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: INCREASED MENTAL STRESS AMONG UNDERGRADUATE MEDICAL STUDENTS IN SOUTH-WESTERN SAUDI ARABIA DURING THE COVID-19 PANDEMIC
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an interesting area of research however the literature review has not been carried out in detail as there are a number of studies that have been published on this topic. The introduction does not provide a sufficient critical approach either.
Experimental design: Very limited when considering methodological approach.
Validity of the findings: I am not sure if there are any novel findings since some of the recent papers have already provided a good amount of data from Saudi Arabia. Thus I would like to encourage the team to review current studies and then collect more data and carry out more sophisticated analyses of the data.
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: INCREASED MENTAL STRESS AMONG UNDERGRADUATE MEDICAL STUDENTS IN SOUTH-WESTERN SAUDI ARABIA DURING THE COVID-19 PANDEMIC
Review round: 1
Reviewer: 2

Output:
Basic reporting: First of all, I would like to thank you for the opportunity to review the paper entitled: Increased mental stress among undergraduate medical students in south-western Saudi Arabia during the COVID-19 pandemic. In this research, authors  evaluate the stress among undergraduate medical students during the Covid -19 Pandemic.  In general, the paper has an interesting and relevant topic for researchers and medical professors and the written level of English is very good and easy to read. However, the methodology and main results need to be rewritten. With your permission I recommend the following:
Some information are confusing:
The information about first survey - what are the questions and if also the Cohen questionnaire was completed? We need more information about this two topics.
In the second survey, the number of the students is almost double comparing with first survey? Did you collect data from the same students in different periods or from different students in different moments? If your data are not collected from the same students, how you will control the stress level?
Experimental design: Line 191 - T test was used to compare the P.S.S, there is any table with t values?
It was difficult to understand why did you use a multivariable analysis and after a correlation, there is any arguments for using this methods?
For a better understanding please insert the statistical description for each variable in a table. Please, insert in a table all the results from the questionnaire and after the table results comparing both sessions, including what was the statistical test that you use, the T test or Mann Whitney U test and effect size for each comparison.
Validity of the findings: Your work was impressive and after the researchers from the field, the second actor are the professors. Please follow the needs and describe your work for both size (for researchers insert all the details for replicate the study with other representative samples and for professors - describe more clearly the technique that you used in table 2 and 3.
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: INCREASED MENTAL STRESS AMONG UNDERGRADUATE MEDICAL STUDENTS IN SOUTH-WESTERN SAUDI ARABIA DURING THE COVID-19 PANDEMIC
Review round: 2
Reviewer: 1

Output:
Basic reporting: Congratulations on your work in answering the questions. After rereading the manuscript, there are many more doubts about the presentation of your data, especially the logistic regression and the spearman correlation. Unfortunately, even if the number of participants is large (1074 students), the results are confusing. For logistic regression, you used multinominal logistic regression, if I understood correctly, please provide us with more information about the model. There is an example how to report the results Andy Field book (Discovering statistics using SPSS) page 284-312. I was trying to understand your results, but I'm still confused.
Regarding the table with the correlations Spearmon (rho), the results are very poor, correlation spearman = 0.14, p.001? I think this is a mistake. Thus, please redo the results section, indicating for each variable used, including the stress questionnaire, the description of each variable, and then argue the use of each analysis in order to understand the results. Your work is extraordinary, but the results are not clear.
Experimental design: The experimental design is well describe.
Validity of the findings: A detailed description of the approach is needed. We do not have enough data on each variable. Another relevant aspect concerns the reasoning of each analysis, why these analyzes were used and what the impact is on the results. Please follow the steps in reporting the results.
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: INCREASED MENTAL STRESS AMONG UNDERGRADUATE MEDICAL STUDENTS IN SOUTH-WESTERN SAUDI ARABIA DURING THE COVID-19 PANDEMIC
Review round: 2
Reviewer: 2

Output:
Basic reporting: Overall, I found the paper well written and professionally and appropriately structured. There a few very minor edits required to the language.
Experimental design: This is a cross-sectional and therefore observational study. The aims of the research were clearly stated and appropriately placed at the end of the Introduction. The analyses were appropriate for the study. As the data were provided, I did obtain some slightly different results than the authors. In general the authors provided sufficient information to allow for replication of the results except in those instances referred to in this review, when it is not clear why some slightly different coefficients were obtained in my analysis.  I apologise if I am in error.
Validity of the findings: Overall I support the findings reported by the authors, with the exception of a few issues mentioned in this review.
Additional comments: INTRODUCTION
(with line numbering beginning on P1 of the review pdf)
Line 64: ‘Although young adults are at low risk of COVID-19 infection (Vieira et al. 2020), college students may represent a vulnerable population for mental health problems during the pandemic’.  This may have been the case with the original strain of Covid19, when the study was done, but is not necessarily the case now. I suggest some qualifying statement perhaps ‘Although young adults are at low risk of COVID-19 infection from the original strain of the virus (Vieira et al. 2020)’, or perhaps raise this point in the Discussion.
Line 75 – 77: ‘Therefore,  we hypothesize that COVID-19-related challenges would represent a more considerable stressor and result in a more significant increase in perceived stress during the pandemic’.
I suggest deleting the word ‘more’ from this sentence (both times it occurs), otherwise the reader is left asking ‘more than what’?
Line 95: ‘…exceeds the pre-pandemic prevalence among the same college students’.  This is not entirely the case as the current study did not use the same college students; rather the current study used similar college students.
STUDY PARTICIPANTS AND PROCEDURE
For both the first and second surveys, were the students a mix of years, eg 1st, 2nd, 3rd etc? All we are told is that they are undergraduates.
Line 112: What is meant by ‘level eight’ medical students?  Please clarify what is meant by ‘academic level’.
Line 128: It would be helpful to know the denominators here: 353 out of how many, and 704 out of how many?
Line 134: ‘Students were stratified by academic level with a cluster chosen from each level’. So was the academic level the cluster? Also, the way the sentence is written makes it sound as though there could have been a number of clusters for each level, out of which one cluster was selected. The next sentence says ‘All listed students in each group were involved in the study whenever possible’. Is a group the same as a cluster?
Line 133: Currently reads ‘In the second one…’, suggest changing to ‘In the second survey…’, but once again, please clarify what is meant by ‘level.’
STUDY TOOL
Line 179: I understand word count limitations, but please briefly explain how the eight Covid questions were validated, e.g., how many experts, what process was followed, etc.  Also, Cronbach’s alpha specifically measures internal consistency reliability, so I suggest ‘…and Cronbach’s alpha was 0.86, indicating internal consistency reliability of the scale’ (2 decimal places here are sufficient).
DATA ANALYSIS
Line 194: Please cite SPSS as IBM SPSS Statistics version 22.
Line 195 – 199: Please re-write as ‘Chi square test of independence was used to  compare the personal and academic data of the students before and during the COVID-19 pandemic. A multivariable logistic regression analysis model was used to assess the independent factors of high perceived stress in the whole sample.’
RESULTS
Line 210: Please re-write as follows ‘The two groups were matched on age, sex etc’. Next sentence, beginning ‘However…’, change to ‘…the family income of the two groups was statistically significantly different’.
Line 212: In text here, the income variable is reported as ‘satisfactory’ or ‘unsatisfactory’ but in Table 1 the levels are ‘Sufficient and exceed’, ‘Sufficient’ and ‘Insufficient’. Check Table 2 also for this variable. Please use the same terminology between tables and text. Was the difference between these two levels determined by obtaining the Adjusted Standardised Residuals from the Crosstabs procedure in SPSS?
The Age variable is reported as <=22 and +22.  For clarity and to avoid confusion, I suggest reporting the +22 as => 23.  This also applies to Table 2.  What was the justification for dichotomising the age variable at 22? Logistic regression can have continuous variables as predictors.
Figure 1: was this analysed using Chi Square? This is not in the Data Analysis section unless it comes under the ‘personal’ information.
Line 223: I suggest re-writing as ‘Results of the multivariable logistic regression are reported in Table 2. Factors associated with high perceived stress in order of highest to lowest odds ratio were; etc etc.  It would be helpful to include the p value in table 2 as per table 2, rather than just asterisk and say ‘Statistically significant’.
It is so helpful to have the data available.  I ran the multivariable logistic regression but obtained slightly different results.  Apologies if I have made a mistake here.  Did the authors include any other variables here that are not in Table 2? Overall interpretation is the same except for Smoking, where in my analysis the OR was <1 (but still non-significant) and Income, which was not significant.
Was there any reason why the dichotomised GPA and Level variables were not included?
Line 229: Heading for this section – the word ‘stress’ needs to be inserted after ‘perceived’.
Line 230: It looks like a t-test has been performed here between male and female PSS scores but there is no mention of this in the Data Analysis. I’m assuming the means and standard deviations are reported.  If a P value is <0.05, please refer to it as ‘statistically significant’ rather than just ‘significant’. While the male and female PSS scores do differ, the effect size (Cohen’s d)  between male and female means – 0.215, or small.  Does a small effect size here mean anything?  The authors have a reasonable sample size here, and so relatively trivial differences can be statistically significant but of no clinical or practical importance.
Likewise most of the correlations in Table 3 are  also small (0.10 = small, 0.30 = medium, 0.50 = large). The effect of the pandemic on health and academic flow and exams are the 2 strongest, the others are relatively weak. The authors said in the data Analysis section that non-parametric correlations were used, so I’m assuming Spearman’s? Please note – Spearman’s coefficient is not known as r (that is for Pearson’s correlation coefficient.  The term ‘r’ is also used in line 237, 240 & 244).  Spearman’s is known as rho.  In Table 3, is the P value for GPA correctly reported or is it the asterisk that is incorrect?  In the table the P value is reported as 0.543, but is asterisked as statistically significant.  By the size of the correlation (-0.023), I’m guessing it is not statistically significant.  It is also not necessary to asterisk the P values and then indicate below the table in text that these are statistically significant – a reader should be able to determine than anything <0.05 is statistically significant.
Please also check the wording of the last 2 questions in Table 3.
Based on the supplied data, I ran the correlations between total PSS score and GPA, and total PSS score and level, and obtained different results.  Please see attachment.
DISCUSSION
Line 254:  Comparing medical students' perceived stress before and during the pandemic showed a significant increase in high perceived stress from 11.6% pre-pandemic to  20.6% during the pandemic.
Please re-write as Comparing medical students' perceived stress before and during the pandemic showed a statistically significant increase in the prevalence of high perceived stress from 11.6% pre-pandemic to  20.6% during the pandemic.
Line 262: Moreover, multivariable logistic regression demonstrated that female students were at a higher risk of high stress during the pandemic than males.
This isn’t actually the case.  The regression showed that females were at an increased risk of high perceived risk compared to males, regardless of the timing.
Line 275: Multivariable logistic regression and the correlation of the factors associated with perceived stress during the pandemic suggest that lower family income is significant in high stress.
Two points here: 1) which variable in Table 3 indicates a relation ship between income and stress? I don’t see income specifically mentioned. 2) Re-write the last part of the above sentence as ‘…lower family income is associated with high stress’.
Line 287: A study conducted during the lockdown and social distancing periods revealed that younger age, besides the female gender and lower income, is correlated with a greater risk of  stress and lower life satisfaction (Rogowska et al. 2020).
Please re-write as ‘A study conducted during the lockdown and social distancing periods revealed that younger age, female gender and lower income, were associated with a greater risk of  stress and lower life satisfaction (Rogowska et al. 2020).
Line 290: t is assumed that the ability of younger students to cope with stress is not entirely mature.
It is not clear what this sentence means. Do the authors mean that due to younger age, students may not necessarily have the maturity to cope with stress?
Line 307: One study reported potential  challenges and limitations in delivering teaching material and clinical training using e-learning technologies. Please provide the reference for this study.
CONCLUSIONS
Line 341: Younger students, those in lower academic  grades, and female students are at risk of having high stress.
The only analysis I can see involving grades (GPA) is in Table 3, and here the association between GPA and stress is not statistically significant.  I attempted to replicate these results (see attachment) and obtained a somewhat different, but statistically non-significant correlation between GPA and stress. I therefore do not see any evidence that lower grades are associated with stress.  Academic level also only appears in the correlations (and is statistically significantly negatively associated with stress) but the relationship is very weak.


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: INCREASED MENTAL STRESS AMONG UNDERGRADUATE MEDICAL STUDENTS IN SOUTH-WESTERN SAUDI ARABIA DURING THE COVID-19 PANDEMIC
Review round: 3
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: GENERAL
I would like to congratulate the authors and responding to the reviewer’s comments and making the paper an improved piece of work.
I have made a number of comments and suggested edits, plus a few things I missed on my first review, apologies.  These are mainly relatively minor.  I did identify that the ‘Income’ variable has not been properly handled in the regression, but this does not affect the interpretation of results in any major way.

INTRODUCTION
2nd paragraph, line 59: recommend deleting the comma after ‘decreased life satisfaction’ and changing to ‘decreased life satisfaction and decreased positive emotions’.
7th paragraph, line 96: delete ‘the’ between ‘among’ and ‘similar’.
METHODS
Study tool
Line 183: ‘…after an extensive review of the literature, a panel of four experts in the…’ delete the comma after literature and insert the word ‘and’.
Line 185: Please state the method for assessing the content validity – was it the Content Validity Ratio (CVR)?
Line 185: “The Cronbach’s Alpha…’ please delete ‘The’, and Alpha can be lowercase (alpha).
Data analysis
Line 200: Please insert ‘Statistics’ after IBM SPSS.
RESULTS
Comparison of high perceived stress levels before and during the COVID-19  pandemic
I now see what has happened with the Income variable. Unfortunately, you have not used the Income variable correctly in the logistic regression.  You have treated it as if it is a continuous variable (1, 2, 3) but it is an ordinal variable, and must be used in this way in the logistic regression, otherwise the coefficients are not correct.  The text (lines 209 – 213) says all predictor variables were dichotomised, but this is not the case for income.
STEP 1: Include the 3 level ‘Income variable’ in the list. Click on ‘Categorical’

STEP 2: Click on ‘Income’ and make sure ‘First’ is selected as reference category.  This is the only appropriate reference category as the last (Insufficient) has too small a sample size to be a reference for which the other 2 levels are compared.

The test will compare ‘Sufficient and exceeds’ to ‘Insufficient’ and ‘Sufficient and exceeds’ to ‘Insufficient’, and you will then have 2 sets of statistics – B coefficients, SEs, aORs and P values, for this variable.
This is what I get if I use the 3-level income variable

The reference is ‘Sufficient and exceeds’ here, and Income 1 = Sufficient, Income 2 = Insufficient’.
So ‘Sufficient’ vs ‘Sufficient and exceeds’ is statistically significant (P = .014). Compared to students with ‘Sufficient and exceeds’ income, those with ‘Sufficient’ are more likely to experience high stress (aOR 1.53 (1.09, 2.15) but ‘Insufficient’ vs ‘Sufficient and exceeds’ is not statistically significant (P = .119).  This is probably due to the sample size in ‘Insufficient’ (n = 28).
If I run the logistic regression WITHOUT specifying Income as a categorical variable, so that the test thinks it is a continuous variable, I get the same results as your table 2. See below.

In your Data analysis section, I recommend writing this as
‘The following predictors of high perceived stress were dichotomized variables: sex (female vs. male), age in years (≤22 vs. ≥23), smoking status (smoker vs. non-smoker) and the timing of the  study (during the pandemic vs. before the pandemic). Income was a 3-level variable as per Table 1, with ‘Sufficient and exceed’ as the reference category.
I understand this changes your argument somewhat regarding the regression, but it is not appropriate to treat an ordinal variable as if it were continuous, as treating it as continuous assumes that the difference in stress between level 1 and 2 is the same as the difference in stress between 2 & 3 and you can’t make that assumption.
You can use the rankings for Income in the correlation because you have used Spearman’s, but there is no equivalent in the logistic regression.
Also please note that the symbol β is for a standardised beta, whereas in the logistic regression it is unstandardised, and so should just be reported as B.
Line 215: It would be helpful to  indicate here the scoring for family income, e.g., ‘income ranked insufficient (coded 3), sufficient (coded 2) and exceeds (coded 1); educational level (1 – 12) and GPA (0 – 7).’  If you don’t want to put them here, I suggest putting them below Table 3, as this helps the reader understand the correlations and they don’t have to remember or read back over previous tables/text.  Also helpful to have rankings for the 8 Covid questions provided, ie Never = 0, Very often = 2).
Factors correlated with perceived stress during the COVID-19 pandemic.
It is appropriate to provide some measure of effect size in any analysis.  I recommend adding in the following.
The average perceived stress scale (P.S.S.) score was statistically significantly (p = 0.005) higher among female students (21.86 ± 6.84) compared to male students (20.46 ± 6.24). The mean difference was -1.41 (95% CI -2.38, -.44).
I note that although there is statistical significance, both means are in the ‘moderate range’.
Line 249: Please remove ‘increased’, you only need to say age here, as the negative correlation implies that as people get older, stress decreases.
Line 252: should be ‘worried’ not ‘worries’.
DISCUSSION
Line 274: There is a fullstop after ‘pandemic’ that needs to be removed.
Lines 279 – 282: Our results showed that high stress was more prevalent among female students compared to male students both before and during the pandemic. Moreover, multivariable logistic regression verified that female students were at a higher risk of high stress.
The t test reported in lines 248-249 showed that female mean stress higher than males during the pandemic.  Note - means do not indicate prevalence. No t-test of means between males and females pre-pandemic was reported. The regression showed that, irrespective of pre or during, females are more at risk of higher stress.  I therefore think you do not need both these statements – just a statement that irrespective of the timing of the study (pre or during the pandemic) females are at risk of higher stress than males.
Line 308: It is assumed that the younger students face difficulties coping with stressful conditions.
This is a big assumption to make.  I also think it is unnecessary, as the preceding sentence and the Rogowska reference covers this issue.
Line 318: A fear of psychological distress is a prominent factor for developing stress  and anxiety associated with crises, especially among university students.  Do the Chau et al and Huskey et al include this information about university students?  Otherwise a reference is needed for this.
Line 328:  In the present study, worries about the possible disruption of academic lessons and exams were a major source of stress among medical students.
Remember, correlation is not causation!  There was moderate correlation between stress and academic disruption, so all you can say is
In the present study, worries about the possible disruption of academic lessons and exams were positively correlated with stress among medical students.
Likewise in line 335 – replaces ‘sources of stress’ with correlated with stress.


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: FLOTILLIN-1 PROMOTES EMT OF GASTRIC CANCER VIA STABILIZING SNAIL
Review round: 1
Reviewer: 1

Output:
Basic reporting: The overall writing quality of the manuscript is good. Nevertheless, some problems need to be solved :
-	Line 40 : replace « protein and implicated » by « protein and it is implicated »
-	Line 117-118: correct the sentence. « Together those results shown that Flotillin may play an oncogene in the development of gastric cancer ».  May play an oncogene is too strong. « can participate in the development of gastric cancer ».
-	Line 303 : Flotillin 1, 1 is missing
-	Figure 1B : replace T by tumor and N by normal.
-	Figure 1D : explain what are the T stages.
-	Figure 3 shRNA and not only sh has to be written on the legend
-	Line 143-144 : the sentence has to be modified, « but has no effect on the mRNA level ».
-	Line 160 : the sentence has to be modified, with tumoral cell progression.
-	Lines 170-171 : the sentence « Therefore, our results demonstrate Flotillin-1 play a key oncogene in the development of gastric cancer, especially in promoting metastasis » has to be modified. « Therefore, our results demonstrate Flotillin-1 participate in the development of gastric cancer, especially in promoting metastasis » is one possibility.
-	Lines 172-182 :  the quality of the writing is not good and this part must be rewritten.
-	Line 183 : « that » is missing after identified.
-	Line 184 : there is a mistake «  downregulated mesenchymal markers » has to be replace by  « upregulated mesenchymal markers ».
-	Line 187 : « that » is missing after demonstrates. The sentence « promotes gastric cancer metastasis through inducing EMT » has to be tuned down, the proposition is  « and it overexpression could participate in… ».
-	Line 208, again oncogene could not be used. The sentence has to be modified.
Experimental design: The experiments are globaly correctly conducted. Nevertheless, some points need to be addressed. They are detailed below:
1- Control cell lines.
-	In figures 2, 4 and 5, using control cells that are the parental cells is not correct. Indeed, the cells (for both cell lines) that overexpress flotillin 1 need to be compared to cells that express a control plasmid.
-	In figure 3, using control cells that are the parental cells is not correct. Indeed, the authors have to use a cell line expressing a control shRNA.

2- Explain the cell line generation (both the one that overexpress flotillin 1 or shRNA Flotillin 1). I could not find the information in the material and methods section. This has to be explained. I was wondering whether stable cell line expressing flotillin 1 were generated/used or if the overexpression of flotillins was performed by transient expression ?

3- Quantification of the western-blot data.

Any quantification is shown on the western blot. This has to be done on independent experiments and it is required in order to evaluate the reproducibility of the data.

4 - Other EMT-inducing transcription factors
What about the expression of the other EMT-inducing transcription hy the  ? Why the authors analyzed Snail and not the other ones such as Zeb1/2 or Slug?

5 - Flotillin 2 analysis
-	Figure 2, the level of flotillin 2 is important to monitore by western-blot since these proteins work together. Is the overexpression of flotillin 1 associated to an increase in flotillin 2 ?
-	Figure 3, the level of flotillin 2 is important to monitore by western-blot since these proteins work together. Is the knock down of flotillin 1 associated to a decrease in flotillin 2 ?
Validity of the findings: Some modifications are needed:

1 - Quantification of the western-blot data. (already mentioned above in Experimental design)

Any quantification is shown on the western blot. This has to be done on independent experiments and it is required in order to evaluate the reproducibility of the data.

2- Some sentences are to be modified (detailed in 1: Basic reporting). IN particular saying the flotillin 1 is an oncogene is not possible. The authors express flotillin 1 in transformed cell lines harbouring already oncogenic events.
Additional comments: none


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

Input:
Title: FLOTILLIN-1 PROMOTES EMT OF GASTRIC CANCER VIA STABILIZING SNAIL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Huang et al. provide information on the role of Flotillin-1 in gastric cancer cell lines expanding the knowledge on the involvement of this protein in tumor biology. The manuscript is clearly written, the introduction gives the necessary information, and the references are relevant. The results are interesting. They show how the modification of Flotillin-1 expression (by overexpression or downregulation) affects  migration and invasion of gastric cancer cells.They have studied the expression of some markers of EMT and they have found that flotillin-1 regulates the protein expression of Snail without modifying its mRNA level, likely by increasing its stability. Some modifications in the text are necessary before acceptance of the manuscript. The manuscript might be improved giving more information on the experimental conditions, including the number of replicates in the main text. Most of the conclusions are well stated. However, since only "in vitro" experiments have been performed it should be considered to smooth some of the statements (see below). The demonstration of a role of Flotillin-1 in metastasis formation should require injection of Flotillin-overexpressing and control tumor cells and measurement of the appearance of metastasis "in vivo".

.

.
Experimental design: Methods: The secondary antibodies used are not listed in Methods. No information about how many cells were seeded, or how many micrograms of protein were loaded for western blot is provided. Information on how many independent experiments and how many technical replicates in each experiment should be provided in the main text.
Validity of the findings: Most of the conclusions are well stated. However, since only "in vitro" experiments have been performed it should be considered to smooth some of the statements (see below). The demonstration of a role of Flotillin-1 in metastasis formation should require injection of Flotillin-overexpressing and control tumor cells and measurement of the appearance of metastasis "in vivo".

127 Taken together, the results show that overexpression of Flotillin-1 promotes gastric cancer metastasis.
Since only cell culture experiments have been performed, I suggest to change the sentence: the experiments show that overexpression of flotillin promotes cell migration and invasion and suggest that flotillin might have a role in cancer metastasis.
The same applies for 135-136:Thus, the results show that knockdown of Flotillin-1 inhibits gastric cancer metastasis.
Additional comments: Sentence not clear:
118, 170: “shown that Flotillin-1 may play an oncogene in the development of gastric cancer”.

Figure 1B: Please indicate the meaning of TPM
Figure 1C Please indicate the meaning of HR and the meaning of the different black and red numbers under the X axis. Please make clear in the text the provenance of the samples of tumors expressing low-Flotillin and high-Flotillin, used to build the plot.
Figure 1D Please specify the meaning of RSEM in the axis .

148, 201: I do not understand what the Authors mean with “liable protein”. Do they mean labile?
348 Figure 5: " Flotillin-1-knockdown and control SGC-7901 cells were treated with CHX as indicated".
The legend of the figure contradicts the main text. Do the Authors mean Flotillin-1-overexpressing and control SGC-7901…
 It is difficult to understand why control cells do not express Flotillin, while cells labeled as “Flotillin”, which are supposed to be knocked-down for the protein, express it.

Abbreviations:
The authors should provide the information of the meaning of the abbreviations used. It should be convenient to indicate that CDH1 is the gene coding for cadherin 1. HMT is abbreviated but it is used only once. The meaning of  DNMT, HDAC  should also be provided.


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: FLOTILLIN-1 PROMOTES EMT OF GASTRIC CANCER VIA STABILIZING SNAIL
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors reported that flottilin-1 inhibit ubiquitination of Snail in gastric cancer cells to promote EMT.
The fugires are clear and experiment are well done.

I have two minor comments.

1. The authors declared that gastric cancer is included in 23 cancer series, but I didn't find it. Please let me know the data which shows gastric cancer has amplification of flotillin-1.

2. The authors insisted that ubiquitination of Snail was inhibited by flottilin-1. What is a deubiquitinase involved?
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: No additional comments


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

Input:
Title: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper described the effects of seasonality, fertilization and grass species on nitrogen-cycling microbial abundance and transcriptional activity. It is well written and provide evidence for ecological importance of moderate fertilization.

My biggest issue is that the paper contains too many previously published data and very often did not present them as such (even authors did not try to hide this fact). The paper should centre on the new data (denitrification part) and use other data mostly for comparison and explanation purposes. If the authors have to re-describe the old data, please at least cite it properly (including in the figures and captions).
Experimental design: no comment
Validity of the findings: no comment
Additional comments: (1) Line 107-109. What are the thresholds for “moderate” and “high” fertilizations here to generate this hypothesis? Does your introduction contain such message? For instance, the authors mentioned “250 kg N ha-1” fertilization increased soil denitrification exponentially with rate (line 84-85), isn’t that high fertilization rate range which should decrease such activity? And why should moderate and high fertilization have opposite effects? Toxic effect of excessive N? The authors should make it clear in the introduction.
(2) Line 154 and 155. I cannot see the formula
(3) Line 181-197. I cannot tell if the section is original analysis or already presented in other papers.
(4) Line 242-244. Is there statistics for this conclusion?
(5) Line 248-249. It might be true in this study but what is the indication here? Should we give up assessing gene expression from now on, since it does not correlate well with the soil properties and biochemical processes? Please discuss this thoroughly.


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: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: The first sentence of results sends me to another paper... this is a recurring issue for this paper. I'd appreciated it if the authors could reframe the story. For example, there are numerous papers in the long term ecological research (LTER) system are working on previously published studies, and while it is of course essential to acknowledge that, those papers tend to put their new findings and the novel research components at the center of their results and discussion. I think this manuscript would greatly benefit from a narrative that is focused on their novel contributions. As is, I feel like it would have been great to include the new data presented in this paper, in their original paper. I recognize there are many logistical and practical reasons why the authors may have chosen to parcel up the research, but the flow of this paper and its current form come across as incomplete.

Bottom line, since this paper should adhere to the novel research questions and findings component of original scientific research, then the authors need to revisit how they have structured this paper to better highlight their unique contributions.
Experimental design: Lines 107-11: Please provide your reasoning for why you expect this hypothesis to be true (in H2: you say because of the different optimum N rate for these species, but magnitude and directionality are helpful here. Please clarify why you came to these conclusions or rewrite the introduction to lead the reader to these hypotheses as well.

Methods: line 130: Please be more detailed and specific about frequency and amount of fertilization and the timing of fertilization. I see them listed above in the N application rates sentence, but were they all applied on the same day? which seasons? how frequently? Are the April and June timings listed below consistent for every year starting in 2013? I am really confused, which inhibits clear interpretation of the data which also inhibits proper comprehension of the discussion section too.
Validity of the findings: Section 2.2 reads as highly unusual.
It feels like you are stating this paper doesn't merit the detailed methodology for about half of the data of this paper. Which seems problematic as the methods section should be fully reproducible.

The authors mention the dispersion visualized by PCoA but didn’t elaborate or interpret it further. Please clarify in the discussion.

I was generally confused and kept needing to return to the methods section, but am not sure in some of the results because of a lack of clarity in the methods.
Additional comments: This is really interesting research, and exciting to see the parallel work on functional genetics and transcriptomics. I look forward to seeing the writing improve the clarity of your research.

The last line of the abstract left me expecting a different type of discussion section, one that would focus in on the suggested functional stability of N cycling populations in native C4 grassland systems. I would encourage the authors to elaborate on that final thought from their abstract, or revise the abstract to better match the narrative of this paper.

Line 88: can you expand on potentially why this high variability was observed? Provide additional context for the readers please.

Line 99: please be explicit by mentioning in what ways N-cycling microbes might be affecting N rates.

Line 101: please list off the limited studies you reference here and cite them. Explain what those limited studies have contributed to help contextualize the work beyond your previous papers on this same study site so the reader can have a broader understanding of the importance of this research.

Methods
Statistical analysis: Grateful to see fixed effects specified here. Would be helpful for me to see the total sample size for your statistics as well.

Statistical tests could be run in R rather than in SAS and R, which would make the output of the paper cleaner and allow to better reproducibility.

Results
Sections 3.2 and 3.4 lack any references to statistical tests or p-values, please provide these in text where appropriate.

Conclusions
I think building off of pre-existing experiments is excellent and important work, but this paper doesn't stand on its own, which is a general requirement for scientific papers. I am excited to see the authors digging into functional potential and transcript-level responses, that is really exciting work! But I don't see that being highlighted here, and I would love for the authors to showcase this work and its unique contributions to our understanding of grassland systems.

In general there are a few places where I need greater clarity in writing style to follow the author’s narrative. Please revise with increased clarity in mind.


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: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: Dear authors,
I enjoyed reading your manuscript and believe that it meets the standards of PeerJ and should be published. What I really liked is that you made your qPCR results publically available as this is not done very often.
Experimental design: I only have to minor suggestions:
L144 to 145: the authors should briefly state how they performed real-time PCR.
L154 to 155: the equasitions are not displayed in the Review PDF.
Validity of the findings: The authors performed statistically robust sound and robust tests.
Their conclusions are well stated and linked to their orignal research questions.
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: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 2
Reviewer: 1

Output:
Basic reporting: I am satisfied with the authors' response to my questions and the manuscript is greatly improved after the revisions. Therefore I support the publication of this study.
Experimental design: NA
Validity of the findings: NA
Additional comments: NA


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: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript has been improved, but typographical issues still crop up on and some additional clarification points.

In some sections another read through of the tracked changes version is needed (e.g. fertilized and harvest should read fertilized and harvested in line 182). In other areas, the revised sentence still leaves this reader confused (e.g. lines 241-244 when referencing absolute vs. relative measurements of 16S. The authors wrote "... we measured" where they measured both relative values in the current study and absolute values in the former. I am still confused which one they mean in this sentence).
Experimental design: There are still areas where a method is stated but no citation follows, again mostly throughout the methods section. For example: "Berthelot reaction-based and Vanadium (III) chloride-based spectrophotometric methods" In an effort to make this study stand alone as a paper a citation is still needed as you've done in many areas when referencing the original article, but would help this reader follow along with which methods are bespoke to your lab or modifications from standard protocols. In either case I'd appreciate the reference or details for each method, as it helps clarify exactly what measurements were taken and how you went about those procedures.
Validity of the findings: Some statements are terse and lack explanation, even when in the discussion section (this approach reads completely fine in results which should be straightforward, but this reader prefers author interpretation or citation for claims that aren't explicitly supported by this study's data). For example: "We found that 202N increased N2O emissions. One of the possible reasons is the lower pH caused by high N fertilization rate." which the authors follow with a citation explaining one possible reasoning for the lower pH connection, but the mechanism isn't spelled out for readers.
Additional comments: No additional comments


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

Input:
Title: UREA FERTILIZATION AND GRASS SPECIES ALTER MICROBIAL NITROGEN CYCLING CAPACITY AND ACTIVITY IN A C4 NATIVE GRASSLAND
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors addressed all my comments and concerns well. I recommend their work for publication.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: A NEW MODEL FOR FREEDOM OF MOVEMENT USING CONNECTOMIC ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript is in general well-written and easy to follow. The tables and figures are thoroughly described, and I thought Figure 4 was well-designed.
Experimental design: The authors adopt a causal view of volitional action as a sequence of steps. Then they identify brain regions associated with these functions from the BrainMap database of prior fMRI studies. Next, they try to perform an abstract “tract tracing” through these functionally defined regions using “tracts” identified from the Brainnetome database with a graph-theoretic interpretation. This network is then interpreted as the key network for voluntary action.

In a very general sense, I see the merits of the author’s intentions. Prior imaging studies with fMRI often focus on describing a cognitive ability as a functional network, which is effectively a list of brain regions that are engaged in tasks involving that cognitive ability. However, relating these regions to actual structural connectivity constraints is an important next step, especially using the new publicly available connectome databases. Although this is an important goal, in my evaluation, the author’s execution of this goal seems a bit too preliminary and overly simplistic in its current form.

MAJOR CONCERN 1
In general, the issue of how all the pieces fit together for voluntary action is important. However, I’m having difficulties in determining whether this manuscript is a connectome-oriented (i) review, or (ii) quantitative meta-analysis, or neither.

Prior reviews on this topic have sought to provide integrative overviews for voluntary action. Their objective is often to provide a conceptual scheme to integrate heterogeneous considerations and findings. Other studies, for example with Activation Likelihood Estimation (ALE) seek to quantitatively integrate the heterogeneous findings from different experimental studies to identify commonalities and, importantly, to statistically assign a confidence to these commonalities.

The current study does not seem to be a quantitative meta-analysis. [See, for example, Zapparoli, L., Seghezzi, S., & Paulesu, E. (2017). The what, the when, and the whether of intentional action in the brain: a meta-analytical review. Frontiers in human neuroscience, 11, 238.]. In Table 1, the evidence for each “module” and the associated brain regions is based on isolated studies and there is per se no analysis of whether empirical studies of voluntary action are consistent with these regions. The assigning of function to brain activity is based on associations from studies on other topics obtained from Brainmap rather then volition itself. For example, consider BA11’s role in "motivation".  Might BA11 be broadly associated with emotion and cognition? Yes, possibly based on studies on different topics. However, is BA11’s role in voluntary action related to motivation? This is a far more complex, ambiguous answer and is not addressed.

In summary, there is little evidence provided for the reliability of the different regions and their functions listed in Table 1, in studies specifically about voluntary action. This is an issue that needs to addressed by the authors. Are the regions listed in Table 1 (1) identified based on broad conceptual considerations of general function, or (2) from quantitative regularities observed in studies explicitly about voluntary action? Without this clarification, the regions listed seem quite ad hoc.

MAJOR CONCERN 2
The authors describe a causal model in Figure 1 which is central to the entire study. In a logical sense, the steps are reasonable (motivation, what/how/whether/when to move, etc.). They are decisions that must necessarily be involved at some point before an action. However, the entire study is based on a strict interpretation of the order in which these steps occur (Figure 2). This is an extraordinary simplistic assumption and one that is indefensible without context.

To provide a simple example, I can decide today that I will move one of my fingers at 10am tomorrow (when) and postpone the decisions about which finger to move, by how much and whether to do actually do it until the time arrives. This is a challenging aspect in Libet’s studies.

The more serious concern is the extent to which brain networks could be assigned to these causal functions.  For a visually guided action, classical psychological models suggest that there is an ordering of events. [seen]=>[identified]=>[response selected]=>[executed]. These are logically necessary. However, their NEURAL implementation and sequencing poses an entirely different issue. A minimum requirement is that the stimulus be sensed and the action be executed. So, information has to putatively travel from the visual cortex until the motor cortex where signals to execute the action can be discharged through the cortico-spinal tract to the effector. However, the ordering of these steps in the brain and whether they actually occur in such an order has been anything but controversial. For instance, it does not preclude that the responses are prepared BEFORE the stimulus arrives.
•	Cisek, P., & Kalaska, J. F. (2010). Neural mechanisms for interacting with a world full of action choices. Annual review of neuroscience, 33, 269-298.
•	Ploran, E. J., Nelson, S. M., Velanova, K., Donaldson, D. I., Petersen, S. E., & Wheeler, M. E. (2007). Evidence accumulation and the moment of recognition: Dissociating perceptual recognition processes using fMRI. The Journal of Neuroscience: The Official Journal of the Society for Neu- roscience, 27(44), 11912–11924. https://doi.org/10.1523/ JNEUROSCI.3522-07.2007
•	Viswanathan, S., Abdollahi, R. O., Wang, B. A., Grefkes, C., Fink, G. R., & Daun, S. (2020). A response‐locking protocol to boost sensitivity for fMRI‐based neurochronometry. Human brain mapping, 41(12), 3420-3438.

In summary, the strict ordering assumed by the authors is highly problematic. It would be important to assess the extent to which the results rely on this sequential progression. How similar would the results be if the ordering of the steps were different? Also, why is the assumption of sequential ordering even necessary when considering the structural connectivity? If no assumptions about ordering are made, how much would the conclusions  about connectivity differ? For instance in Figure 3, if the connectivity matrix is considered without restrictions on the steps then there would be a much higher number of edges. How much does this effect the interpretation?

MISCELLANEOUS
There are several ambiguities in discussing connectivity. From Lines 127-137, it is unclear whether the connectivity is based on structural or functional connectivity. What if functional connectivity were to be considered for the same regions? Why would this not be a suitable view since voluntary action is also a functional pattern of neural activity

How do these findings relate to the connectivity conclusions of this work → Zapparoli, L., Seghezzi, S., Scifo, P., Zerbi, A., Banfi, G., Tettamanti, M., & Paulesu, E. (2018). Dissecting the neurofunctional bases of intentional action. Proceedings of the National Academy of Sciences, 115(28), 7440-7445.

The authors might want to consider looking into the Brain Connectivity and Behavior toolbox (Michel de Schotten) [http://toolkit.bcblab.com/] that has sought to do what the authors are doing for lesion analysis. Specifically, given a lesion, the toolbox provides a means to evaluate the tract passing through that lesion to identify cortical regions connected to that lesion.
•	de Schotten, M. T., Foulon, C., & Nachev, P. (2020). Brain disconnections link structural connectivity with function and behaviour. Nature communications, 11(1), 1-8.
•	Nozais, V., Forkel, S. J., Foulon, C., Petit, L., & de Schotten, M. T. (2021). Functionnectome: a framework to analyse the contribution of brain circuits to fMRI. bioRxiv, 2021-01.
Validity of the findings: With the issues listed above, the validity of the findings are low and the interpretation in the Discussion considerable exceeds what might be reasonably concluded from the findings.
Additional comments: No additional comments


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

Input:
Title: A NEW MODEL FOR FREEDOM OF MOVEMENT USING CONNECTOMIC ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Major
1. Much of the manuscript is written clearly with appropriate references throughout. In the Introduction, the description of Hallett’s neuroanatomical model of volition (Lines 45 - 80) could be reorganized to improve clarity and remove redundancy. It would also be helpful to describe some of the empirical evidence for the model (i.e., why is pre-SMA implicated in movement timing?)
2. The labeling of Brodmann areas are sometimes inaccurate, which raises concerns about the interpretation of results. For example, the authors equate BA6 with pre-SMA, but BA6 more generally encompasses SMA and premotor cortex in addition to pre-SMA. Likewise, it is not clear that BA14 is part of the insula in humans. In general, the use of Brodmann’s areas in the text is sometimes confusing—I think it would be easier to conceptualize the model if the authors used appropriate anatomical labels along with the description of the BAs or sub-areas (ex. Line 206 could be modified to: “since BA9m (dFMC) has been implicated in self-initiated (conscious) inhibition of movement”)

Minor
1. The phrase “idea of liberty” seems out of place here, as the conceptual model is specific to the study of action, and it is unclear if it generalizes to other cognitive domains.
2. There are some awkward sentences and grammar (e.g., Lines 40-41 “We feel capable of deciding our actions and execute (or stop) them”; Line 31 “less connections”) and colloquial language (“we checked”/“we wondered”).
3. Why does the step organization start at Step “0”? In addition, could the Steps with an inhibitory component be labeled “a” and “b” (e.g., Step 5a?)
4. Fig. 2: Add legend for color coding the different steps.
5. Fig. 3: This figure is low resolution, which made it difficult to discern region names
Experimental design: Major
1. The research question and identified knowledge gap could be more strongly motivated. As currently written, it is not obvious how the present study extends previous neuroanatomical models of volition other than mapping out the network in more detail. Instead, a stronger emphasis could be placed on the utility of a connectomic approach for (a) confirming that Hallett (2016) model is plausible, in terms of human structural connectivity, and (b) identifying smaller sub-regions within the broader volitional network that may serve as critical nodes to investigate in future studies.
2. If the goal was to perform a connectomic analysis, I do not understand why the authors chose to simply “count” edges rather than use graph theoretic measures well-described in the literature (Bullmore and Sporns, 2009). The authors make inferences about complex networks concepts, such as “hubs”, “rich club”, and “robustness”, but these can be actually quantified by assessing node degree, clustering coefficient, centrality, etc. With the current approach, these designations about network properties seem only speculative.
3. I am struggling to follow the logic of the network analysis, as detailed in the Results, Fig. 3, and Table 2. First, many of the results seem to be a direct consequence of the regions selected for each step, rather than revealing anything in particular about the properties of the volitional network. For example, “For the inhibitory component of step 0, the dorsoposterior cingulate cortex (BA23) is the most connected node with 79.28% of total bilateral edges, leaving the ventroposterior cingulate cortex (BA31) with only 20.72%” (Lines 174-176). But this difference can be explained by the fact that six BA23 subregions were included (right and left BA23d, v, and c) that have edges with each other, relative to only two BA31 subregions. Of greater concern, the authors state that “The insula module had 100% of edges arriving from nodes active at step 4, suggesting it is the only area capable of generating positive agency” (Lines 232-234). This results seems to stem from the decision to only include insular cortex subregions in that step’s analysis. If this is the case, how can the edge contribution of the insula be anything other than 100%? The argument appears to be circular.
4. Lines 130-137: There needs to be a more complete justification for the selection of regions from the Brainnatome atlas. How exactly did the authors select the regions to be included in each step?
5. The connections between regions in Figs. 1 & 4 are very nicely visualized (especially Fig. 4). However, it should be more clear which aspects of the model are assumptions from previous work vs. results from the present analysis. For example, the Figures show inhibitory and feedforward connections between regions, but there is no way to know this from the Brainnatome connectograms alone.

Minor
1. Fig. 3: Label whether each sub-region is right or left hemisphere. Also, it does not make sense to color the matrix elements outside the black boxes blue, as this would imply that there is no edge between the regions (even though connections outside the black boxes were not assessed). In general, consider a different visualization approach for Figure 3. It might be helpful to try a circle plot visualization (Irimia et al., 2012) separately for the different steps.
2. What is meant by “this area is not present in the right hemisphere”? (Line 182) and “lack of BA39 in the left hemisphere” (Line 247). Please clarify.
3. It is unclear what BrainMap adds to the analysis—why is it important to list the various functions of a particular region if it is not relevant to volitional control?
Validity of the findings: Major
1. Related to the comment above, I find the conclusions about the percentage of edges (e.g., “Indeed, 78.96% of edges happen during the first three steps.” Line 286) problematic given that they could be mainly driven by the number of regions selected for the different model steps. The interpretation is interesting (delineation of early flexible motivation and selection processes vs. later simpler initiation and agency processes), but not convincingly supported by the results.
2. Likewise, the discussion of implications of the model for neurological disorders is interesting but beyond the scope of the study, especially since similar ideas were discussed in previous work (Kranick & Hallett, 2013). These sections could be removed or condensed significantly.
3. One of the central claims of the study is that “From the structure of this model, we propose that consciousness is not required for movement generation” (Abstract, 33-34). I do not really understand this point and I am not sure how the presence or absence of conscious processing can be determined from the model.
Additional comments: The overall idea and motivation for the study is interesting and could have important implications for future studies on the volitional control of action and sense of agency. My main concern is the emphasis on utilizing a connectomic approach without taking advantage of established measures for graph analysis of brain connectivity data sets.


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

Input:
Title: A NEW MODEL FOR FREEDOM OF MOVEMENT USING CONNECTOMIC ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I would like to commend the authors for the considerable amount of work done to revise the manuscript. I think this revision has vastly improved the manuscript’s content, rigor and clarity. All my major concerns have been sufficiently addressed.

Supplementary Table 1 is important but was missing from the review materials.

A minor suggestion is that the labeling of the figures and the captions could use improvement as they contain a lot of important information.

Figure 2: The caption is a bit too sparse to describe the figure.
•	The panels 2E and 2F do not seem to be referenced anywhere in the text
•	It could be useful if the colors of the nodes in the graph correspond to the colors in Figure 1 (as is nicely done in Figure 4) to relate these measures to the different "tasks".

Figure 3 is interesting and important but it was not possible to see any details even when zooming into the pdf. This seems to defeat the purpose of displaying the figure. Maybe the authors could consider providing high-resolution vector graphics images of each of these connectograms in the Supplementary Material so that the labels and values could be examined by readers. The authors could add the name of the task (e.g., “Motivation”, “Agency”, etc.,) next to the corresponding connectogram to make it easy to understand this figure.

Figure 4 –  a legend with the color and task could be added to the figure so that the reader does not have to keep referring back to Figure 1 to find out which task corresponds to each color.

Figure 5 seems relevant after Figure 1 rather than helping to organize the information from the quantitative graph theory analysis. A suggestion could be to move it a position after Figure 1, and before the numerical results.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


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

Input:
Title: A NEW MODEL FOR FREEDOM OF MOVEMENT USING CONNECTOMIC ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revised manuscript is well-written, logically organized, and much improved.

Minor comments
1. Some of the details of the network analysis presented in the Results should be moved to the Methods (e.g., computation of small-worldness, lines 225-240)
2. I am not sure "Tasks" is the best word to use for the model description. Perhaps "Processes" is more appropriate? Additionally, referring to 6 tasks throughout the manuscript is a bit confusing, as sometimes 8 separate results are presented (e.g., the 6 tasks + 2 task modulators).
3. I preferred the naming scheme of "How", "When", and "Whether" (or similar) to "Planning", "Timing", and "Decision." The latter, introduced in this revision, typically encompass a broader set of processes than implied in the manuscript (e.g., "Decision" is commonly used to refer to selection of a particular action goal, effector, etc. rather than the go/no-go decision described in the Introduction).
4. It would be good to reinforce that the analysis is based on structural, not functional, connectivity throughout the manuscript (one example, line 138 "Here, we identify the *structural* connectivity network")
5. Figure 2 is visualized nicely; however it is difficult to appreciate without referencing the Supplemental Table (also true for Fig. 4 to a lesser extent). At the least, it would be good to color code the nodes to its corresponding Task, as is done with the other Figures. Additionally, since 2E and 2F are redundant with 2D, they should be removed.
6. There are a few instances where the old "Steps" terminology is used (e.g., Lines 478, 500, 508-510).
Experimental design: I appreciate the authors' extensive revisions and new graph theoretic analyses. The approach is much improved and addresses many of my previous concerns. However, some concerns with the selection of regions remain, along with additional concerns with the new results.

1. The results of the connectivity analysis obviously rests on the selection of regions. Although the Introduction now provides more information supporting their choices, I still find the region selection is somewhat arbitrary and lacks strong justification, for reasons detailed below:
    a. The selected regions do not appear to fully align with Hallett's model, missing regions in some cases (e.g., hypothalamus, non-limbic basal ganglia) and adding regions in other cases (e.g., insula for Agency--my understanding is the Hallett model mainly identifies parietal regions, and that the sense of agency arises from parietal-premotor connections).
    b. Though it is reasonable to include regions that extend Hallett's model based on other literature, the the decision to include additional regions is inconsistent. For example, the authors cite a study showing posterior cingulate activation during extrinsic motivation to act (Lee et al., 2012). In a follow-up study (Lee & Reeve, 2013), the angular gyrus was activated in extrinsically motivated contexts, yet this region is not considered for Motivation. Similarly, Nahab et al. (2011) found many regions, including supramarginal gyrus, precuneus, dlPFC, and cerebellum implicated in Agency, in addition to the superior temporal gyrus referenced and included in the model.
    c. I am curious as to why the authors did not constrain their regions for the Planning, Timing, and Decision Tasks from meta-analyses of volitional action  (e.g., Zapparoli et al., 2017, as mentioned previously by Reviewer 1).

Overall, I do not necessarily think the analyses need to re-done with new regions, but there needs to be a much stronger rationale for the approach to region selection. The exclusion of the classically "motor" regions of the basal ganglia (e.g., dorsal striatum) is particularly concerning, given that it may share a similar connectivity profile to the nucleus accumbens, which was identified as a principle node in the network in the Results.

2. All BA6 sub-regions are included in the Planning and Timing tasks, which is reasonable, however, as mentioned in my previous review, these regions extend beyond Pre-SMA. It is better characterized as Pre-SMA/SMA/Premotor cortex, and any Results with BA6 should reflect potential contributions from each of these regions rather than just Pre-SMA.

3. Some of the selected regions were restricted to one hemisphere (e.g., right BA39), yet in the model all regions were included bilaterally. Why is this the case, if left BA39 for example, is not considered to be part of the volitional network? How does this influence the hemispheric differences found in the connectivity analysis?

4. Figure 5 seems out-of-place in this revision, as the visualization seems to be mainly based on previous literature rather than incorporate connectivity findings from the present results. For example, BA4 is shown to have connections to the insula and BA6, yet the Results for Execution (Fig. 3G, Lines 316-17) show many more connections between BA4 and other areas. The nucleus accumbens, which features prominently in the Results, is not included. The figure may also be misleading in its depiction of a sequence of steps, rather than reflecting the undirected analysis performed.

Minor
1. Why is the amygdala attributed to Agency (in addition to Motivation) in Figure 4 and the supplemental Table?
2. Much of Discussion section 3.4 (Line 476) is no longer supported by the current Results. In particular, the statement "We found that without the insula the entire mismatch system becomes ill-connected since, with its absence, other areas would be rendered isolated" refers to the previous version of the manuscript.
3. The insula is not discussed with respect to Agency until the Discussion (Line 479). Please provide rationale in the Introduction.
4. Line 157. Do you mean "Areas thus identified are at the level of Brodmann areas or *subcortical areas*"?
5. Could the authors provide a reference for that notion that a network is small-world if Sigma > 1?
Validity of the findings: The validity of the findings would be improved with a much stronger rationale for region selection for the different Tasks of the volitional model.
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: MYTHS IN PSYCHOLOGY: PSYCHOLOGICAL MISCONCEPTIONS AMONG SPANISH PSYCHOLOGY STUDENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: While I think the core premise of the study is interesting- comparison of myths in psychology majors from different types of university settings and in a country that has not been previously investigated but these central contributions are not as clearly stated and addressed.
For comparison of university types-Throughout the paper this should be referred to by their features not the specific school name (online vs traditional).  In results the range of ages are reported and seem to overlap similarly so the argument that this is a different population is not as clear as suggested in the introduction.
I find the concept of scientific dissemination a confusing construct and only measured by one yes/no item that it is not clear students understand. The issue is not scientific dissemination itself but how accurate that dissemination is and being able to critically evaluate the source.  It is essential that science be disseminated to a larger audience than just scientific journals.
It is only mentioned briefly and not with regard to specific studies but important to discuss the countries in which other data has been collected when making the case to transfer to Spain- clarify if this has been looked at non-US samples previously and how that data compares (you have references from London among others but don’t address this explicitly).

There is awkward wording throughout sections of the paper.
For example, the first sentence of the introduction is confusing and does not translate well into English.  The first sentence of the abstract is much clearer and related to the topic you are addressing.
Line 41- the word “proven” is not substantiated
Line 44-46 confusing sentence
Line 58-59 clinical areas would include therapies without empirical evidence
Line 67- need a secondary citation for work in 1920s
Line 92-93 awkward wording of sentence
95-97 range cited is very wide (and is there really a cap to the hours?)
Statistical tests and numbers should not be included in discussion
Experimental design: See note above about clarity of research question and concern about meaurement of "scientific dissemination"
Measures- what criteria were used to eliminate or add items from the existing measure?  Mention “irrelevant” but is that based on numbers reported in prior studies?
Raw data supplied.  Question about last column (in Spanish) appears to list careers/majors but that does not seem to match the way the sample was described as psychology students (unless it is career goals for seniors?)- no column heading
Study is not "observational" (line 287)
Validity of the findings: When reporting a difference between means, need to include the test that supports that statement (line 313 and 325, 363 as example)
line 343 needs to be new paragraph
Clarify more in comparison to other research impact of "don't know" and whether or not that is included in prior studies (some have, some do not)
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: MYTHS IN PSYCHOLOGY: PSYCHOLOGICAL MISCONCEPTIONS AMONG SPANISH PSYCHOLOGY STUDENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Thank you for allowing me to participate in the review process of this study.
The present study analyses the prevalence of false beliefs or psychological myths in Psychology undergraduate students at two Spanish universities. In addition, it analyses how variables such as academic year or familiarity with scientific knowledge media could modulate the prevalence of these false beliefs.
I believe that the study addresses a very important and topical issue. Myths or false beliefs about academic or professional content are a problem in the scientific/academic advancement of scientific areas, so knowing the prevalence of these false beliefs and the variables that could help us to combat them is an interesting contribution to the scientific community.
The paper shows a well-organised theoretical background, and I believe that it helps both the novice and the more experienced reader to contextualise and justify the present study. It also clearly addresses potential methodological problems. The analysis of biases in the use of the tool is very illuminating.
Experimental design: See attachment
Validity of the findings: See attachment
Additional comments: See attachment


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: MYTHS IN PSYCHOLOGY: PSYCHOLOGICAL MISCONCEPTIONS AMONG SPANISH PSYCHOLOGY STUDENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I still find the concept of scientific dissemination difficult- especially as measured here by the single item.  While the letter addresses the concept, I did not see that change in the paper and found the examples confusing- especially as it appears item specifically states not a search in database for academic paper and that is the most frequently reported?  (line 341 results)

I find the label "not face-to-face" confusing for school and think either online or distance learning would be more helpful distinction.

In abstract and at times in paper, phrasing sounds longitudinal but comparing different students in different years (courses is awkward wording across countries)

In respsonse to feedback acknowledged international component but not brought into intro
Experimental design: `See comments about scientific dissemination above and in addition may be more helpful to conceptualize as scientific vs pseudo-scientific sources.  In US many school incorporate information literacy directly which should give skills to students in general before getting to PSY couses.

Write out school names in Method (if using abbreviations).  Why is the % in each grade reported across sample when all analyses are reported by school?

says computerized for design- assume entire study was online
Did IRB at 2nd institution not review? line 241-242
Demographic measures lines 243-249 not complete sentences and dissemination info confusing here

Tell us how many myths you eliminated (line 253), how many news ones added. line 258 are the items from the textbook the original list or the ones you added (or both)
Validity of the findings: Reference to tables and figures is confusing. Primary data should not be in supplemental materials. And why is first table referred to Table 6?  line 296.  Vague references to Figures 1-3.

lines 309-316 talk about total scores- start there, then average scores.
If not showing full list of myths- should at least identify the 10 and 13 misconceptions- do these overlap or different items (line303-304)
pp 330-334- explain which way the difference went
Additional comments: Also APA style errors for in text citations- when use all authors or how to refer to a study in the wording of a sentence


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: MYTHS IN PSYCHOLOGY: PSYCHOLOGICAL MISCONCEPTIONS AMONG SPANISH PSYCHOLOGY STUDENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Dear Authors,

Thank you very much for giving me the opportunity to review the progress of the manuscript entitled "Myths in Psychology: Psychological misconceptions among Spanish psychology students".
It is very gratifying to see how both the feedback provided by the authors and the changes made to the new version of the manuscript, in my opinion, positively affect the quality of the review process and the resulting manuscript. For this I congratulate the authors.
With regard to the issues raised in my previous review, I consider that the authors have responded satisfactorily to each of them.
I understand that some of the issues I raise go beyond the stated objectives and I am pleased that this has been considered positively by the authors as a basis for further study.
For my part, I consider the manuscript to be of sufficient quality to be published in its current format and I consider it to be a relevant contribution to the literature.
Best regards,
Samuel P. León.
Experimental design: see "Basic reporting"
Validity of the findings: see "Basic reporting"
Additional comments: see "Basic reporting"


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: MAINTENANCE OF SPECIFICITY IN SYMPATRIC HOST-SPECIFIC FIG/WASP POLLINATION MUTUALISMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1.	Basic reporting: The manuscript is written quite clearly, in grammatical English; there are a couple of typographical errors.
Line 101: Correct to asympatrically
Line 357: Correct to where
All the elements of a professionally written paper are evident including the raw data. Figures and tables are also appropriate
Experimental design: 2.	Experimental Design: This study was designed to examine the possibility of hybridisation between two relatively closely related and sympatric fig species. If hybridisation does not occur, then what are the likely mechanisms of reproductive isolation between the species? The authors have conducted a host of appropriate experiments and observations to answer this question. The experimental design is therefore adequate. The methods are provided in adequate detail to allow for replication. The article is well within the journal's scope
Validity of the findings: 3.	Validity of the findings: The findings are well documented and the tests of the hypotheses have been adequately conducted. It is clear that there are reasons for reproductive isolation which the authors have clearly demonstrated.
Additional comments: In general, this is a well conducted study. I have a few suggestions about the ordering of some text which I have annotated in the accompanying pdf.


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

Input:
Title: MAINTENANCE OF SPECIFICITY IN SYMPATRIC HOST-SPECIFIC FIG/WASP POLLINATION MUTUALISMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This paper reports laboratory and field experiments on host-symbiont specificity in two Chinese fig-wasp mutualisms in which the host trees are members of a three-species complex. The data set is impressive and the authors are to be congratulated on the presentation of a comprehensive set of results ranging from the analysis of plant volatiles to behaviour of the pollinating wasps.

Overall, the work is fairly well-written and presented but suffers throughout via overly long text and frequent use of unnecessary jargon. I think the paper would be much improved by a sentence-by-sentence revision by a fluent English speaker. Additionally, the verbose use of language reduces clarity, especially in the introduction, meaning that it is not entirely clear why specific data were collected and why other data were not recorded (see below). Finally, it needs to be acknowledged that the paper will be read by a general reader and some text needs more detailed explanation. At the moment, I suspect that a non-specialist may not fully understand some text, why certain experiments and comparisons were made, and the interpretation of some of the results.

I think the data presented will certainly form the basis of a very interesting paper and fits the remit well for a general journal such as PeerJ. However, I think an extensive revision is required.

Specific comments

Why is the text not double spaced? Single spaced text makes things more difficult for reviewers, especially those who like to annotate a hard copy with red ink.

Abstract

The final sentence of the abstract is a good example of the use of excessive verbiage and jargon. Please simplify.

Line 39. What is a ‘diversity flux’?

Introduction

41-42 The first sentence is unclear. What unique components? Pollinator shifts between hosts seem to me to be more likely to promote hybridisation and gene flow rather than speciation.

51 Private code for what? This is vague.

51-59 Do you mean in different nursery pollination mutualisms there is evidence of host-symbiont coevolution and different host species produce their own VOCs especially attractive to their own pollinator species?

60 If there is strict one-to-one specificity, what causes the split? Allopatric speciation?

65 Is there any evidence of hybrid pollinating fig wasps?

86 There needs to be a less ambiguous description of the differences between monoecious and dioecious Ficus.

87 Receptive to what?

91 This is so only if 100% effective but nothing is perfect in nature, there is always variation.

92-95 A general reader is unlikely to fully understand this. More background detail is required.

103 Speculation or hypothesis?

121 If pollinated by ‘incorrect’ pollen, a fig may be more likely to abort, especially a female fig in a dioecious species. A male fig tree may still benefit by producing wasps that will disperse (hybrid) pollen. See Zhang et al. 2019. Ecology.

126 iii) Why was the reciprocal experiment not conducted using the opposite wasp/host combination? This needs to be explained as the question comes to mind several times when reading the paper. Also, why were seeds/galls not counted, nor fig abortion rates measured? High rates of abortion can exert strong selection on wasps to pollinate ‘correctly.’

M & M

134 Typo in latin name

134-136 Are the figs of the two species of similar size? I am aware that F. aur has very large figs, and there is a strong positive relationship between wasp and fig size among Ficus. If there is a size difference between the two systems, then due to correlated traits there will be a predictable difference in e.g. wasp head width, thorax width length, ovipositor length, all of which may affect the likelihood of a wasp entering a fig through the ostiole and thus the potential for hybridisation. All of this needs to be explained.

143 Debate? References needed. Do you mean ‘unclear’?

147 Traditionally? Or ‘usually’?

161 How were wasps identified? Characters?

162 How many figs for each tree per site? For data analysis, could a GLMM be used for e.g. wasp traits using site/tree as random factors to eliminate sampling error?

164 For figs of different stages were the same or different trees used?

170 Fig character differences? Pollen, flowers?

179 How were wasps collected/stored?

186 Detail on wasp and fig sizes required.

193 Between what?

196 How many wasps made no decision?

201 Sex not gender.

201 N Figs?

226 Why was only one ‘incorrect’ wasp / host Ficus combination tested and not the reciprocal combination? This needs to be explained.

240 Unclear to a non-specialist why this was done. More information on the natural history and general biology in the methods would help here.

246 How was randomisation achieved? Or was selection ‘haphazard’ rather than truly random.

249 More detail required on the measurements taken. Magnification? Was the ovipositor sheath removed? If wasp size measured it would need to be explained why, see above. I do not understand why fig traits were also not measured other than style length.

254 What is meant by ‘young figs’? What was the source of the wasps, and hence the source of the pollen? Seed counts, abortion rates (abortion needs to be clearly defined as this means different things to different people, especially botanists).

273 No mention of head width, thorax width before now so this just comes out of no-where. An explanation of fig size, wasp size and how these vary and may affect the direction of gene flow between trees and possibly wasps needs to be included.

Results

282 Contain offspring or foundresses? Please clarify.

316 Which VOCs were chosen to be investigated in more detail and why? This section is a little vague. Why were Ceratosolen sp. not tested?

327 What does ‘somewhat’ mean here? More clarity is needed regarding statistical significance (or not). Wasp size, fig size differences?

333. Own ‘proper’ host or ‘other, incorrect’ host? Please clarify.

335. Fig abortion seems really important here but no mention in the introduction or the methods. In the context of fig-wasp systems it needs to be defined and if possible, data presented as this could clearly act as a strong agent of selection in maintaining ‘correct’ pollination/oviposition behaviour in symbiont wasps.

337 This is interesting. More clarity is needed regarding differences own v incorrect host. Also interesting regarding size but more detail is needed. Ostiole sizes regarding a filter to wasps of the ‘correct’ size?

The results need to be clearer and more succinct throughout. I think if a small natural history section is added to the methods things would be a lot clearer especially to a non-specialist. If the introduction were made clearer and some specific predictions made, then I think it would be easier to produce a more succinct and clearer results section.

Discussion.

342 Imperfect host-symbiont alignment is probably very common and can be expected in an imperfect natural world. I think it would help to emphasise this point in the introduction and then go on to pose the question as to why we see such low rates of the ‘wrong’ symbiont associating with any particular host.

 351 Promoting v inhibition – vetoing does not appear to occur as there are always a few ‘mistakes.’

342-353 Please simplify the language and eliminate jargon wherever possible.

359 Natural rates of visitation were not recorded. Foundresses were counted but this is not the same as visitation because the wasps have visited a fig and then entered it. It would be most interesting to present visitation data though of correct v wrong wasp species on each of the single host fig species and how this does or does not differ to the patterns of foundresses within figs.

364. Still, not 100% precision but that is to be expected. We expect ‘mistakes’ and hence variation in such systems.

365 I am still puzzled as to why experiments using Ceratosolen sp. were not conducted.

373 Evidence for variation in body size per se and oviposition in agaonid fig wasps?

380 It would be interesting to know if the patterns of egg deposition, i.e how many eggs on average were laid by each foundress, varied according to wasp species and the species of fig it was ovipositing into.

381 In some Ficus with cryptic pollinator species, e.g. F. rubiginosa, different wasp species can reproduce in the same figs. If this happened in the species complex under scrutiny here, then the ‘wrong’ pollinator species may be able to successfully reproduce in the incorrect host if the host fig was pollinated and galled by another foundress of the correct species. E.g. the fig may be less likely to abort and the incorrect species wasp would ‘free-ride’ on the ‘correct’ species foundress. Of the figs collected, how many containing wasps of the ‘wrong’ species also contained a foundress of the ‘correct’ species?

389 Which sister species? Who are ‘they’?

419 References for this? Are there differences in abortion rates between males and females of the same species and do these vary according to which species of wasp entered and pollinated/oviposited? (see Zhang et al. 2019).

436 Seed data for all wasp-tree combinations?

438 Male figs should have lower abortion rates than female figs when they contain only the ‘wrong’ pollen.

448 Why not unaborted figs? What were the abortion rates?

454 This would have been best mentioned in the introduction.

465 Progeny fitness was not measured so this statement goes beyond what the data show.

472 Mechanisms presented show likely inhibition of gene flow between host species and thus against hybridisation. Which mechanisms presented will facilitate hybridisation? I do not think any convincingly show this here.

Figure 2 Is this figure necessary? If removed it would not detract from the story being told.

Table 1 Seed count data are interesting here but there is no mention anywhere in the paper of this. Also, reciprocal data are needed as are data for both host-wasp ‘correct’ combinations to act as natural controls.


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: MAINTENANCE OF SPECIFICITY IN SYMPATRIC HOST-SPECIFIC FIG/WASP POLLINATION MUTUALISMS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Fine
Experimental design: Fine
Validity of the findings: Fine
Additional comments: I enjoyed reading this interesting and thought-provoking paper. The topic of host specificity in mutualisms is of considerable ecological and evolutionary importance but there have been few studies of the consequences of atypical host use in specialist nursery pollinator mutualisms. I commend the authors on the use of a range of different methods in investigating this issue, including field surveys, characterisation of volatile attractants, behavioural responses and reproductive outcomes. Overall, this is a well-written and structured paper, reporting a well-executed study. My comments are mostly specific points requiring some clarification of details, or further consideration of arguments / explanations.

Terminology. I suggest the use of alternate host (as used in the conclusion) over non-typical host (as used elsewhere). I think this would be more transparent to the reader e.g. on line 19 in the Abstract. I am also not keen on the use of the word vetoing as in line 45. I think that alternate wording such as partner choice mechanisms or partner fidelity mechanisms would be clearer.

Line 46. Why did they have the potential to undermine extend biodiversity patterns? Surely the events themselves are part of the current biodiversity patterns that you document and underlying mechanisms help explain the existing patterns?

60. Strict specificity… leads to one-to-one interaction patterns. This is a circular argument.

82. Looks like a typo here. Should the sentence start “Ficus species…” ?

91. “thus ensuring..”. But they don’t, as you show in this study. “Promoting” is a better word here.

143. As the taxonomic status is still under the debate, can you confidently identify trees as belonging to one species or the other and how?

153. If I understand correctly, the wasps can only be identified to species under the microscope. If so, how did you get female wasps of known species to introduce to figs?

162. Did you collect ripe and immature figs at the same time such that they represent different cohorts, or were the ripe figs collected later and represent the same cohort of figs?  This distinction influences how you interpret the comparison of the situation in immature and mature figs?

278. It would be interesting to also test if the distribution of pollinators entering the alternate host fig is random across figs / trees.

333. It’s ery interesting to see that wasps produced more offspring (even if smaller) in the alternate host spcies. This could have significant evolutionary consequences and shows that that wasp size is strongly influenced by host environmental effect. The host shifting wasp becomes much more similar in body size to the normal pollinator of the alternate host. The sample size for these measurements is shown in Table 2 and is large, but it would also be interesting to know how variable it is across figs of the same species (and if correlated with fig size if you have data).

367-8 See query about fig cohorts above (line 162).

373 Is there here (or elsewhere) any evidence that smaller individual pollinator wasps have “compromised oviposition abilities”?

414 related to line 373 point. Be clearer about which host fig you are discussing here. Smaller size of C. emarginatus developing in wrong host (Fh) may decrease their success if entering the ancestral host (Fa) but not if they now seek out Fh, as these wasps are about the same size as the usual Fh pollinator.

442. Seem to be words missing here…

Figure 7. It would be informative to show the distribution of ovipositor lengths for both wasp species in each chart alongside the host fig style length distributions.


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: INCIDENCE OF HEMOPARASITIC INFECTIONS IN CATTLE FROM CENTRAL AND NORTHERN THAILAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: - English in the whole manuscript needs to be thoroughly edited.
- The scientific name should be written (italicized) in full when first cited in the abstract and in the introduction, then is abbreviated according to standard format.
Need correction in the whole manuscript
- Unify citation style, when citing multiple works (year of publication (the earliest first) or alphabetical order).
- Introduction:
    # Line 57-60: add a reference.
   # (Abdullah et al. 2019) didn’t investigate the effect of hemoparasites on animals health and the production of milk and meat.
Experimental design: -	Line 114: A thin smear of blood was collected?
-	Line 135: a graphic reading pad?  Do you mean a hematocrit reader card?
-	Some references in Table 1 are not related to the listed oligonucleotide sequences.
-	Unify the abbreviation for “microliter”. See line 127, 155, 163 and172.
-	Line 159-183: Add references when applicable.
Validity of the findings: -	Line 175-179: need a reference.
-	Line 228: If you calculate the multiple infection among each hemoparasite you will find that 100% (13/13), i.e., for all cases with B. bovis; 78.02% (9284/364) among A. marginale; 74.55% (290/389) among T. orientalis.
-	Line 310: Detection of this hemoparasite…..  which one?
Additional comments: References:
-	Order the reference list entries alphabetically.
-	Unify the style of the title according to the journal instructions.
-	Unify the style of the Journal name according to the journal instructions
-	Ref. 3: Article in Polish.
-	Italicize all scientific names.


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: INCIDENCE OF HEMOPARASITIC INFECTIONS IN CATTLE FROM CENTRAL AND NORTHERN THAILAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: This article is generally very clear and well written.
Experimental design: I only have minor comments related to experimental design:

1) The introduction discusses the global patterns of hemo-parasitic infections in cattle. It could be improved by refining the introduction to include specific information on what is known in Thailand. For instance, elaborate on the findings stated in lines 85-87.

2) The methods (lines 117-119) could be improved by elaborating on the specific morphology used to identify the parasites, to save on word count the authors could also improve these sections by including texts used to identify these parasites based on morphology.

3) The article would benefit by elaboration on how the phylogenetic trees were constructed. There are many methods that can be used to create phylogenetic trees (e.g., Neighbor-Joining, Bayesian inference, etc). The reproducibility of the methods could be improved by better outlining the methods in lines 193-198.

4) The statistical analyses (lines 200-204) are quite vague. Please specify the exact variable compared between the two groups (i.e, the packed cell volume)?

5) Lines 220-228: The population sample is really a subpopulation of the real population. Please specify standard errors around the prevalences using the sample size.

6) Would it benefit our epidemiological understanding of hemoparasite infections in Thailand by breaking down infection rates by region? I am curious if there are any meaningful spatial patterns in infection patterns.

7) The discussion refers to the proportion positive samples as incidence rate. These samples were not collected over time and hemoparasites can form chronic infections, therefore, it seems more accurate to refer to them as prevalence.
Validity of the findings: no comment
Additional comments: No additional comments