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

Input:
Title: GENOME-WIDE IDENTIFICATION AND ANALYSIS OF CYSTATIN FAMILY GENES IN SORGHUM (SORGHUM BICOLOR (L.) MOENCH)
Review round: 2
Reviewer: 4

Output:
Basic reporting: The manuscript entitled “Molecular characterization and expression profiling of cystatin family genes reveal their high evolutionary conservation and functional divergence in sorghum (Sorghum bicolor L.)”, summarized the systematic information about 18 sorghum cystatin genes (SbCys) and their expression and bioinformatics tools like protein motifs, chromosome localization, cis-elements.The authors focused on the cystatin genes and their possible regulatory mechanism that can provide the important information as well for the new researchers. please consider the following concerns and required to be addressed for brief description of the manuscript.

 1.	In introduction section, I would like to suggest that the authors need to enrich this section by describing the current status or response against biotic stress of cystein genes and phytohormones if latest literature is available.
2.	The regulation of gene expression and their responses to various stresses or with other species may also provide important clues and strengthen the author hypothesis please highlight in few sentences by providing fresh references in this section. If possible.

3. The author should clearly conclude the key results of the manuscript, how these genes are important and they can be beneficial/significance and what regard.

4. Please correct the grammar mistakes in the entire manuscript with a focus to the details of spelling, mistypings, italic and punctuation. There are small errors and enormous numbers of inaccuracy and negligence, please focus on too long or complicated sentences, which often describe the poor meaning.
Experimental design: Experimental design used in this manuscript is overall good. though, i have few comments for the author’s consideration hoping it will further improve this manuscript. Please fix the following comments.

3.	In M&M section, my particular concern is about the absence of significances and probabilities (P>…) for differences in the presented data. How many biological replicates (plants) were used for each of gene expression analysis?
4.	In line 189, Three independent  biological  repeats, The data collected three independent experiments” is unclear. If three plants were grown together and used as three biological replicates, this means one experiment. ‘Three independent experiments’ mean that plants were grown independently (simultaneously or consequently one-by-one) but number of used plants (biological replicates) must be indicated. Please clarify. Please also include ‘number of technical replicates’ for qPCRfor example ‘qPCR runs were repeated twice (or in triplicates)’. The phrase on L165 probably means the number of technical replicates but has to be clarified: “…from three independent PCR amplifications”. This will indicate for the accuracy of the used BioRad qPCR equipment and a Cycler Altogether, the received results with used number of biological and technical replicates have to be statistically treated with appropriate level of probability and significance.
5.	The M&M about abiotic stresses, ABA treatment is written absolutely incompletely, as a draft, where most of details are missed. This makes impossible to reproduce experiments by anybody, There are a lot of inaccuracies and negligence in this sub-section (as well as in entire manuscript) and MUST be fixed by Authors prior the submission. . Please insert sufficient information about hydroponics details, like how many plants/seedlings per container were used? How many containers in one experiment simultaneously or there were several experiments in time (see point 1)? Which water was used for hydroponics: tap-water, RO-water, distilled water, milli-Q water? Please provide either reference or full composition. The Growth solution was changed (how often?) or not? Aerated/non-aerated? for example: on the mesh, floating in holes of foam on the surface, in holes of the lid or another? All these and other sufficient details must be added in this sub-section.
6.	Following the comments please provide the description of how all stresses and hormones were applied to the plants/seedlings? Please make corrections accordingly. Firstly, ‘20% PEG 6000’ is not ‘drought’ but dehydration caused by absorption of water with PAG. Please replace it for ‘dehydration’ or ‘simulation of drought’. How PEG 6000 was applied to the hydroponics? What concentration, volume, quick/slow, single/several steps of adding to the hydroponics?
7.	Salinity is very different stress. Please indicate how young plants were transferred from regular Growth solution (without NaCl) to those with 250 mM NaCl? If so strong salt stress was applied to the young plants suddenly, in one step, it definitely will cause ‘osmotic shock’ and ‘cell plasmolysis’(see and use a recent Review: J Exp Bot 2013, 64, 119-127). How Authors manage their response with salt stress and salt shock?
8.	ABA is water insoluble but can be dissolved in ethanol and some other solvents. Please inserte your information in the text how ABA is dissolved and how applied to the plants? It was an adding of stock solutions with each of the hormones to the original Growth solution or a transferring of plants into a new Growth solution containing tested hormone?
9.	How much leaf tissue was used for RNA extraction, What volume and how pure RNA was received? How the quality of RNA was checked (for example, running on the gel) or not? Please indicate, if DNase treatment was included in the RNAiso TM Plus kit or not? Please provide more details about cDNA Synthesis kit (Promega, Madison, USA). for example, how much purified RNA was used for cDNA synthesis and how much cDNA volume was received in the end. Did Authors used non-diluted or diluted (indicate a number of folds) cDNA for qPCR analyses.
10.	Please clarify, whole flag leaf or which part of leaf were collected during sampling for RNA extraction, not well clear.
11.	My minor concern is that genes name should be mentioned rather using ID, the gene name may be given according to chromosome nomenclature.
12.	Did the author checked the other reference genes and their expression variation for qRT-PCR?
Validity of the findings: The findings of this manuscript is interesting and can be considered for publication. please improve the final manuscript with the following suggestions prior to submission.
Additional comments: No additional comments


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND ANALYSIS OF CYSTATIN FAMILY GENES IN SORGHUM (SORGHUM BICOLOR (L.) MOENCH)
Review round: 3
Reviewer: 1

Output:
Basic reporting: 1. Mention vegetative tissue not vegetable all through the MS.

2. Mention the classification and distribution of Cystatins in Introduction.

3. Cite all the latest references pertaining to this work. The author has not mentioned the Genome wide analysis of Cytstatin in Brachypodium (2017). (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5423411/). Mentioned in Discussion.
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: GENOME-WIDE IDENTIFICATION AND ANALYSIS OF CYSTATIN FAMILY GENES IN SORGHUM (SORGHUM BICOLOR (L.) MOENCH)
Review round: 3
Reviewer: 2

Output:
Basic reporting: In this manuscript, the authors have applied various functional annotations for 18 predicted sorghum cystatin genes (SbCys) using publicly available tools and data, rather than revealing cutting-edge facts with a meaningful perspective. Their approach is quite common and classical. Because of this reason, this manuscript seems to be interesting for only very limited researchers in sorghum field.  I strongly recommend to modify the story of the manuscript.
Experimental design: Although the methodologies do not contain any clear mistakes, the approaches were quite common and also old. Therefore, not so highly valuable information is presented in each figure, except Fig. 11.
Validity of the findings: In the manuscript, the authors only focus on 18 SbCys genes. Thus, the title “Genome-wide identification” may be a little inadequate. They do not show original genome-wide data, either. If this manuscript is focusing on the genome-wide functional annotation, the authors should show some original genome-wide data, such as ChIP-seq and RNA-seq data. If the authors focus on public data, then authors should take care of wider data in sorghum.
Additional comments: Please confirm the usage of following biological terms:
-	“Sorghum bicolor L.” should be followed by “Moench”?
-	Vegetable tissue -> vegetative tissue?
-	“infected” and “infested” were sometimes used incorrectly.


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND ANALYSIS OF CYSTATIN FAMILY GENES IN SORGHUM (SORGHUM BICOLOR (L.) MOENCH)
Review round: 3
Reviewer: 3

Output:
Basic reporting: The manuscript entitled “Genome-wide identification and analysis of cystatin family genes in Sorghum (Sorghum bicolor L.)” by Li, et al.; the authors performed a systematic study of the Sorghum cystatins (SbCys) gene family, and they also analyzed the tissue expression patterns as well as their response to abiotic stresses. The data will provide a foundation for further research of the better understanding the roles and regulatory mechanism of Sorghum cystatins in seed development and responses to different stress conditions.
Experimental design: I think the experimental design of this revised version meet the standards of Peer J.
Validity of the findings: no further comment
Additional comments: In this revised version, the manuscript is well-organized and clear to me. I do not have further suggestions on improving the quality of this manuscript.

Only some small gramma or typo issues

Page 1， Abstract， Line32, “one tested tissues” is not correct, please modify it

page 8：RNA samples were collected at 0, 12 and 24 hours, there should be a “,” between 12 and “and”; there’re still many such typos, please double check the whole manuscript, for example line 185 in the same page

Page  11， Line 262, “effect” should be “affect”?

Page 20 line 477, please re-phrase this sentence.

Should the transcriptome data be transformed to the server, so that you can get an accession number before it could be published?


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

Input:
Title: EVALUATION OF TWO FLUORESCENCE IMMUNOASSAYS FOR THE RAPID DETECTION OF SARS-COV-2 ANTIGEN—NEW TOOL TO DETECT INFECTIVE COVID-19 PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Dear Authors,

The word "immune assays", line 41, should be changed to immunoassays like shown on the title of your manuscript.

You could improve the sentence, lines 39 and 40, "However, PCR-independent techniques are urgently needed" by writing "However, rapid immunoassays for SARS-CoV-2 antigen could significantly reduced the COVID-19 testing burden currently weighing on laboratories around the world." This improvement can help an international audience clearly understand the message that you are trying to convey.

This is a minor change, but the sentence, lines 57, 58, and 59, should make clear that the COVID-19 disease did not become a pandemic until 2020. The SARS-CoV-2 virus was first detected in 2019.

Since you are evaluating two commercially available assays, you should add or compare your findings with the performance of each assay as reported by the companies (https://bestbion.com/wp-content/uploads/2020/05/bestbiondx-standardf-covid19-corona-ag.pdf and https://www.quidel.com/immunoassays/rapid-sars-tests/sofia-sars-antigen-fia). Your data appears to match their reported performances.

You considered cycles thresholds (Ct) values ≤40 positive. However, according to Sanguinetti et. al, "considering the current epidemiological scenario, a non-negligible proportion of symptomatic or, most commonly, asymptomatic patients, whose nasopharynx swab samples display Ct values of ≥25–<35 or ≥35, might be negative with STANDARD F COVID-19 Ag FIA (or similar) assays. This scenario would also encompass “new” patients who begin their SARS-CoV-2 infection course with a low viral load (resulting in Ct values of ≥35)." You should cite their paper (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7510559/).

You could address, in your introduction, the issue of PCR false positives that have been reported in many countries ("reinfections"). This could add to the importance of using rapid SARS-CoV-2 antigen tests (not as a replacement of PCR tests, but complementary). You can find some references here about this big issue:

https://www.cdc.go.kr/board/board.es?mid=a30402000000&bid=0030&act=view&list_no=367267&nPage=1

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7276353/#r2

https://ophrp.org/journal/view.php?doi=10.24171/j.phrp.2020.11.3.02

https://www.cidrap.umn.edu/news-perspective/2020/05/wha-passes-pandemic-probe-resolution-korea-clarifies-reinfection-reports

https://www.frontiersin.org/articles/10.3389/fcimb.2020.00445/full

https://www.cebm.net/covid-19/infectious-positive-pcr-test-result-covid-19/
Experimental design: Dear Authors,

Your experimental design could be improved by conducting viral cell culture tests along with RT-PCR. However, maybe you do not have access to a biosafety level 3 (BSL 3) laboratory to conduct viral cell culture experiments.
Validity of the findings: Dear Authors,

I think you should have cited previously published research (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7510559/) that used the same rapid assay(s) and addressed the same issue(s) as your manuscript did.

I think your results should have been validated using viral cell culture. Please use the paragraph bellow as a reference to improve your experimental design in the future.

"Conclusion Prospective routine testing of reference and culture specimens are necessary for each country involved in the pandemic to establish the usefulness and reliability of PCR for Covid-19 and its relation to patients factors. Infectivity is related to the date of onset of symptoms and cycle threshold level. A binary Yes/No approach to the interpretation RT-PCR unvalidated against viral culture will result in false positives with segregation of large numbers of people who are no longer infectious and hence not a threat to public health."

https://www.medrxiv.org/content/10.1101/2020.08.04.20167932v3
Additional comments: Dear Authors,

Your manuscript further validates the importance of using rapid antigen tests for COVID-19 as an important complementary diagnostic method. Rapid SARS-CoV-2 antigen tests could help safely reopen businesses, schools, and sporting events (https://www.nj.com/rutgersfootball/2020/10/big-ten-relies-on-daily-rapid-covid-19-antigen-testing-how-it-works-who-gets-tested-what-positive-tests-mean.html). Your manuscript is relevant to the current COVID-19 global pandemic and is in line with other publications related to COVID-19. This reviewer is conscious of the tremendous amount of work and effort that went into the manuscript. More researchers, like you, should focus on evaluating commercially available rapid antigen tests to help mitigate the global impact that the COVID-19 pandemic is having around the world by knowing the performance of those tests.


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: EVALUATION OF TWO FLUORESCENCE IMMUNOASSAYS FOR THE RAPID DETECTION OF SARS-COV-2 ANTIGEN—NEW TOOL TO DETECT INFECTIVE COVID-19 PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This work evaluated the performance of two commercially available antigen based point of care systems that can be potentially used in diagnosis of COVID-19.  Both systems are reported to have excellent sensitivity and specificity. The reported work is important but needs significant improvement before it can be considered for publication.
Experimental design: 1) The number of samples tested is low (only 64). If possible, I strongly recommend authors to include more samples so that the sensitivity and and specificity data reported will be more reliable.

2) It would be nice to include the details on calculation of specificity and sensitivity in the experimental section.

3) I suggest the information provided in line 113-117 along with more information provided in supporting information to put in a form of table.
Validity of the findings: 4) In a recent study, Liotti et al has reported the  performance of STANDARD F COVID-19 Ag FIA (SD Biosensor Inc.) with more than 300 nasopharynx samples (DOI:https://doi.org/10.1016/j.cmi.2020.09.030). On the basis of this work,  the authors should highlight the importance of their study; which I think is the comparison of analytical performance of two commercially available (and two country of origin) point of care systems.

5) The introduction section is poorly written. I recommend authors to briefly mention the analytical performance of other diagnostics methods (possibly in a separate paragraph) such as CRISPR, ELISA and antibody based methods. A nice summary on the performance of  these methods is nicely reported in a recent review [Anal Bioanal Chem. 2020 Sep 18 : 1–14.doi: 10.1007/s00216-020-02889-x ]

6) Line 128-135 can be moved to introduction section.

7) I recommend authors to include the simple working principle or schematics of the  (may be in form of figure)  assays method somewhere in  the introduction or discussion section.
Additional comments: 6) Please replace Covid-19 with COVID-19 in all places.
7) In line 41: replace immune assays with immunoassays


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: EVALUATION OF TWO FLUORESCENCE IMMUNOASSAYS FOR THE RAPID DETECTION OF SARS-COV-2 ANTIGEN—NEW TOOL TO DETECT INFECTIVE COVID-19 PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Dear Authors,

Thank you for making the changes that were suggested to improve your manuscript.
Experimental design: No comment
Validity of the findings: Dear Authors,

Thank you for citing the suggested previously published research that used the same rapid assay(s) and addressed the same issue(s) as your manuscript does.
Additional comments: Dear Authors,

Thank you for taking the time and putting the effort to make the suggested changes to your manuscript. Rapid testing is a big issue right now that should be addressed by more researchers like you.


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: EVALUATION OF TWO FLUORESCENCE IMMUNOASSAYS FOR THE RAPID DETECTION OF SARS-COV-2 ANTIGEN—NEW TOOL TO DETECT INFECTIVE COVID-19 PATIENTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: In the revised version, authors have have made/addressed the suggested changes.
Experimental design: In the revised version, authors have have made/addressed the suggested changes.
Validity of the findings: In the revised version, authors have have made/addressed the suggested changes.
Additional comments: Please check for English or any typos  (if any) very carefully.


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: GENDER-RELATED DISPARITIES IN THE FREQUENCIES OF PD-1 AND PD-L1 POSITIVE PERIPHERAL BLOOD T AND B LYMPHOCYTES IN PATIENTS WITH ALCOHOL-RELATED LIVER DISEASE: A SINGLE CENTER PILOT STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript entitled “Gender-related disparities in the frequencies of PD-1 and PD-L1 positive peripheral blood T and B lymphocytes in patients with alcohol-related liver disease - a single center pilot study” is an original research, well written (minor correction) and well-structured article that clearly highlights differences in PD1+ and PDL1+ T and B cells in patients with ALD, particularly focusing in a gender-based analysis. Literature references can be improved in the introduction.

Line 47 …well as PD-L1+ all T cell subsets in comparison with ALD males
Line 54 Define AH
Line 73 new diagnostic and therapeutic tools and reach new conclusions should be fostered
Line 74 In comparison to huge advances in the management of viral hepatitis (vaccines and oral therapies for HBV, oral regimes for HCV), alcohol-related liver disease (ALD) management has lagged.

Line 86 … that affect the function of multiple organs and tissues, potentially causing their failure

Line 96 ….PD-1 conducts signals only when IT is cross-linked with B- or

Lines 101-113 Contextualize the known evidence of PD1 in alcohol consumption and liver disease. Examples:

https://pubmed.ncbi.nlm.nih.gov/25479137/
https://pubmed.ncbi.nlm.nih.gov/30949049/
https://pubmed.ncbi.nlm.nih.gov/29935904/

Line 114 Since a great body of evidence indicates that compared with their male counterparts, women are more susceptible to the toxic effects of ethanol in the liver for any given dose of alcohol (Please reference this statement adequately) Like the reference stated in Line 234

Line 253 positive B cells, as well as PD-L1 positive T cells (all subsets) in comparison with ALD males

Table 1. Explain what is the p significance related to
283 CTP class B (Supplementary Table S8).
Experimental design: This works present original research through a methodological design that is adequate to determine the PD-1/PDL-1 expression on peripheral T and B lymphocytes, its correlation with markers of inflammation and the severity of liver dysfunction in the course of alcohol related liver disease (ALD) and with a gender focus that has revealed interesting differences.

Major comments:

Flow cytometry data regarding IMF and not only percentage of + cells should be analyzed and shown. Why are CD3+ cells (T-cell marker) shown when no triple staining (optimal) was done. Is it not enough showing CD4+ and CD8+ cells? If this marker is of some utility then it should be addressed in the analysis and discussion.
Validity of the findings: The results presented in this work are of a high impact to understand ALD. Nonetheless, it is important to address some of the following comments regarding how results are presented and discussed. I particularly urge the authors to improve how they explain the specific consequences of CD4+ and/or CD8+ cell dysfunction in the context of the results and the disease.  What should be expected if these cells have an abnormal immune response, how should this issue be addressed. In the case of B cells, serum Ig should be measured. What functional tests should be done to confirm that they have an abnormal immune response?

1.- Line 257 Analysis of mutual correlations between frequencies of PD-1/PD-L1 positive B and T lymphocyte subsets in patients with ALD.
In what table is this shown and what does this mean as it is not discussed.

2.- Line 264 Correlations of the frequencies of PD-1/PD-L1 positive T and B lymphocytes with conventional markers of inflammation in patients with ALD.
Why are CD3 and CD8+ cells for PD1 and PDL1 not shown?

3.- Line 266 Significant correlations of the CD19+PD-L1+ frequencies with all conventional markers of inflammation (i.e. white blood cell and neutrophil counts, C-reactive protein, and neutrophil-to-lymphocyte ratio) were found.
Include controls

4.- Line 284. "Moreover, ALD females with MELD >20 or mDF>32 differed significantly from ALD males with MELD>20 or mDF >32 in regard to the frequencies of PD-1 and PD-L1 positive lymphocytes (Table 5, 6, Supplementary Fig. S2, 3). "
Please describe what subsets were affected and which not

5.- I suggest merging tables for more integrated analysis, probably merging tables 3 and 4 and tables 5 and 6.

6.- “The association between the frequencies of PD-1/PD-L1 positive lymphocytes and
277 advanced liver dysfunction defined by MELD score >20 and CTP B and C class, and alcoholic hepatitis (AH) defined by mDF >32 was also checked. The significant differences were found in the female subgroup (Table 3, 4)”
No CTP B and C class are mentioned in those tables.

7.- Table 1 should show data from control subjects

8.- Why would only B cell PD-1 + lymphocytes increase in frequency? Can this differential increased expression between PD-1+ be explained?

9.- “Line 365 Our findings demonstrate the impairment in the regulation of immune suppressive mechanisms in women with the severe ALD course.”
There is no evidence of impairment, only the assumption based on markers. Functional tests should be performed.
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: GENDER-RELATED DISPARITIES IN THE FREQUENCIES OF PD-1 AND PD-L1 POSITIVE PERIPHERAL BLOOD T AND B LYMPHOCYTES IN PATIENTS WITH ALCOHOL-RELATED LIVER DISEASE: A SINGLE CENTER PILOT STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: In the present work, it suggests that there is a differential response in T and B cells in women with ALD by measuring the percentage of cells positive for PD1 and PDL-1. It was observed that in the general population there are no differences concerning healthy controls, however, when analyzed by gender, women with ALD presented significant differences for their healthy controls.
In this work, the expression of PD1 and PDL1 is defined as a marker of damage that can be associated with future complications such as neoplasms resulting from alcohol consumption. PDL-1 is associated with the presence of ascites, in addition to hypergammaglobulinemia in nonalcoholic liver diseases. PD1 and PDL-1 promote lymphocyte depletion by giving inefficient responses with a loss of functionality and antimicrobial activity.
In the background, it is necessary to delve into the immunological implications of expressing PD1 or PDL-1 in T and B cells, in addition to explaining that the female ones have important immunological differences to the male gender in the inflammatory diseases, so in this case, it is necessary to establish the objective of the work, it is not clear if PD1 or PDL1 is a marker of damage, a target molecule for a possible delay in liver damage, or the appearance of cancer, it is important to justify the relevance of this finding.
Experimental design: It is important to give clarity to the results, the way of presenting them is confusing, it is necessary to incorporate the data of the controls in all the tables, or to make a single table with the different conditions to be able to visualize the data and compare and conclude them.

Although the results are overwhelming in the female with ADL, the possible role of increased PD1 or PDL1 is not discussed, it is only mentioned that they are involved in unbalanced inflammatory processes. There is no direct immunological evidence that these lymphocytes are dysfunctional.

Immunoglobulins are not measured to conclude the hyperglobulinemia observed in other liver diseases. Table 7 is not clear the results can be presented in a better way. It is important in the discussion to mention the relevance of PD1 and PDL1 and its possible implication in pharmacological treatment or as a predictive marker of some neoplasm.
Validity of the findings: I consider it necessary to rethink the article and give it a more focused approach to the immunological implications of the PD and PDL1 expression in B cells in females with ADL, it is not clear what the objective of measuring it is. The article does not present any functional assay of these cells. There are limitations when working with samples, however, within the clinical history of the patients, the recurrence of infections or some other indicator that the immune response is altered would be very convenient.

I recommend presenting better tables or graphs that include controls, in addition to the fact that a better description of the test used in patients to determine ALD levels is useful, it is important to mention that they consist of the main text and not the supplementary material.
Additional comments: The results are conclusive, women with ADL increase the frequency of PD1 and PDL1 on their surfaces, however, it is necessary to justify the measurement of these markers in these liver diseases, to define what is the objective of this work, it is not clear if PD1 and PDL 1 will be used as possible markers of inflammation, complications or as a target for pharmacological treatment. The tables can be summarized and it is necessary to include the controls, although there is no difference. It would be useful to include some evidence of dysfunction in the immune system since the information is poor in biochemical and cellular markers. A medical history showing a higher incidence of infections or cancer may be helpful. The relevance of PD1 or PDL1 expression remains to be discussed, both molecules have different implications on the immune response. What is the function of PD1 and what is the function of PDL1? The response in women is different from that of men in conditions of inflammation, it is important to highlight these differences and their consequences on the body. In this case, what is the importance of expression in B cells?
Also, it is necessary to detail the selection criteria, the tests used as they were only mentioned but it is not described.


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 LARGE PTEROSAUR LIMB BONE FROM THE KAIPAROWITS FORMATION (LATE CAMPANIAN) OF GRAND STAIRCASE-ESCALANTE NATIONAL MONUMENT, UTAH, USA
Review round: 1
Reviewer: 1

Output:
Basic reporting: There are no fundamental problems with this paper. It is short and simple and there is little that is really debatable let alone problematic. There are however a fair number of points where the writing could be clarified and some additional small details or comparisons would make it rather easier to follow and better explain the author's position. All of the minor comments are presented on a marked-up document.

One thing that would help especially would be a simple figure (line drawing or photo) of the typical Cretaceous pterosaur radius and ulna to compare to the specimen.
Experimental design: This is all fine. Again, there are some areas that could be a little clearer.
Validity of the findings: Basically all fine. As above there are places where this can be more clearly expressed, especially in the lines about body size and overlap with other contemperaneous taxa.
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 LARGE PTEROSAUR LIMB BONE FROM THE KAIPAROWITS FORMATION (LATE CAMPANIAN) OF GRAND STAIRCASE-ESCALANTE NATIONAL MONUMENT, UTAH, USA
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this short communication, Dr. Farke presents a pterosaur ulna from the North American continental Kaiparowits Fm. Although incomplete and not that informative, the bone is worth publication because pterosaurs from this specific time interval (Campanian), environment (fluvial) and geography (western North America) are comparatively rare. The MS is written in plain, professional English, and I have just a single, very minor observation regarding the language. Dr. Farke’s work is self-contained, with all relevant data adequately provided within the text, figures and tables. The text is fully referenced, and the author was successful in providing an adequate background for his research.
Experimental design: As the studied specimen is not that informative, methods were restricted to morphological description and comparisons, what is adequate in this situation.
Validity of the findings: I agree with Dr. Farke’s identification of this bone as a right ulna and with the provided size estimations. Still, I believe that the author’s conclusions are too unpretentious and that this specimen welcomes a small piece of (well-grounded) speculation regarding the owner of the bone. In my understanding, pterosaurs from the Campanian are restricted to pteranodontids, nyctosaurids and azhdarchids. It is very unlikely that a nyctosaurid produced this bone: not only nyctosaurids were comparatively small animals, but they also had much more slender ulnae. The ulna of Pteranodon seems slightly less stout than RAM 22574, but pteranodontids with similar ulnar proportions do exist (e.g. Longrich et al., fig. 4). More interestingly, pteranodontids are indeed strongly correlated to coastal/marine environments (I know no exceptions). Accordingly, the size, faunistic context (azhdarchid remains were found in close association to the new specimen), and lithology (azhdarchids are more likely than not found in continental deposits) makes an indeterminate azhdarchid the most likely owner of this particular bone. Although speculative, this is apparently the most parsimonious hypothesis, and the author may want to acknowledge this in his text.
Additional comments: Geological Setting: Please indicate more clearly the depositional environment of the Kaiparowits Fm.

Lines 15-16: Azhdarchids seem to challenge this particular bias, with most records coming from inland deposits (see Witton and Naish, 2008).

Line 124: femora are also much more slender and slightly sigmoid, very different from what is displayed by RAM 22574.

Line 144: delete 'in'


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 LARGE PTEROSAUR LIMB BONE FROM THE KAIPAROWITS FORMATION (LATE CAMPANIAN) OF GRAND STAIRCASE-ESCALANTE NATIONAL MONUMENT, UTAH, USA
Review round: 1
Reviewer: 3

Output:
Basic reporting: The contribution is well written, the references are up to date and sample classic and recent publications, the hypothesis of identification is clear and the discussion/conclusions follow the data (the study is repeatable based on what is presented). The text is clean minus a typo here and there (see pdf)
Experimental design: The paper absolutely falls within the scope of the journal. All permits are cited and the methodology is detailed. It would be nice if the author provided a 3d model of the ulna if possible.
Validity of the findings: The conclusions are based on sound data and the author indicates where there could be interpretation differences. The implications are provided and are backed up by data.
Additional comments: This paper is nearly ready to go. Other than a few tiny items on the pdf, my biggest suggestion is to add a map and maybe a strat section that summarizes the geologic and geographic occurrence.


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 LARGE PTEROSAUR LIMB BONE FROM THE KAIPAROWITS FORMATION (LATE CAMPANIAN) OF GRAND STAIRCASE-ESCALANTE NATIONAL MONUMENT, UTAH, USA
Review round: 2
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: I'm happy with the final version of the MS.


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 LARGE PTEROSAUR LIMB BONE FROM THE KAIPAROWITS FORMATION (LATE CAMPANIAN) OF GRAND STAIRCASE-ESCALANTE NATIONAL MONUMENT, UTAH, USA
Review round: 2
Reviewer: 2

Output:
Basic reporting: This paper is ready to go with a few additional suggestions based on changes from the first draft. I think the author did an excellent job incorporating reviewer comments.
Experimental design: NA
Validity of the findings: Excellent as was before.
Additional comments: Minor comments:
Systematic Paleontology
-it would be helpful to provide the phylogenetic definition – e.g., Archosauria sensu Gauthier and Padian 1985
-I would like to reiterate you do not need the Pterosauria Genus and species indeterminate.
 	-it is redundant given that every individual belongs to a genus an a species
	-it is also a vestige of Linnean taxonomy – I would be happy to chat about it if needs be

Line 254 – ‘truly gigantic pterosaurs’ please give a wing span range to slightly quantify

Figure 2 – the head of your large size of Cryodrakon is cut off, move down or over a bit?
-nit picky on my part, but I think your outline of the coasts can be less rounded (maybe create in a non-vector program) so that it matches the real coastline better.
	-make the boundaries of the US (gray) have less weight because it clearly overlies the coast line and enters bodies of water
	-given that you have political boundaries, it is necessary to have countries labeled for non-US researchers.

I am happy for this to be published without any more reviews from me


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

Input:
Title: THE DIVERSE RADIODONT FAUNA FROM THE MARJUM FORMATION OF UTAH, USA (CAMBRIAN: DRUMIAN)
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Pates et al. (2020) provides an exceptional description of new taxa that greatly increase the known diversity of Laurentian radiodonts. I enthusiastically support the acceptance of this paper and am looking forward to it being shared with the greater community. The only thing that I would suggest is to include stratigraphic sections to drive home the facies inference side of the story; however, I see that many of the specimens included cannot be tied to particular sections (let alone particular localities) so the point may be moot.


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

Input:
Title: THE DIVERSE RADIODONT FAUNA FROM THE MARJUM FORMATION OF UTAH, USA (CAMBRIAN: DRUMIAN)
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: Results are supported by evidence but it would be wise to concede that the interpretation of ecology is somewhat speculative.
Additional comments: This is an interesting and important account of the radiodont fauna of the Marjum Formation, one of the major Utah Cambrian Lagerstätten. It includes the description of what is convincingly a new taxon, Buccaspinea, and new reports and information on other radiodonts from the formation. The paper is well written and illustrated. There is a useful comparison of the major soft-bodied Utah assemblages, supported by faunal lists (Supplement) and an extensive discussion of the ecology of the radiodonts which is somewhat speculative (and for that reason I would not lead the title with ecology).

The text and illustrations could be condensed if necessary.

I offer the following suggestions for improving the manuscript (numbers refer to lines).

19 I wouldn’t describe them as common – as top predators, for example, they tend to be rare compared to other elements of the fauna.  If you mean radiodont taxa are commonly present that is something different.

20 Seilacher’s concept of Konservat-Lagerstätten extended to well-preserved skeletons.  Your definition is too narrow and therefore incorrect.  What do you mean by ‘prominence’?

39 You have no evidence for ‘high relative abundance’. A single specimen of Buccaspinea tells you nothing about how rare or common the animal may have been.

50 Konservat Lagerstätten do not equate to strata.

53 What do you mean by ‘equally prolific’ – is diversity similar, fossils similarly abundant …?

81 ‘the latter’ (not these) to make it clear that you are referring to ‘anomalocaridids’.

86 I wonder if the two rami in a paired flap could function indepently to the extent of one contributing propulsion while the other was used for stability and steering.

95 ‘three to five’ presumably out of seven BS genera. Why can’t you be more precise?

113, 147 There’s no merit in giving two estimates for diversity – one more up-to-date than the other.

120 Or it presumably might simply be taphonomic.

149 ‘may be partially explained’ So how does this compare to the stratigraphic spread/duration of fossil occurrences in the other formations?

228 These remarks as a whole refer to genera of Hurdiidae. The first sentence, however, refers specifically to Buccaspinea – that part of the discussion should follow the heading on line 261.

308 ‘are visible from the head region’ is an odd way of indicating that they are the only parts of the head region preserved.

321 ‘The boundaries between the plates in the oral cone are not clear’, but nor are those in the anterior appendage - at least in the illustration. The holotype of Buccaspinea is generously illustrated but what is lacking is a close up view of the anterior appendages in low-angle illumination that would complement the drawing and illustrate some of the detail. Only the overall outline of the appendages is evident in the illustrations.

516 Given the preservation of new morphological details it would be useful to have a reconstruction of the outline of the carapace of Pahvantia.

560 ‘a close phylogenetic relationship’ not relationships.

750 ‘co-occurred in the same ecosystem’ But is there any evidence that they actually co-occurred? Presumably they could have lived in different places and at different times.

756 This section needs to be prefaced by some acknowledgement that the sample size is very small, the animals are not readily preserved, etc. The text could be reduced with reference to Table 2.  You don’t have the basis to do more than speculate about ‘the local evolution of radiodont faunas in both space and time’ – you’d need abundance data etc.

850 Here and elsewhere it’s not clear why bivalved arthropods is in parentheses, i.e. ‘bivalved arthropods’. These animals were clearly bivalved and there is nothing to suggest an affinity other than with arthropods.

902 If RAR provided data on stratigraphic positions within the Marjum, why is that information not included – or have I missed it?

Figures 2-6 Every figure legend does not need to repeat all the abbreviations of morphological details.

Figure 6: The photographs do not show the evidence for the morphology of the limbs. You should add a low angle illustration of the anterior appendage to show the breaks of slope that would distinguish the endites, likewise to show that the three cusps of the teeth in the oral cone are not at different levels (i.e. belong to a single tooth).  Neither the attitude nor robust nature of the auxialiary spines on the endites are matched in the reconstruction.

Figure 11.  Why ‘putative’ hydrozoan?  Why ‘bivalved arthropod’ in parentheses? If you don’t agree with these interpretations you should justify your view or provide a citation.


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

Input:
Title: THE DIVERSE RADIODONT FAUNA FROM THE MARJUM FORMATION OF UTAH, USA (CAMBRIAN: DRUMIAN)
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is an excellent contribution, scientific important, well-done in all respects, and well-written. The language is clear and unambiguous. I think this will stand as a landmark paper in our developing understanding of this group of arthropods.

I have little to add to the paper, or to change, other than some seminal papers or useful reviews that were overlooked or not deemed central to the main points of the paper. These relate to global sea level (which becomes an important part of the argument), biogeographic/paleogeographic patterns, and to agnostoid/'polymeroid trilobite' life habits and classification. The phylogenetic relationships are still in discussion, and citing some of the reviews would be useful.
Experimental design: Excellent. This work is replete with new information, and updates of older data, on a taxonomic group of great importance for understanding the dynamics of the Cambrian evolutionary radiation and the ecology of this group. The research questions are clear, and well researched, with lots of detail provided.

My only concern is with the numerical calculation of swimming vs. non-swimming Cambrian animals. The methodology is not indicated, and it should be. 'Polymerid' trilobites, if they are included as benthic, may be skewing the interpretation toward in the direction of benthic animals. This is unlikely to be true. The trilobites are diverse, and varied ecologic niches are likely. Many were probably nektobenthic. (This is not a major point of the paper, but it is worth a second look, and a clarification of how the information was obtained.)
Validity of the findings: Basically excellent in all respects. My only concern is with the numerical calculation of swimming vs. non-swimming Cambrian animals (noted above). The information relating directly to the radiodont fossils is quite good.
Additional comments: I added comments directly on the ms Word file, and added some key references that should be cited. Please see them.

With quite minor revision, this article is certainly acceptable for publication, and I do not need to see another version.


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

Input:
Title: THE DIVERSE RADIODONT FAUNA FROM THE MARJUM FORMATION OF UTAH, USA (CAMBRIAN: DRUMIAN)
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors have gone above and beyond to address reviewer concerns. I am still very excited to see this research be made available to all, especially to those of us who pursue questions related to the diversity/disparity of stem group arthropods and the evolutionary dynamics of Arthropoda as a whole.


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

Input:
Title: THE DIVERSE RADIODONT FAUNA FROM THE MARJUM FORMATION OF UTAH, USA (CAMBRIAN: DRUMIAN)
Review round: 2
Reviewer: 2

Output:
Basic reporting: This revised version takes full advantage of the comments of Reviewers 2 and 3 (Reviewer 1 merely provides an endorsement) and should now go forward for publication.  I only offer very minor comments for further consideration.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: You provide a robust rejection of the possibility that differences between the Konservat-Lagerstätten might be taphonomic in response to my comment on the last paragraph of the Introduction.  You might consider including these observations in the paper.

Retain the comment on Buccaspinea at the beginning of Remarks (under Systematic Paleontology) if you wish, but name Buccaspinea rather than calling it ‘this new taxon’

On line 879 you mean ‘sequence’ not ‘sequential’.


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

Input:
Title: IDENTIFICATION OF HUB GENES AND BIOLOGICAL PATHWAYS IN HEPATOCELLULAR CARCINOMA BY INTEGRATED BIOINFORMATICS ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. Several figures, such as Figure1 and Figure7, was unclear to understanding.
2. Typing errors or ambiguous words, like line 164 “functionally significant”?
Experimental design: Methods and data resources lacks detailed information.
1. What does adjacent tissues mean? Why you define them as adjacent samples?
2. What is the cutoff of a gene with lower expression? And why you choose the rule?
3.The values of PCC (Pearson correlation coefficient) can be -1 to 1. I think the 0.65 cutoff is roughly without any reference or statistical explanation.
4. The description of your association matrix was too simple. How to build it and what’s the calculation formula?
5. The method of hierarchical clustering is unclear, is it average correlation value or others?
6. The legend of Figure 2 was rong. It should be log2(Fold
Change)=±1
7.In Figure 3, there are several networks. The author only used the largest one. They did not discuss why they are smaller and the possible function of the smaller networks.
8. The details of modules were unclear. How many genes were included in the 9 modules? And why to remove modules with genes less than 50?
9. In Line 158, the two BP entries were not found in figure 6.
10. The heatmap of Figure 10 should be sorted or clustered.
Validity of the findings: 1. Due to lack of data resources, it is uncertain if the test samples(GSE138485) were involved in the tissues samples of network construction.
2. In the validation section of 13 hub genes, several genes are up-regulated and down-regulated together in the same cancer, why?
3. The Receiver Operating Characteristic Curve (ROC) and AUC value are suggest to validate the function of 11 hub genes.
4. The analysis of DNA methylation is too simple. There are several kinds of DNA methylation，so which methylation was analyzed here?
5. In line 274-276, why you mentioned the lncRNA, is it correlated with your results?
6. In the discussion section, there is too more introduction of GO entries, and lacks validations to confirm the hypothesis that those hub genes are biomarkers of HCC.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF HUB GENES AND BIOLOGICAL PATHWAYS IN HEPATOCELLULAR CARCINOMA BY INTEGRATED BIOINFORMATICS ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English language should be improved. Some examples include lines 82, 149, 165, etc.
In lines 82 and 149, “a association matrix” should be “an association matrix”.
In lines 165, “9 topological analysis algorithms” should be “Nine topological analysis algorithms”; it is better to use English words instead of Arabic numerals less than ten.

The logic of the introduction needs to be improved. The introductory part should provide an analytical framework for the current research. Relevant prior literature should be appropriately referenced.

In lines 57-59, the details of the dataset(s) and samples should be provided.
In line 66, P-value ≤ 0.05 was used as significant, while in other parts of the manuscript P<0.05 was chosen. It should be better to unify the standard.
In lines 100-104, the details of the 9 topology analysis algorithms should be provided.
In Figure 2, it seems that down-regulated (n=471) and up-regulated genes (n=4,130) were reversed; Additionally, bar colors seem to be pink and green, not red and blue.

The results and discussions are not sufficient. Need to discuss the causality or relationship of modules, gene functions, pathways and HCC stages.
Experimental design: no comment
Validity of the findings: Impact and novelty not assessed.

The data in the results section should be displayed more to ensure that the results are credible rather than statistically false positives. For example, in lines 133-137 & 142-143, since there are so many genes and interactions, does the screening criterion only depend on the size of the subnet? As far as we know, the greater the amount of data, the greater the power (detection effectiveness); In addition, the occurrence of cancer may just be some mutation(s) of a single gene or genes related to its pathway, so small subnets with special functions also need attention. Another example, in subsection 3.3 (lines 147-161), the module m1 has a high risk of false positive. It is hard to explain why m1 was associated with meiosis. Since the occurrence and progression of tumors is a multi-step process, a more reasonable approach is to subdivide the modules and correspond to different stages. Supplementary materials need to add necessary notes.

In the present study, the analysis framework seems not clear enough. The purpose and function of each step of analysis need to be clearer. For example, in lines 59-60, why genes with lower expression should be removed? In lines 82-86, why the clinical indicators were divided into event, T, N and M stages? Moreover, Pearson correlation analysis is too simple for the association analysis between gene modules and clinical indicators. Time series analysis model, such as survival analysis should be conducted. In lines 99-105, what is the basis or reference for “confidence score ≥ 0.400 was set as significant” or “5 genes with the highest score in each algorithm were merged together as the hub genes”? In lines 114-115, parameter settings for random forest(s) need to be provided.

Conclusions are not well stated.
Additional comments: Sufficient research should be done before starting the study. Previous literature(s) related to this study should be cited and the results should be compared with the present study.


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

Input:
Title: IDENTIFICATION OF HUB GENES AND BIOLOGICAL PATHWAYS IN HEPATOCELLULAR CARCINOMA BY INTEGRATED BIOINFORMATICS ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment.
Experimental design: 1. In figure3, the total number of gene and the number of gene in GCN should be listed. How much gene were filtered based on the smaller networks?

2. The author finally identified 9 modules. How to define a module is “too few/outliers” and a module is “too large”?  Is there any detailed parameters ?

3. In the validation section of 13 hub genes, author explained several genes up-regulated and down-regulated together were due to inconsistent data. Thus, what is the problem make them inconsistent? Did you make quality control before data processing? Are these data sets credible?

4. There are three types of DNA methylation, CHH, CHG and CG. Which kind did you analyze here?
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: IDENTIFICATION OF HUB GENES AND BIOLOGICAL PATHWAYS IN HEPATOCELLULAR CARCINOMA BY INTEGRATED BIOINFORMATICS ANALYSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Supplementary tables need to be sorted logically, for example, in ascending order of P value. In addition, some headers need to be annotated.
There is still some spelling or grammar errors, such as "mutilevel" in line 198.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Regarding the meiosis in the m1 module, it is better to provide some explanation in the discussion section. There are several articles on meiosis and tumors.


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: KNOWLEDGE AND ATTITUDES TOWARDS STEM CELLS AND THE SIGNIFICANCE OF THEIR MEDICAL APPLICATION AMONG HEALTHCARE SCIENCES STUDENTS OF JOUF UNIVERSITY
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this manuscript, the authors aim to assess the knowledge and attitude of the medical and dental students towards the stem cells and their applications in medical science. The manuscript is clearly written and it is dealing with an interesting topic.
The authors may consider the following comments:
Line 120: please, make sure that the reference Browne 1995 ("On the use of a pilot sample for sample size determination") is adequate to explain the knowledge and awareness among public and healthcare staff about CB banking.
Section Statistical Analysis: in the description of the statistical methods, add the ANOVA test and the independent t-test that you used for comparison of the mean values of knowledge and attitude scores according to socio-demographic characteristics of the sample, as indicate in Table 3.
Lines 187 and 217: Percentage of married students is reported as 7.2, while in Table 1 as 7.4. Please, resolve this inconsistency.
Line 208: it's unclear the meaning of the words "do. Published 2020". Please, clarify the meaning. If it refers to a work unpublished, cite it as "unpublished data" and supply the author's first initial and surname, and the year of the data collection, as indicated in the "Instructions for the authors".
Lines 238, 252, 266, 274: please complete the reference "AlAbdulqader et al." with publication year.
Line 301: complete the reference with publication year, full title of the Journal, volume: page extents.
Line 319: it's unclear the reference "do. ScWtaawt. Published 2020". Please, clarify.


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: KNOWLEDGE AND ATTITUDES TOWARDS STEM CELLS AND THE SIGNIFICANCE OF THEIR MEDICAL APPLICATION AMONG HEALTHCARE SCIENCES STUDENTS OF JOUF UNIVERSITY
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This is an interesting study and the authors have collected a unique dataset using cutting
edge methodology. The paper is generally well written and structured. However, in my opinion the
paper has some shortcomings in regards to some data analyses and text, and I feel this unique dataset
has not been utilized to its full extent. Below I have provided numerous remarks on the text as it is
often vague and long-winded

In the abstract:
You need to avoid "We" and try to use passive voice.
You need to delete some of the methods part and rewrite it for example no need to mention the statistical analysis part in the abstract.
The abstract was copied from the text of the article, which needs some modification.

Methodology section:
In the methodology and sample size calculation, the formula was used contain "d2", the margin of error e2 and also the authors need to write the name of this formula "cochrane". and referred p to proportion will be better than prevalence.
The authors, need to mention the sample size method they use it (i.e. stratified or random,..etc)
In the ethical part "Jouf university" the first letter should be capital.
In the statistical analysis section:
Noting mentioned about hypothetical test such as ANOVA and independent t test but both of them presented in table3.

In the result section:
No need for figure 1, it can be added to the table 1.
No need for table 4, the sentence already mentioned in the text is enough.

The authors need to do some more analysis to assess the awareness of the participant about Stem Cell Donor Registry and their year of study or gender (cross tab and Chi-square), also to add chi square to analysis section of the methodology.

Discussion:
Nothing mentioned about the limitation of the study, the authors need to mention some limitations to be avoided in the future studies, and in some part I feel like it is a rewritten of the result section.

In line number 229, the authors mentioned "Majority of the other studies.." and only one reference was used, what about the other studies!
The authors compare their results with other studies were done in KSA, why no international study was mentioned?

Conclusion:
Need to rewrite to avoid active statement "We" and use passive voice. Also, some parts of the conclusion was written like a result not conclusion i.e. p- value, these sentences should be deleted.


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: KNOWLEDGE AND ATTITUDES TOWARDS STEM CELLS AND THE SIGNIFICANCE OF THEIR MEDICAL APPLICATION AMONG HEALTHCARE SCIENCES STUDENTS OF JOUF UNIVERSITY
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors responded satisfactorily to my comments.


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

Input:
Title: KNOWLEDGE AND ATTITUDES TOWARDS STEM CELLS AND THE SIGNIFICANCE OF THEIR MEDICAL APPLICATION AMONG HEALTHCARE SCIENCES STUDENTS OF JOUF UNIVERSITY
Review round: 2
Reviewer: 2

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: I want to thank the authors for their response to my comments, most of my comments taken in consideration, I want to add:

In cronbach alpha as the authors calculated from pilot study to be more than 0.7, it is better to add a comment like "which exhibited  good internal consistency in the format used" or any other comment.

The authors calculated the sample size two times, one is enough.

The authors used skewness and kurtosis to test the normality, using, Kolmogorov-Smirnov or Shapiro are better depends on the sample size in each group.

The authors wrote they used ANOVA to test the association..., for association Chi square test is used, ANOVA used to compare between group as a hypothetical test.

Instead of "proforma" word, instrument is better and more usable, same with "question" statement or item is better.

In table 3, attitude section Mean+-SD not mean (SD),year of education 4th should add +-, smoking status for attitude need -, under table 4 please write the test used and statistically significant at which level 0.01 or 0.05 for example. Please review the tables for any error.

Other than that, I don't have any more comment and I want to than the authors again.


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

Input:
Title: KNOWLEDGE AND ATTITUDES TOWARDS STEM CELLS AND THE SIGNIFICANCE OF THEIR MEDICAL APPLICATION AMONG HEALTHCARE SCIENCES STUDENTS OF JOUF UNIVERSITY
Review round: 3
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: I want to thank the authors for their responses to my comments, the manuscript is very well written; clear, precise, and easy to understand. For me,  all comments were taken in consideration and now it is ACCEPTABLE.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Abstract:
External environment should be replaced by ‘adverse environment’
Keywords were missing
Line 44, remove ‘while’
Introduction
1.	The ATP-binding cassette subfamily G is the largest subfamily of ABC transporters family. It was found that ABCG protein was widely distributed in plants and participated in many key physiological processes. Need reference.
2.	Line 59, ‘including’ should be replaced by ‘includes’
3.	Line 61, ‘with’ should be added before ‘approximately 120 amino’
4.	Line 83, it – It
5.	Line 80-83, 85-86 reference
6.	Line 91-98 mainly described the result you have obtained, which is not suitable to put in the section of Introduction. Please add your hypothesis of the study.
M & M
1.	The seeds were sown and after 1 year, the samples were taken. The greenhouse information you provided is far more than enough since the paper only mention the photoperiod of 16h/25°C day and 8h/16°C night. This is not possible that during whole year, the temperature is so stable and the light length as well. Another question is the light level, humidity and irrigation system.
2.	‘One years’ to be replaced by ‘one year’
3.	‘A incubators’ to be replaced by ‘incubators’
4. why did you choose 6 h and 12 h after stress?
5. Why is it 1 year after growth that you took the sample
Discussion
1.	Was the ABCG the largest group in rice and Arabidopsis?
2.	Fig 2 was not discussed indicating it is less important?
3.	In our study, CcPDRs may be involved in the response of pigeon pea to abiotic stresses such as drought, chilling, salt, while CcWBCs had the ability to deal with high temperature, drought and aluminum stress as show in Fig.5B. I am confused about which belongs to CcPDRs and which was CcWBCs in Fig. 5B.
4.	Line 317-321. Long sentence that need to divide. Moreover, it need reference to support. The same for line 323-328.
5.	Line 329-346 need more and deeper discussion. For instance, any proof from previous studies indicated that the expression of the ABC in different organs were diverse. The up-regulation of the genes at abiotic stresses corresponded to the previous studies or not and why?
6.	The discussion need to deeply discussed by combining with previous publications to improve the depth of the paper.
Conclusion
The results were missed since it only mentioned what you did.
Reference
     Make the format corresponding to the requirement from the PeerJ.
Experimental design: The experimental design are ok even though there were something remaining unclear.
Validity of the findings: The results were validated by qRT-PCR. Even though this validation is fundamental, the author did show some proof from the result of qRT-PCR.
Additional comments: No additional comments


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: 1. The authors should work on the language. The entire article suffers from this one way or another.

2. line 93:ABCG transporters...distributed the cell membrane. What does it mean?

3. line 221: The authors mentioned that they performed subcellular localization. It should be termed as a prediction of subcellular localization.

4. line 223: The authors should clarify the CcABCGs are predicted to be localized in the cell membrane.

5. line 225: The statement that the ATP produced in photosynthesis provides energy for ABCG transport is not supported by any fact. Just because something is predicted to be localized in the chloroplast doesn't mean a direct correlation between the ATP usage and transport through the channel.

6. line 238-242: Additional information regarding the homology modeling is required. What was the starting model? How much sequence/structural similarity between the source and the query? Also, what do we learn from this? The authors mentioned only the number of TMD and NBD domains in the proteins as an outcome of the modeling experiments. Further analysis of the models could shed light on the structure-function relationship of these proteins and may make the manuscript significantly stronger.

7. line 243-250: The authors reported the analysis of the exon/intron composition of the genes. What is the significance of this with respect to the paper? How this is related to the abiotic stress response of these gene-products?

8. line 257-259: The observation of the presence of the low-temperature related elements should be investigated in detail. Are the sequences of these elements very conserved? How about between different plants?

9. line 273-274: The word almost significant could be replaced with proper statistical numbers and significance.

10. line 317: The authors stated that the promoter analysis were done in Fig. 3. There are no promoter analysis data reported in Fig. 3.

11. Each and every figure legends should be rewritten. Figure legends should be self-sufficient and must contain all information to decipher the figure.

12. Figure 2: The location of the genes in the respective chromosomes were shown meticulously. However, how is this information central to the paper? This should be detailed in the text.

13. Figure 3A: Are the sequences of the specific domains really necessary to be inside the figure? Are these sequences absolute or just consensus? If they are consensus, how were they obtained? How much differences are there between the domains in different proteins?

14. Figure 3B: Macromolecule structures visually look better with a white background. I am still not clear what are these two models telling us about this story? Figure legends should be more descriptive.

15. Figure 4A: What this have to do in the paper? Does this figure contribute in any way towards validating the findings/main story? Please clarify.

16. Figure 5B: What is CK? Error bars have been used -- are they SD or SEM? How were they calculated? Needs to be mentioned in the text.

17. Figure 5B: Are the differences that are seen here significant? Simple statistical analysis should be done on the numbers and always reported with this type of experiment.

18. This article needs a proper introduction to various types of abiotic stresses and their importance. Why used AlCl3? Why is that a key chemical deserved to be tested? Pertinent information like this is a prerequisite for a complete story.

The authors have done a significant amount of work for this article. However, the results were ineptly written and presented. In my view, the authors tried to report many information that are not related directly to the story and thereby diluting the core message. Also, the parts which could have made the manuscript stronger were unfortunately either poorly analyzed or negligently reported.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors present a bioinformatics and expression analysis of the ABCG transporters in pigeon pea (Cajanus cajan).
1. The writing and organization of the paper MUST be improved.
line 42: ABC transporters plays…?
line 47: was that the AtABCB1 (also known as AtMDR1) was cloned in Arabidopsis in 1992? line 56: According to their structure…?
Line 56: ABCG subfamily were…?
Line 58: while PDRs belongs to…?
Line 60: The NBDs domain consists of…?
Line 64: the hydrolyzes reaction of …?
Line 68: more than 40 ABCG transporters members…?
Line 73: “the plant's tolerance salt and drought stress…” should be “the plant's tolerance (to) salt and drought stress…”
Line 76-79, line 87-90…
Line 92-93 ABCG transporters in pigeon pea was located on…?
Line 94: classed into..?
Line 96: stress of temperature, drought and high concentration of aluminum…?
…. …. …. ….
I can scarcely read it. Simple grammatical errors can be seen everywhere.

2. In motifs analysis, the authors were identified 10 conserved motifs in the CcABCGs, and these conserved motifs were annotated by InterPro. However, no conserved motifs were annotated as NBD or TMD domains, which were the typical features of ABCG subfamily members.

3. According to the title “ABCG transporters in pigeon pea”, the authors should focus on ABCG transporters in the manuscript. First, the authors should present the recent advances for ABCG transporters in Introduction. Second, the authors should identify the ABC transporter genes in pigeon pea genome, and named as CcABCG1-CcABCG55. And then, genome-wide bioinformatics analysis of the CcABCGs, containing genome distribution, gene duplication, gene structure, evolutionary relationship, conserved motif, promoter analysis and putative expression patterns in different organs and under different abiotic stresses, can be further investigated to assess the importance of these genes in pigeon pea.

4. In method section, the authors claimed that they searched potential ABC proteins using the amino acid sequences of Arabidopsis ABCs as queries. In my opinion, it may omit some ABC proteins. The HMMER program should be better. PF00005: ABC transporter domain; PF01061: ABC-2 transporter domain; PF00664, ABC transporter transmembrane region domain.

5. The subsection “Expression analysis of the pigeon pea ABCG gene family in different organs and under different abiotic stresses” should be divided into two subsections.

6. Why did the authors only choose 10 ABCG genes for qRT-PCR analysis in different tissues and responses to various stresses? Please add more gene information about ABCG genes for qRT-PCR analysis. Segmental and tandem duplication genes may be better.

7. Some references listed in the manuscript are inappropriate. Many recent relevant reports about the ABC transporter family were even not mentioned.

8. The fact that statistical evaluation of the obtained results is missing.

9. The Discussion section does not present any real conclusions and comments. The authors should discuss more about the ABCG transporters in adaptation of pigeon pea to abiotic stresses, and the relationships of CcABCGs and other important ABCG transporters identified in other plants.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 2
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: The authors have added new explanations/experimental data in support of their claims and as a rebuttal to the reviewer’s comments. I am satisfied with most of the replies and additions the authors made. However, I still think the authors could concentrate the manuscript more on the effect of stress and gene regulation, rather than the chromosomal positions or intron-exon composition of the genes (authors have dedicated entire figures on them). The argument that having more exon make them prone to more regulation is theoretical and no evidence was provided in support of that statement. It is true that we are still learning new things about the mechanism and regulation of the splicing process and the statement could be theoretically sound but that still doesn’t carry any weight towards the conclusions of the paper.

This manuscript needs to be corrected thoroughly for linguistic issues (grammatical mistakes, sentence structures, typos, etc.) and rewritten for scientific clarity.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Thanks for the author's serious response. This revised manuscript has been substantially improved, and the questions that the authors have explained and the errors that the authors have corrected. However, there are still a number of grammatical mistakes, such as line 50, 240, 297, 341, 350, 391, 407, and so on. The authors should seek an English native speaker to correct the grammar mistakes for improving the readability. In addition, the authors should update the descriptions in panels (A-D, G and H) of figure 6. These errors should be addressed before publication.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 4
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: I am scientifically satisfied with the new changes. In terms of language, this manuscript is unfortunately still far away from perfection. There are typos and grammatical errors everywhere. Here are some (corrected) examples just to name a few--

Missing s for third-person singular subjects:
The NBD domain consists of three highly.., NBD domain on the cell membrane hydrolyzes ATP.., etc.

Oxford commas are missing:
In order to reveal the important role of ABCG transporters in resisting environmental stress in pigeon peas, identification, phylogenetic analysis, and expression analysis, etc.

Was/were issues:
gene localization was annotated..., Gene structure was analyzed based..., etc.

Missing auxiliary verbs:
Results have shown that the number of motifs in the WBC...

Is/are issues: The cis-acting elements are...

Typos: gametogenesis, ABCE and ABCF subfamily had no transport function..., etc.

Overall, The manuscript is ready to be published but only after thorough proofreading. I would recommend the authors to use some professional writing service before the final publication.


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

Input:
Title: THE FUNCTIONAL ANALYSIS OF ABCG TRANSPORTERS IN THE ADAPTATION OF PIGEON PEA (CAJANUS CAJAN) TO ABIOTIC STRESSES
Review round: 6
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: This manuscript has improved a lot. I, however, standby my previous recommendation that the authors may want to use writing services. There are many sentences that require restructuring. For example the first sentence of the Discussion section --

ABC transporters are distributed in animals and plants, but ABC transporters in plants are
characterized by a large number and complex function, such as 129,127 ABC transporters were
identified in Arabidopsis and rice.

I am not certain what message does this carry? These types of sentences are everywhere. I once again would request the authors to proofread the manuscript thoroughly.

Overall, this manuscript is ready to be published but only after thorough proofreading.


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: TRENDS IN THE INCIDENCE, PREVALENCE AND YEARS LIVED WITH DISABILITY OF FACIAL FRACTURE AT GLOBAL, REGIONAL AND NATIONAL LEVELS FROM 1990 TO 2017
Review round: 1
Reviewer: 1

Output:
Basic reporting: Authors have used clear and professional English throughout the manuscript. Authors have provided proper literature references. However, I think they could have elaborated more on the background/context. They could have emphasized why the study is important in more details. Article structure is professional. Figure legends could be made bit more legible. Tables are very professional. The manuscript is self-contained with relevant results to hypothesis.
Experimental design: The experimental design is standard as expected in the field. The research question is well defined. Author have collected data from legitimate sources and performed proper statistical analyses. Methods have been described with sufficient details.
Validity of the findings: The findings of the manuscript are valid and will definitely help policymakers around the world. I really liked the way authors discussed the results and put it in the context.
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: TRENDS IN THE INCIDENCE, PREVALENCE AND YEARS LIVED WITH DISABILITY OF FACIAL FRACTURE AT GLOBAL, REGIONAL AND NATIONAL LEVELS FROM 1990 TO 2017
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is written in professional, Scientific English. The introduction just sufficiently sets the theme of the study and is well-referenced.
However, all the figures and tables miss the description and headings which makes it very difficult to follow. Please label them.
Fig-1 The legend is not clear
Fig-2 What is the difference between graphs 2A, 2B and 2C?
Fig-3 What is the difference between graphs 3A, 3B and 3C?
Fig-4 Labels A,B, C missing.
Fig-5 Please explain how the correlation between EAPC and SDI is different among Fig. 5B, 5D and 5F.
What is the difference between Fig-1 and Fig-6? They look exactly the same.
Fig-7 misses description.
Fig-8 Which trend does each graph represent?
Fig-9 Fig 9B and 9C are the same.
Fig-10 misses description.
Experimental design: The research question is well-framed. Though the data source is mentioned in the abstract, the methods section is missing. Thank you for providing the raw and processed data. Could you please:
1. Describe the methodology used to analyze the data and draw statistical inference for YLDs, SDI, and APRs?
2. How was EAPC calculated?
3. Why did you choose data only till 2017? Although the latest 2019 report of Global Burden of Disease is available.
Validity of the findings: The study is exhaustive and provides all underlying data. The study is presented in a tabulated form region-wise, making it easier to follow.
Please state explicitly how this study would directly impact the medical resource allocation and policymakers as stated in the introduction (provide if you have any correlation data).
Additional comments: No comments


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

Input:
Title: TRENDS IN THE INCIDENCE, PREVALENCE AND YEARS LIVED WITH DISABILITY OF FACIAL FRACTURE AT GLOBAL, REGIONAL AND NATIONAL LEVELS FROM 1990 TO 2017
Review round: 1
Reviewer: 3

Output:
Basic reporting: The figures should be made in a more representable form.
Experimental design: The methodology section is missing throughout the manuscript.
Validity of the findings: No comments
Additional comments: The manuscript “Trends in the incidence, prevalence, and YLDs of facial fracture at global, regional, and national levels from 1990 to 2017” is a good compilation of data from 1990 to 2017 to understand the overall health burden of facial fractures. The study could be useful to the countries to make policies to reduce the occurrence of these events in the society. The study provides a thorough in-depth analysis of the available data. However, there are few concerns which needs to be addressed:

General Comments:
1.	The author should make a separate section for describing the method used. This section should be separate from method described in the abstract.
The authors should not use an acronym in the manuscript title. The authors have extensively used acronyms throughout the manuscript, most of the time it makes it difficult for the reader to follow. Overall, the authors need to use less acronyms.
2.	The text of the manuscript has some formatting errors.
Line 254 “this first study was the to systematically¨
Line 229 “leading to a high death rate in (Toivari et al. 2016)
Line 62 and 71 GBD acronym should be introduced in line 62 rather than 71

3.	In addition, the authors need to make figures in a more representable form for better understanding. The text of the labels needs to be enlarged For eg.; figure 2 need to be magnified.

Specific comments:

1.	The full form of ASIR has not been mentioned in the manuscript but used frequently by the authors in the complete manuscript.
2.	The labels of figure 4 are absent.
3.	The legends of figure 9 does not match with the figures. Authors should check these errors.


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: TRENDS IN THE INCIDENCE, PREVALENCE AND YEARS LIVED WITH DISABILITY OF FACIAL FRACTURE AT GLOBAL, REGIONAL AND NATIONAL LEVELS FROM 1990 TO 2017
Review round: 2
Reviewer: 1

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


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

Input:
Title: TRENDS IN THE INCIDENCE, PREVALENCE AND YEARS LIVED WITH DISABILITY OF FACIAL FRACTURE AT GLOBAL, REGIONAL AND NATIONAL LEVELS FROM 1990 TO 2017
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript is well written. The results and conclusions of the papers are interesting and convincing. I also find the analyses to be relevant and well documented. Statistical analysis is thorough and reliable.
Experimental design: The manuscript is well designed.
Validity of the findings: No comments.
Additional comments: Dear Authors,
Congratulation for this interesting study. The manuscript by Wu et al. reported the systematic assessment of global burden of facial fractures across all age and sex group. I have some suggestions for the authors.

1.	The authors may consider to add a table depicting sources of clinical records used calculating cause-nature proportions for facial fractures.
2.	What is the basis of selecting the 1990 as starting year?
3.	This study reported that- the ratio of facial fractures between males and females was 2:1. What could be the reason?


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

Input:
Title: GENOME-WIDE ANALYSIS OF GROWTH-REGULATING FACTORS (GRFS) IN TRITICUM AESTIVUM
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this reasonably well written manuscript, Huang et al. describe the identification and bioinformatics analysis of the GRF family in Triticum aestivum. In addition, they analyzed their expression patterns under drought and salt stress, which will facilitate further study on the relationship between GRF gene family in Triticum aestivum and its development regulation network and stress response. They have used clear and professional English throughout the paper.

Major issues:
1.Figure 3: In the legend to the Figure, the author declares that the neighbor-joining method is used, but in the manuscripts, he writes Maximum Likelihood method (Line 103). The two are inconsistent. The author needs to clearly use the method, and the phylogenetic relationships generated by different methods may differ significantly.

2.Figure 3: The author needs to be in the analysis of evolution bootstraps values marked in the Figure, high bootstraps values on the branch gene deserves more attention, at the same time can also be found TaGRF genetic relationship with the distance of GRF gene in different species.

3. Figure 3 and Figure 1: In the two Figures, the author claimed that both phylogenetic relationships were constructed by neighbor-joining method, but the results of the two Figures were inconsistent. Assuming that GRF genes of other species in Figure 3 were excluded, the evolutionary relationship of TaGRF in Figure 3 was inconsistent with that in Figure 1.

4.Line 286-289: The authors speculate that the conclusion that TaGRF family plays an important role in wheat shoot tip meristems and salt stress is not rigorous enough. In Figure 7, only a few TaGRF genes show significant changes in expression under the above conditions, which is not enough to indicate that the whole GRF gene family has the same role.

5.Figure 7 and Figure8: The author should link some important genes in the two Figures, such as TaGRF1-2D, TaGRF4-4A and TaGRF4-4D. The above-mentioned genes play an important role in the tissue expression pattern and salt stress in FIG. 7, but unfortunately the author did not correlate them with the transcriptome data in FIG. 8, and further explain the problem.
Experimental design: 1.The authors should use bioinformatics methods to identify several highly reliable TaGRF genes, otherwise the experiment will not be accurate and definitive conclusions.
Validity of the findings: No.
Additional comments: minor issues:
1.Line 97-99: It is suggested that the authors submit the protein sequences of candidate genes to the NCBI CDD (https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi) and SMART (http://smart.embl-heidelberg.de/smart/set_mode.cgi?NORMAL=1) databases for the presence of the conserved domain of GRF.

2. The author needs to explicitly refer to the source of the software used, such as Line 134, TBtools software.

3. Figure 1：It is recommended that the authors rename motif 1 and 2 to WRC and QLQ to be more intuitive.

4.Line 89: of and Pfam lack of space.

5. Some punctuation is not standard and English spelling problems.


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

Input:
Title: GENOME-WIDE ANALYSIS OF GROWTH-REGULATING FACTORS (GRFS) IN TRITICUM AESTIVUM
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this study, the wheat GRF gene family was identified and comprehensively analyzed including phylogenetic evolution, gene structure, and gene expression.
This has not been reported in wheat, which will provide a theoretical basis for the study of plant development and stress. The overall experimental methods and strategies are correct, and the experimental content is complete. However, the structure and logic of the entire manuscript need to be improved, and the expression should be more professional.

Introduction part:
Abbreviation should be interpreted where it first appears, (Gln, Leu, Gln) and (Trp, Arg, Cys) should be placed in line 41 instead of lines 42 and 44.

Lines 51-53 need inferences.

I think the functions and mechanisms of GRF participation should be listed in more detail and more organized. In lines 51-61, you explained GFPs' participation in the formation of plant tissues or organs, but there was no turning point or summary when you cited other functions of GFP in lines 62-66. In order to laying the groundwork for your research, you should describe its possible mechanisms in more detail rather than simply describe the functions in defense responses (lines 63-66). Also, pay attention to English expressions.

GRF TFs not only participate in plant growth and development, but also respond to certain abiotic stresses (Kim et al., 2012).
It is more appropriate to put this sentence in the previous paragraph than here.

It is better to put a separate paragraph on the brief introduction of content of this study (lines 76-82) and it should briefly introduce the significance and value of this research.

Material and Methods
The e-value of hmmer search should be given (line90).

In lines 97-99, how to eliminate sequences which did not belong to the GRFs? Exclude sequences that do not contain both WRC and QLQ, or sequences that contain one of the domains are also excluded?

In lines 112-117, the evaluation criteria for duplication events are strict and reasonable, but here you need to add what strategies and software were used to obtain tandem and segmental duplications.

Lines 141-145, The SRA ID of the RNA-seq data should be given, the version and references of the software used should be given.

Four Species ? The orthologous genes from T. urartu, T. dicoccoides, and Ae. tauschii were identified by reciprocal BLASTp analysis.

There are no problems with the method used in this study, but there are two major deficiencies in the presentation in this section. (a) The content arrangement is a bit messy and should correspond to the result part. For example, ‘Characterization Prediction of TaGRF Proteins’ was placed in the fourth paragraph, but in fact it was described before you described duplication events in the results section. Similar problems need to be modified. Individual paragraph content can be merged into one paragraph. (b) Language description should pay attention to avoid Chinglish. Sentence-by-sentence description of some content will be ambiguous, you can merge two sentences into one by using clauses, etc. E.g, lines 130-131. In addition, the subtitle expression has grammatical problems, especially the following three.
Sequence Alignment and Construction of Phylogenetic Tree
TaGRFs Expression Profiling by Mining Multiple Transcriptome Data
Wheat Seedlings Growth and Stress Treatments

Lines 184-186, what is the result? How many TaGRF you identified ? In addition, you used two strategies instead of only blastP search.

Line 186, TaGRF should be shown in Italics when it present gene.

Line 197, former grouping rules, what does it mean? References should be given.

Line 210-211, to supplement the method of multiple sequence alignment in material and methods part.

It’s reasonable to put phylogenetic analysis before structure analysis. Otherwise, how to explain the basis of the evolutionary tree in Figure 1? In fig 2, it is better to show phylogenetic tree in a suitable appearance for easier to find their corresponding names for some short branches.

It’s reasonable to put chromosome localization analysis after identification of TaGRFs, because you renamed them according to chromosome localization. It is also not very appropriate for two sentences to form a paragraph. Don’t show the duplication events in fig5.

Lines 250-262, this content should be put in discussion part.

Line 249-277, this content should be followed after identification and phylogenetic analysis (with Arabidopsis, rice, and maize). The subtitle is inappropriate and the results should be properly detailed.

Discussion
The discussion section should be carefully revised.
This part should be combined with the results of this study and existing research for discussion, rather than use a super paragraph to describe the experimental results in isolation, and another paragraph to introduce future research.

Figures and Table
The legends of fig 4, fig 5, fig 6 and fig 8 should be more detailed.

In this study, the related information of signal peptide should be removed because no result have been obtained.