MEG3 in podocytes in vivo using a plasmid? This method is not described in the methods sections. To be able to overexpress a gene in vivo using pcDNA3 plasmid is currently technically impossible. These results raise serious questions on how the experiments were carried out and the data interpretation.
Additional comments: The data presented in Figure-3 raises critical questions on the validity of this study.


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: LONG NONCODING RNA MEG3 SUPPRESSES PODOCYTE INJURY IN DIABETIC NEPHROPATHY BY INACTIVATING WNT/Β-CATENIN SIGNALING
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Please include  luciferase reporter map
double check ROS detection method- ELISA kit- I am confused what did you detcet?


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: LONG NONCODING RNA MEG3 SUPPRESSES PODOCYTE INJURY IN DIABETIC NEPHROPATHY BY INACTIVATING WNT/Β-CATENIN SIGNALING
Review round: 2
Reviewer: 2

Output:
Basic reporting: No Comment
Experimental design: No Comment
Validity of the findings: No Comment
Additional comments: The authors have addressed all the concerns/comments adequately.


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: MULTIPLE ECOSYSTEM SERVICES FROM FIELD MARGIN VEGETATION FOR ECOLOGICAL SUSTAINABILITY IN AGRICULTURE: SCIENTIFIC EVIDENCE AND KNOWLEDGE GAPS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The main problem throughout the paper is of repetition of information, lack of logical flow especially in the Results and Discussion Section, as well as long (sometimes ambiguous) sentences that make the reading difficult. Also, the writers kept switching between past and present tenses which makes the paper hard to read. Moreover, some facts appear to be contradictory because of poor writing.
Experimental design: The methodology used is adequate and can be replicated but there is a problem with the organization of facts. Although related facts are organized into paragraphs, they are sometimes unnecessarily long with a poor logical flow.
Validity of the findings: Although the paper presents an objective (given in the first few lines of the Methodology Section) with a worldwide view, the conclusions (and scarcely few future directions) are mainly directed toward Africa. Thus, the scope of the paper is broad at the beginning but narrows down to Africa, particularly Tanzania. I think there is not sufficient justification that the recommendations are applicable to "other tropical countries" as presented in the paper.
Additional comments: Though this review paper contains a lot of valuable information and has a broader view (in respect of groups of organisms discussed) compared with previous ones like Vickery et al. (2009), it lacks the elements that will sustain the reader's interest in reading the paper. Because review papers are generally long, readers become bored if they have to read sentences over and over the get the meaning. The paper has many interesting facts but I had to read most sentences two or three times to get the meaning and how they relate to other preceding sentences.


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: MULTIPLE ECOSYSTEM SERVICES FROM FIELD MARGIN VEGETATION FOR ECOLOGICAL SUSTAINABILITY IN AGRICULTURE: SCIENTIFIC EVIDENCE AND KNOWLEDGE GAPS
Review round: 1
Reviewer: 2

Output:
Basic reporting: There are some comments in the text in order to improve clarity.
Experimental design: The fact the Africa has little publication/work on the issue should be pointed out in a better way, there are other regions with same gap.
Validity of the findings: All report all well referenced.
Additional comments: Clarify what are the meening of WEEDS and how they differ from NON-CROPs.

Table 1 is not necessary, it can be convert it in text. Figure 2 is already in the text, thus it is not necessary.

Figura 4 should be improved.


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: MULTIPLE ECOSYSTEM SERVICES FROM FIELD MARGIN VEGETATION FOR ECOLOGICAL SUSTAINABILITY IN AGRICULTURE: SCIENTIFIC EVIDENCE AND KNOWLEDGE GAPS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Generally good; however, a few sentences have to be moved to different sections to improve the logical flow
Experimental design: No comment
Validity of the findings: No comment
Additional comments: See attached 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 DISCRIMINATIVE POWER OF THE REPROQ: A CLIENT EXPERIENCE QUESTIONNAIRE IN MATERNITY CARE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper needs an edit since few word are either missing or needs replacement like line 47- "mortality" repeated, line 57-rewrite sentence.
I suggest adding few lines about relevant prior literature.
Add for reference line 108: Previous results indicate that ratings of the 1st half of pregnancy remain highly associated (ICC=0.80) with ....."
For line 112-121- I suggest to add details in a tabular manner for better understanding.
Line 129-132- mention duration of data collection
Experimental design: Study design is well explained.
No comments
Validity of the findings: Study findings are explained in detail and adequate for the research topic. Writing column variable name rather than writing column number will enhance understanding of the readers for example line 236-237.
As per Table 1: 6% antenatal women and 8% postpartum women participated from low education level. The possible impact of low education on study findings can be discussed in ‘discussion section’.
Additional comments: No additional comments


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

Input:
Title: THE DISCRIMINATIVE POWER OF THE REPROQ: A CLIENT EXPERIENCE QUESTIONNAIRE IN MATERNITY CARE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Statistically, this has been very well approached and vividly documented
However, while reading it gives a feeling there is more of statistical focus.
Adding quality improvement and challenges in assessing the quality in reproductive care, what has been conventionally followed, what are the lacunae of those approaches might add value to this article
Experimental design: Since we expect women who experienced complications may experience more of negative picture and complications are commonly occur during childbirth and postpartum attempt for sensitivity analysis will ensure the robustness of the findings.
In the multi-level nested/hierarchy there is client, provider (if one hospital is managed by multiple providers), hospital and perinatal unit. Authors have explained the reason for not including the provider in the model.
Is there any reason not to include hospital as one of the levels? (unless the hospital and provider are equivalent).
Though Appendix B clearly describes how they decided to choose between random intercept and random slope model bringing that small paragraph to the main manuscript will increase clarity of approach.
Validity of the findings: In the result section, authors have mentioned the response rate as 32%. But in the method, it was mentioned everyone participated and gave informed consent. This might need a clarification
If the response rate was 32% authors can’t claim for generalizability as mentioned in the discussion section unless they could ensure the responders and non-responders are similar in terms of age, education and self-rated health.
Additional comments: Statistically, this has been very well approached and vividly documented
However, while reading it gives a feeling there is more of statistical focus.
Adding quality improvement and challenges in assessing the quality in reproductive care, what has been conventionally followed, what are the lacunae of those approaches might add value to this article
In the result, section authors have mentioned the response rate as 32%. But in the method, it was mentioned everyone participated and gave informed consent. This might need a clarification
If the response rate was 32% authors can’t claim for generalizability as mentioned in the discussion section unless they could ensure the responders and non-responders are similar in terms of age, education and self-rated health.
Since we expect women who experienced complications may experience more of negative picture and complications are commonly occur during childbirth and postpartum attempt for sensitivity analysis will ensure the robustness of the findings.
In the multi-level nested/hierarchy there is client, provider (if one hospital is managed by multiple providers), hospital and perinatal unit. Authors have explained the reason for not including the provider in the model.
Is there any reason not to include hospital as one of the levels? (unless the hospital and provider are equivalent).
Though Appendix B clearly describes how they decided to choose between random intercept and random slope model bringing that small paragraph to the main manuscript will increase clarity of approach.


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 DISCRIMINATIVE POWER OF THE REPROQ: A CLIENT EXPERIENCE QUESTIONNAIRE IN MATERNITY CARE
Review round: 1
Reviewer: 3

Output:
Basic reporting: My compliments for a good written paper. Nevertheless, could you clarify the following thins:
line 79-80: 'Poor outcomes are infrequent, and specific low performance (for clients and units) into one direction easily averages out into other directions.'
Also, please use another word for 'true' (e.g. line 82). (Parts of) results of statistical analyses should not be associated with 'true'.
line 85: I believe 'should' should be removed from the sentence.
line 131: 'from several ongoing regional observational studies', what does this mean?
line 141: 'Since each hospital can be assigned, uniquely to one perinatal unit'. What about hospitals in urban areas (the 'randstad'? How did you assign these hospitals to perinatal units? Or did you remove those hospitals?
line 382: For the future, we recommend the use of ReproQ in maternity care.' For what purposes, the ReproQ may be used?
Experimental design: Thank you for providing a well defined study and good described design. I have one major comment. In line 335 you state that you avoided overfitting by limiting case-mix correction. However, how did you make sure that your corrected sufficiently for case-mix? Could you please elaborate on that (plus back up with the relevant statistics for your models)?
Validity of the findings: See comment in 2.
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 DISCRIMINATIVE POWER OF THE REPROQ: A CLIENT EXPERIENCE QUESTIONNAIRE IN MATERNITY CARE
Review round: 2
Reviewer: 1

Output:
Basic reporting: satisfactory
Experimental design: The appropriate design has been chosen to answer the research question

The comments given for the previous manuscript is well addressed.
Validity of the findings: The statistical approach and reporting of those methods were clearly documented and similar to the previous section suggested comments are addressed.
Additional comments: We appreciate authors efforts to address all the queries and rebuttal for each point.


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: COMPARATIVE PROTEOMIC ANALYSIS OF PEPPER (CAPSICUM ANNUUM L.) SEEDLINGS UNDER SELENIUM STRESS
Review round: 1
Reviewer: 1

Output:
Basic reporting: it is satisfactory.
Experimental design: it is satisfactory.
Validity of the findings: Real-Time PCR (qPCR) Veriﬁcation is neede.Some differential proteins should be selected to perform the mRNA expression level veriﬁcation in order to validate the labeling results.
Additional comments: No comments.