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: EXPRESSION OF IER3 IN HEPATOCELLULAR CARCINOMA: CLINICOPATHOLOGY, PROGNOSIS, AND POTENTIAL REGULATORY PATHWAYS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Whether the statement of“IER3’s ？？？ potential molecular mechanism" in 306 is proper?
"expression levels in HCC tissues was ??? associated with poor OS." in line 367
"When RFX5 is??? silenced, it could significantly downregulate the protein levels of" in 393
”high IER3 expression was related ???  poor prognosis in HCC patients.“ in line 407
Please check the whole text.
Experimental design: "Figure 4 is Uni-/Multi-factor Cox analyses" in the rebuttal letter.
I did not find the cox information in Figure 4.

"the correlation between IER3 and RFX5 should be analyzed in cellular experiments."
The question is not answered directly.
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: FIRST OBSERVATIONS OF OVARY REGENERATION IN AN AMPHIPOD, AMPELISCA ESCHRICHTII KRØYER, 1842
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment - I find the manuscript meets these standards.
Experimental design: Some methods and logic underlying experimental design should be clarified, as noted in the full report.
Validity of the findings: Findings are well supported by the data provided, but their interpretation should be softened in some cases, as described in the full report.
Additional comments: Summary
This manuscript presents work inspired by the observation, made in previous studied by these authors, that female Ampelisca eschrichtii amphipods, appear able to restore ovaries to full function following starvation-induced ovarian atrophy. Such females, which would presumably normally lose reproductive ability at two years of age, had been observed to occasionally give rise to offspring at three years old. The authors ask what the source of “new” germ cells and somatic cells of the ovary might be in these females, and use classical histological techniques to address this question. They interpret their observations as suggesting that in the “regenerated” ovary, germ cells arise from previously dormant early stage oogonia, and that somatic cells arise from mesodermal cells that migrate to the ovary from their origins in the intestine.

The manuscript is clearly and concisely written, the experimental design is quite clear and sound overall, and the data of good quality and well presented. The problem addressed is extremely interesting and important for reproductive and regenerative biology in general. My only major concern is that some of the conclusions presented, namely those concerning the migration of cells, are overinterpretations of the data available, which are static snapshots of fixed tissue. It is understandable that the data are limited in this way, given the apparent rarity of this type of regeneration and the limited samples therefore available. This issue could easily be addressed by simply acknowledging in the manuscript the limitations of the data type available, and discussing alternative explanations for the observations shown.

Other minor concerns are also all easily addressable by simply adding clarifications to the manuscript, including (a) the relationship between animal length and age; (b) the evidence that animals collected from the field for observation are indeed “starving”; (c) the cytological criteria used to distinguish different cell types from one another.

Below I provide numbered explanations of these most important points; other minor suggestions for the MS are available in the marked PDF that accompanies this review.

Major Concerns
1.	It is not possible to unambiguously conclude that any cells are migrating, from data that are gathered from fixed samples. Please address this in the manuscript, and soften or modify conclusions presented accordingly.
2.	In a number of places in the manuscript, single examples of cellular behavior from the literature are stated as if they were universal features across animals. Examples include text in the areas of lines 71-77 and lines 218-227. Please rephrase these sections of the manuscript to make it clear in which animal(s) or cell type(s) there is evidence for these processes, in a way that accurately reflects the references cited as evidence for these statements.

Minor Concerns
1.	Body length is taken as a proxy for age, but there may be limitations to this approach especially if animals are indeed starved for any period of their life. The authors should make this caveat explicit.
2.	Ovary regeneration is taken to be a possible response to starvation conditions, but the evidence that the animals observed by the authors were actually starving is unclear. The authors should make this caveat explicit.
3.	The criteria used to designate cells as “eosinophilic” is not clear from the Figures. For example, in Figure 1, the legend states that mesodermal cells are “intensely stained” with eosin, “in contrast” to follicular cells,” but in the micrographs shown, the cells labelled as follicular cells have darker staining than those labelled as mesodermal cells.


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: FIRST OBSERVATIONS OF OVARY REGENERATION IN AN AMPHIPOD, AMPELISCA ESCHRICHTII KRØYER, 1842
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article meets standards of professional English. Just minor spelling and grammatical errors were observed. Literature references were sufficient.
Experimental design: The finding about this interesting life history story of extended reproductive success under pressure from likely limited resources the previous breeding season is interesting. A great example of how an animal can cope with environments with limited resources. I include in the comments below requests for further description of the histological methods and some brightening up of the images.
Validity of the findings: The study is observational in nature, which was done so sufficiently.
Additional comments: Below are several specific comments on errors or requests for further explanation.

Line 58 and 63: Epithelium misspelling

Line 59-61: Rework sentence structure

Line 106: Misspelling of formation

Line 123: Rework sentence structure

Methods: More information on the histological preparation is needed. Are these cryosections or paraffin sections. What was the tissue section thickness. Were all the stains hematoxylin and eosin stained and how was this performed?

Figures: The histological images could be brightened up so the background is white. This will provide more contrast for the reader, which is acceptable because it shouldn't change change any of the interpretations or conclusions made from the images.

Line 141-143: Rework sentence structure

Figure 2: How many cells were visualized?


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 THE DIAGNOSTIC AND PROGNOSTIC VALUES OF SERUM HSP90Α IN SEPSIS PATIENTS: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript of a good piece of work and adds significant information to the literature. The article structure is good and follows the journal`s pattern and guidelines. The hypothesis or study questions are well elaborated and tested. Thanks to the authors for sharing the raw data.
Experimental design: Thank you for providing the information on the ethical context of the manuscript. Since the authors adopted a case-control approach in this study, I did not find any information on sample size estimation? The representation of the sample for other settings is quite valuable, authors should discuss this in limitation if sample size estimation was not done.
It will be also valuable for readers to get details of the hospital in which this study is conducted i.e., type of hospital, the number of patients received or covered, no. of beds and types of services etc.
Please also include the normal values used in the current hospital for HSP90α or other biomarkers in the methodology section.
Validity of the findings: The authors have vigorously tested the relationship of HSP90α with sepsis and mortality. In the abstract section, the AUC of HSP90α was 0.8 while it has a different value in Figure 2.
Table 1 provides information on three arms of the study. Since the study design was case-control and the authors have matched all three arms at the time of patient selection, I did not see a statistical comparison of demographics between these three arms.
I have also observed that there are some missing data in the analysis, but the information of treating the missing values is not provided in the methodology section?
Table 1: 56/94???? what does it indicate, or the data was only available for 94 patients out of 150 septic patients? or this 94 are the patients who survived? the similar confusion will be created among readers so it is advised to please consider the elaboration of results presented in the table carefully. Technically, I will estimate the percentage of mortality i.e. 56/150 septic patients. How the data is presented in the table must be described in the footnotes of the table.
I am impressed by the logistic models in this study. However, I would suggest having values for univariate too for each included variable. If authors agree on that, they can provide univariate odds ratio and p values for each of the included variables in text and then can provide detailed information on the logistic regression.
Tabel 3: the information on Model 5 is not provided in the footnotes. I have also one concern here, the authors have aggressively analyzed the relationship between various markers with sepsis but the association of comorbidities is also questionable in this regard. Can authors discuss this in the discussion section to rule out any confounding impact of comorbid conditions?
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: EVALUATION OF THE DIAGNOSTIC AND PROGNOSTIC VALUES OF SERUM HSP90Α IN SEPSIS PATIENTS: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript entitled “Evaluation of the diagnostic and prognostic values of serum HSP90α in sepsis patients: a retrospective study” by Fuxing Li et al. investigated whether heat shock protein 90 α (HSP90α) can be used as diagnosis and prognosis markers in patients with sepsis. They have found that 1) serum HSP90α in sepsis patients were higher than those in ICU and healthy controls; 2) HSP90α was an independent risk factors for sepsis mortality; 3) HSP90α is a prognostic marker of sepsis.
Experimental design: In Figure 8, HSP90α is highly correlated to cytokines and chemokines (IL-1β, IL-18, and MIP-3α). If these factors along with HSP90α are assembled into a biomarker signature/model, will it demonstrate a better performance (increased AUC and OR) to diagnose and predict prognosis in septic patients? Authors can also test the performance of HSP90α combined with Crea, BUN, SOFA score and PCT.
Validity of the findings: 1.	In Figure 4, five variables were shown as significant risk factors for the survival of sepsis patients, including PCT, SOFA, creatinine (Crea), blood urea nitrogen (BUN) and HSP90α. Although ORs were statistically significant, how is it clinically important with an OR that is almost equals to 1? After adjusting multiple variables, OR was almost 1, as shown in Table 3.
2.	To support the potential clinical application of HSP90α, the cut-off value (120 ng/mL) should be validated in independent patient cohorts
3.	Authors indicated that HSP90α affected the prognosis of sepsis patients via provoking the secretion of pro-inflammatory cytokines and chemokines. However, the correlation between HSP90α level and inflammatory cytokines cannot support the conclusion that HSP90α induces the expression of cytokines thus leading to sepsis.
Additional comments: In text line 28, “blockage of HSP90α could attenuated the inflammatory damage”. It should be “attenuate”.


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 THE DIAGNOSTIC AND PROGNOSTIC VALUES OF SERUM HSP90Α IN SEPSIS PATIENTS: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: All the comments are addressed by the authors.
Experimental design: All the comments are addressed by the authors.
Validity of the findings: All the comments are addressed by the authors.
Additional comments: All the comments are addressed by the authors.


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 THE DIAGNOSTIC AND PROGNOSTIC VALUES OF SERUM HSP90Α IN SEPSIS PATIENTS: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Authors have successfully responded to all my comments.
Experimental design: None.
Validity of the findings: None.
Additional comments: None.


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

Input:
Title: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. The English language was so bad, and it should be improved to avoid Chinglish-expression forms and grammatical errors in the whole manuscript. I suggest that the author should have a colleague, who is proficient in English and familiar with the and familiar with the subject matter, to review your manuscript, or contact a professional editing service to revise it.

2. I have several criticisms to this manuscript for the messy format of the papers, such as the “left-aligned text”, the “random space” in the line 55.

3. There exist some mistakes in manuscript, such as the figure legends of Fig. 2, which does not match up with the Fig.2A.

4. The authors were not able to describe these results successfully, besides, they also could not explain about the cross-talk between ARFs and leaf development
Experimental design: No
Validity of the findings: No
Additional comments: No


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: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper demonstrated the cloning of ARF1 from Cymbidium goeringii and ectopic overexpression of CgARF1 in Arabidopsis plants. I have many concerns on Figures and methods. Especially, essential data, and information are missing in legends and main text.

1) The description of Figure legend should be improved in all figures. The essential informations were not mentioned in Figure legends. For example, In, Fig. 5A and 5B, which group represents WT or transgenic plants?

2) Figure legends are missing in all supplemental Figures.
Line 207-209 Authors compared CgARF1 and 19ARFs (Fig. 1 and S2). However,the plant species and gene/protein ID were missing in Figure 1 legend.

3) Statistical analysis procedures are not shown in Figure. Which tests is applied to data, Post-hoc test, Tukey-Kramer test, Bonferroni test, Dunnet test etc, please also specify the sample number in each figure panel. Please specify the number of biological replicates and technical replicates in the methods section or legends..

4) Bar graphs of Fig. 2A and 2B are incorrectly displayed. Fig. 2A and 2B should be exchanged each other.

5) In the methods section， Line 162
The ORF of the CgARF1 gene (without a stop codon) was inserted into the pCambia1300 vector (Table 1), which contains the green fluorescent protein (GFP) reporter gene driven by the CaMV 35S promoter. Please describe the correct methods of plasmid construction. Thus, the control of GFP vector was not indicated. In the text (line 231), CgARF1::GFP fusion protein was described. The position of GFP fused with CgARF1 protein (N / C terminus) was not specified according to the method section (pCambia1300 vector).

6) pCambia1300 vector does not contain GFP under 35S promoter.
https://www.snapgene.com/resources/plasmid-files/?set=plant_vectors&plasmid=pCAMBIA1300

7) Line 70 However, all ARF proteins do contain all three complete domains, as is the case for AtARF3/7/13/23 in Arabidopsis (Guilfoyle et al., 2001; Remington et al., 2004) and CiARF3/14/17 in sweet orange (Li et al., 2015).

“all ARF proteins do contain all three complete domains” is correct? Probably, all ARF proteins do NOT contain?

8) “as is the case for AtARF3/7/13/23 in Arabidopsis” is not correct. AtARF7 contains three complete domains. Probably, ARF7 >ARF17.

9) Line 77, AtARF7 and AtARF9 are also induced in senescent leaves (Lin and Wu, 2004). This sentence would not be appropriate, because the cited paper did not describe the ARF expression in the text although raw data might contain the expression of ARF7 and ARF9.

10) Line173, Author constructed pBI121-35S::CgARF1. pBI121 is 35S::ORF-GUS vector. Is the CgARF1 protein expressed as GUS-fusion?

https://www.snapgene.com/resources/plasmid-files/?set=plant_vectors&plasmid=pBI121

11) Line 191  MS medium with 0.2 mg·L-1 or MS medium with 0.5 mg·L-1. > MS medium with 0.2 mg·L-1 IAA or MS medium with 0.5 mg·L-1 IAA

12) Line 238-241
The CgARF1 overexpression in transgenic lines should be confirmed by qRT-PCR. The genomic PCR analysis did not show any evidence of CgARF1 expression in transgenic lines

13) Line 260 Changes in endogenous phytohormone contents under IAA treatment

13-1) Please describe full details of phytohormone measurement by ELISA. Please indicate the commercial vendor of mAb for each hormone or ELISA kit if used (code number and vendor) .

13-2) Line 264, After 12 h of treatment, exogenous IAA treatment caused a substantial increase in endogenous IAA levels. The IAA level in CgARF1-overexpressing plants began to decline after 12 h, but the level in WT plants remained high.

> I could not understand why IAA level is measured after exogenous IAA treatment. exogenous IAA and endogenous IAA could not be distinguished by ELISA methods. If synthetic auxins NAA or 2,4-D are used, IAA might be measured by ELISA. However, exogenous IAA was used in this experiment. The authors would like to examine the rate of IAA inactivation between WT and CgARF1-overexpressing lines? Discussion (lines 319-323) should be improved.

13-3) Line 261 and 266, The contents of endogenous hormones, including IAA, ABA, GA3, and BR,
Fig. 6B-D shows that the contents of other endogenous hormones in transgenic plants were almost all lower than those of WT plants, especially for GA3.

>Plant does not produce GA3 as endogenous active GA, GA1, and GA4 are endogenous gibberellins, GA3 is produced by the fungus.

13-4) Line 264 exogenous IAA treatment caused a substantial increase in endogenous IAA levels.
>Please indicate the method for exogenous IAA treatment and the concentration of IAA.
Experimental design: Experiments seem to be well designed and performed, but many errors are documented in the method section. The expression of CgARF1 was not confirmed by qRT-PCR. The insertion of CgARF1 gene into the Arabidopsis genome was confirmed by genomic PCR. This qRT-PCR data is crucial for the confirmation of CgARF1 overexpression.
Validity of the findings: I could not confirm all data in this study as the legends of supplemental Figures are missing.
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: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 1
Reviewer: 3

Output:
Basic reporting: This article studied ARF1 from Cymbidium goeringii. Researchers analyzed basic information of CgARF1 using straightforward bioinformatic tools and identified CgARF1 expression pattern in WT and IAA treatment. More important, through overexpression of CgARF1 in Arabidopsis, authors investigated the potential function of CgARF1 during leaf development under IAA treatment compared with WT plants. Overall, conclusions of this article are appropriately supported by well-designed experiments and comprehensive analysis.
Experimental design: Figure 2 is supposed to show the expression level of CgARF1 in different tissues during the time course and under IAA treatment as well. However, the data shown in Figure 2 could not reflect or support the explanation/ conclusion in Line 215-226. Please provide elaborate description of this Figure 2 regarding but not limit to the following concerns,
a.	which organ (R, P, L, F) is the expression level shown in Figure2A from?
b.	As indicated in Line 224, where is the data in Figure 2B showing 2h, 12h expression?
c.	It would be more supportive to include qRT-PCR raw data in supplementary set.
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised manuscript has been much improved and answered all my concerns. I have no more comments.
Experimental design: The revised manuscript has been much improved and answered all my concerns. I have no more comments.
Validity of the findings: The revised manuscript has been much improved and answered all my concerns. I have no more comments.
Additional comments: The revised manuscript has been much improved and answered all my concerns. I have no more 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: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 2
Reviewer: 2

Output:
Basic reporting: The revised manuscript is highly improved and satisfied all my concerns. I think that this round manuscript is suitable for publication in the PeerJ article.
Experimental design: The experimental design is appropriate to support the conclusion.
Validity of the findings: All provided data support the conclusion.
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: FUNCTIONAL ANALYSIS OF ARF1 FROM CYMBIDIUM GOERINGII IN IAA RESPONSE DURING LEAF DEVELOPMENT
Review round: 2
Reviewer: 3

Output:
Basic reporting: In the revised manuscript, authors resolved all concerns from my previous review. This article studied the properties and function of CgARF1 from C. goeringii. This study provides useful information as a preliminary investigation of ARF1 roles in leaf development. The underlying mechanisms still needs to be explored.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: EFFECT OF PRECIPITATION CHANGE ON THE PHOTOSYNTHETIC PERFORMANCE OF PHRAGMITES AUSTRALIS UNDER ELEVATED TEMPERATURE CONDITIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The research paper entitled “ Effect of precipitation change on the photosynthetic performance of Phragmites australis under elevated temperature conditions” has quantified an effect of precipitations on photosynthesis under elevated temperature.
The abstract of the paper is factual concrete, realistic, understandable, illuminates the main findings of the paper, and can serve as a stand-alone document that succinctly describes both procedures and conclusions.
The authors have put in effort to compile literature linking and highlighting the relation of photosynthesis under changing environmental and plant water regime. This part looks informative authors tried to write a good compilation of citations and review of the work, however, the majority of references are quite old and not very focused on mechanisms of photosynthesis. The philosophy of this work is good, applicable mostly in applied plant and agronomy sciences. The understanding of mechanisms is limited.
The paper might be a hot topic for the next experimental research in model crop plants, therefore, contributing to its merit for acceptance in PeerJ.
Experimental design: The experimental methodology was appropriate, enough strong and scientific, and the analyses were done correctly. The authors used very progressive techniques and protocols and described used protocols perfectly.
Validity of the findings: The novelty of this manuscript is not very high, however, the paper brings new aspects and news. I would like to encourage the authors to discuss more ecophysiological aspects mechanisms, including impact on plant photosynthesis and primary processes of biomass production. The authors should know about classical papers (Cornic G., Lawlor D.,  Chaves M.,  Brestic M.,  Loretto G., Zivcak M. etc.) about the flexibility of photosynthesis apparatus under drought stress and the stomatic and non-stomatic inhibition of photosynthesis, which is a key limitation until 25-30% of leaf relative water content.
The paper adheres to appropriate reporting guidelines and community standards for data availability and is presented in an intelligible fashion and is written in standard English.
Additional comments: Several sections in the manuscript lack experimental evidence as well important references of authoritative articles. Authors were unable to generate their strong own synthesis of information in a scientifically correct manner.  The authors could explain more about the mechanisms of plant sensitivity to water deficit. Authors could involve also new aspects of plant responses to water deficit and also modern phenotyping approaches used in precise agriculture, including new references.
In the current version, the review could be improved toward being more balanced, structured and critical. The authors have to discuss more in functioning photosynthetic apparatus in a changing environment.
The biggest gap in this paper is a lower understanding of the research of photosynthesis under stress.
The authors should improve the interpretations and discuss their results more properly.
Paper bring many new aspects and the novelty of the paper is OK, but I would like to invite authors to discuss more eco-physiological aspects and molecular mechanisms using new references:
•	Drought and Heat Stress in Cool-Season Food Legumes in Sub-Tropical Regions: Consequences, Adaptation, and Mitigation Strategies. Plants 2021, 10, 1038. https://doi.org/10.3390/plants10061038
•	Kernel Water Relations and Kernel Filling Traits in Maize (Zea mays L.) Are Influenced by Water-Deficit Condition in a Tropical Environment. Front. Plant Sci. 12:717178. doi: https://dx.doi.org/10.3389/fpls.2021.717178
•	Consequences and Mitigation Strategies of Abiotic Stresses in Wheat (Triticum aestivum L.) under the Changing Climate. Agronomy 2021, 11, 241. https://doi.org/10.3390/agronomy11020241
•	Photosynthesis research under climate change. Photosynth Res (2021). https://doi.org/10.1007/s11120-021-00861-z


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: EFFECT OF PRECIPITATION CHANGE ON THE PHOTOSYNTHETIC PERFORMANCE OF PHRAGMITES AUSTRALIS UNDER ELEVATED TEMPERATURE CONDITIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Please see the comments below.
Experimental design: Please see the comments below.
Validity of the findings: Please see the comments below.
Additional comments: Manuscript 67054v1

The manuscript reported the results concerning the changes in the photosynthetic traits in plants of Phragmites australis to water status change and high temperature. The main results are 1) increased precipitation can promote the photosynthetic capacity, whereas water deficit largely decreases it. 2) elevated temperature weakens positive effects of increased precipitation, and enhances the adverse effects of drought. 3) non-stomatal limitation can mainly contribute to carbon assimilation rate. 4) the xanthophyll cycle-mediated thermal dissipation may play a major role in PSII photoprotection, i.e., an adaptive traits. This study seems to be designed well, and the methods and data are appropriate. This main finding can be useful for the better understanding the plant stress physiology under the climatic change. Thus, it can be published. However, several issues should be addressed or revised as follows:

Abstract
‘To better understand how precipitation change and climate warming impact the photosynthetic performance of Phragmites australis (P. australis)’ is not the method topic, but object topic. It can be moved in the object part above. Or it could be changed accordingly.
“The combination of high temperature and precipitation change had more detrimental effect on P. australis than a single factor” seems to not be clear. What is precipitation change? it may be a deceased precipitation (drought)? Please change it.
In the section, the conclusion and/or application expressions seems to be lacked. Please add it.
Introduction
Line 2: “Global warming caused by high levels of greenhouse gas emission” can be change to “Global warming mainly caused by high levels of greenhouse gas emission”
Line 5: “IPCC, 2014” can be updated to the one “IPCC, 2021/2021?”. Please search, check and confirm it.
Line 21: “It’s widely accepted that”? “It’s” should not be used in its informal case, and may be changed to ‘it is’. Please check this case throughout the ms test.
Line 63-64: “Their spatial distribution is mainly limited by heat and water” seems to be not logical, and can be changed “Their spatial distribution is mainly limited by heat shortage and water deficit”?
Materials & Methods
Lines 78-79: How many year did the data obtain the annual average temperature and precipitation? 30 years? Please add this information.
Lines 87-96: some information could be added in details: such as light condition, the environmental chamber size.
Results
This section seems to be overstated a little bit, and it can be shortened to be concise. For instance, for photosynthetic parameters, lines 172-196, this part seems to be tedious, it can be shortened more to be readable more.

Conclusions
This part have main results, conclusion, and application expressions. However, the latter seems to be lacked. Please add this information.


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: EFFECT OF PRECIPITATION CHANGE ON THE PHOTOSYNTHETIC PERFORMANCE OF PHRAGMITES AUSTRALIS UNDER ELEVATED TEMPERATURE CONDITIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Yes.
Experimental design: Yes.
Validity of the findings: Yes.
Additional comments: The newly version has been prepared well, and the responses to the comments is ample. Thus, I can recommend it for publication after carefully checking it throughout.


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: FEATURE SCREENING FOR SURVIVAL TRAIT WITH APPLICATION TO TCGA HIGH-DIMENSIONAL GENOMIC DATA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Please see my additional comments.
Experimental design: Please see my additional comments.
Validity of the findings: Please see my additional comments.
Additional comments: The manuscript entitled “Feature Screening for Survival Trait with Application to TCGA High-dimensional Genomic Data” by Jie-Huei Wang et al. systematically reviewed multiple screening approaches and developed an optimal model for patient using TCGA genomic datasets with survival information. The study is of clinical significance with some minor issues.
1.	Does each biomarker gene have equal or different weight?
2.	How is the performance of biomarker genes identified in the manuscript to predict cancer patients’ prognosis compared to published gene signatures that predict survival?
3.	Authors could validate their biomarker genes in other independent human cohorts, like METABRIC.


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: FEATURE SCREENING FOR SURVIVAL TRAIT WITH APPLICATION TO TCGA HIGH-DIMENSIONAL GENOMIC DATA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors consider the problem of feature selection in the context of survival analysis using regression models.  The authors evaluate network-adjusted feature selection methods against several existing univariate feature selection methods on both simulated and real data.

Overall, I found the paper difficult to follow; reading the paper was much like being a detective trying to piece the evidence together, ensuring that I'm interpreting things as intended, and reverse engineering the story.  The paper would benefit from language edits as well as additional explanations.

The paper would benefit from substantial re-organization.  As it stands now, the paper is difficult to follow.  I would propose moving lines 147-185 to the Methods section.  It was not clear to me that the ESCA and PAAD results sections were subsets of the Real Data Application Section.  You may consider adjusting the font sizes or indicate this in one way or another.  Further, you should provide some introductory text in the section summarizing what data you will be analyzing before discussing the evaluation metrics.

Figures: I am not able to read the figure axis labels due to the low resolution and font sizes.  Please increase the resolution and font sizes.  Please spell out abbreviations such as MMS , provide a short explanation (e.g., 1 sentence), and indicate how to interpret the metric (e.g., small is better).  Please label the subfigures (e.g., a, b, c, etc.)  In the set similarity heat maps, the color bars indicate a range of possible values from -1 to 1, but Jaccard similarity only returns values in the range of 0 to 1.  Please update the range of the color bars.

The authors should cite sources for metrics such as the c-index.
Experimental design: Overall, I think this work has the potential to be very informative.  The overall setup of the experiments makes sense.  Overall, the authors’ main challenge is an over-emphasis on comparative (relative) metrics without a clear evaluation against ground truths, especially of the predicted survival times.  When applied to the simulated data, the NPN-MB method clearly recovers a larger proportion of the true predictors, results in models with fewer variables, and selects a different subset of features than the other methods.  The authors should indicate the impact of the feature selection on the survival time predictions errors.  For example, the authors should create additional plots like the first row of Figure 1 giving the c-index as a function of the number of selected features for each method.  The authors should also include the ground truth set of predictors in the set similarity analysis of features selected by each method.

That said, it was not clear how this paper is different from the authors' previous paper (Wang & Chen 2021).  If I am understanding correctly, the method being evaluated was actually introduced and already published in (Wang & Chen 2021) and evaluated on a subset of the TCGA data.  The current manuscript appears to build on the initial work by evaluating the method on additional data from the TCGA data set.  More evaluation is good, but the authors should clarify the relationship between the two manuscripts and the intent of the work in the current manuscript.
Validity of the findings: In the applications to real data, the authors evaluate the models based on five metrics, three of which are different ways of testing that the predicted survival times of the groups are different statistically.  Those three analyses are somewhat meaningless, however, and thus, distracting.  The important metric is the c-index, which compares the ranking of the subjects by their predicted survival times with rankings from their true survival times.  For the ESCA TGSA data set, the c-index indicates that the ordinary MCP model (0.8466) using all of the variables is more accurate than any model constructed using the feature selection techniques.  Secondly, The c-index score of the NPN-MB method (0.7380) is very similar to that of the CINDEX univariate feature selection method (0.7324).  It is not clear that feature selection NPN-MB produces a model with more accurate predictions than other feature selection methods.  That said, however, the authors’ main focus does not appear to be the creation of more accurate models but on the selection of informative features.  The authors analyze genes identified by their feature selection method and cite independent studies that corroborate some of the results.  This analysis indicates that NBN-MB is finding biologically-plausible (meaning, informative) associations.  Without a similar analysis of features selected by the other methods, however, we cannot concluder whether NBN-MB is better or worse at finding informative features than any other method.

As it currently stands, I do not believe that the conclusions are well supported.  The authors will need to perform a major iteration on their analyses, especially with respect to the choice of interpretation of the chosen metrics.  That said, I do think the method is interesting and a valuable contribution, so the evaluation on additional data sets as reported in this paper has potential.  With appropriate revisions, I would not hesitate to recommend acceptance.
Additional comments: Thank you for the opportunity to read about your work.  It is very interesting!  I look forward to reading a revised and improved version of the manuscript.


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

Input:
Title: FEATURE SCREENING FOR SURVIVAL TRAIT WITH APPLICATION TO TCGA HIGH-DIMENSIONAL GENOMIC DATA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Authors have successfully responded to all my comments.
Experimental design: None.
Validity of the findings: None.
Additional comments: None.


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

Input:
Title: FEATURE SCREENING FOR SURVIVAL TRAIT WITH APPLICATION TO TCGA HIGH-DIMENSIONAL GENOMIC DATA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The overall structure and organization of the paper has been improved considerably.  The paper is much easier to read and clearer.  The authors should be commended for their work.

At this point, I am mostly finding small typos and grammatical errors.  I would encourage the authors to have someone proofread the manuscript.

The paper does include a large number of figures.  Some of these figures can be combined.  For example, Figures 1-5 should be combined into a single multi-panel figure and Figures 6-10 should be combined into a second multi-panel figure.

Figures 1 and 2 and then Figures 6 and 7 are somewhat redundant.  The overlap coefficient is a a normalized version of the metric from Figures 1 and 2.  That said, I think that the overlap coefficient is more informative.  You could remove Figures 1 and 6 and specify the total number of true predictors in the figure legends so that the reader has infer the number of selected features from the given overlap coefficients.
Experimental design: The methods and results sections have been vastly improved.  It is much easier to read and provides more details necessary to understand and replicate the experiments.  The authors should be commended for their improvements there.

I am still struggling with the presentation of the results for the real data.  I believe that the evaluations of the simulated and real data should be as consistent as possible.

My understanding is that you are trying to find parsimonious models.  Meaning, if you models trained on two sets of features that have the same prediction accuracies, you want to select the model that uses feature features because the smaller number of selected features is easier to interpret / evaluate in follow up studies about biological function.

The choice of metrics and how they are currently presented for the real data does not make it easy to compare the feature selection methods.

My understanding is that the methods are run with a given parameter value to select features. A Cox regression model is trained on the chosen features and the accuracies of the models' predictions are evaluated.  Thus, there should be the following possible questions and evaluations:

- How accurate are the survival rate predictions compared with the ground truth values in absolutely terms?  E.g., if 80% of the patients survive into month 10, then the model predictions should match that.

	- I do not know enough about the data sets being used to know if this can be evaluated.

- How accurate are the survival rate predictions compared with the ground truth values in relative terms?  E.g., if patient A survives longer than patient B, then patient A should be ranked above patient B according to the predicted survival time.

     - It is my understanding that the c-index metric is the only metric that answers this question.  The other metrics (e.g., statistical tests) used only compare how well the models separate the two groups (good, poor) – this is not really useful and can be answered from the c-index metrics if you report the c-index for each class (good, poor) individually and together (so 3 c-index values per feature selection method per data set).  The other metrics reported just serve to confuse the reader.

     - Likewise, Figures 11 and 12 is not a very useful figure since the ground truth survival curves are not plotted alongside the predicted curves.

- How sensitive and precise is the feature selection?

       - E.g., how many of the true predictors are chosen?  To adequately evaluate this, you would need to know the true predictors.  This is probably not available but if you find papers that support the roles of specific genes, you can at least evaluate how many of those genes are found among the selected features.

       - E.g., I can do no feature selection and get perfect sensitivity as above.  Ideally, a feature selection method finds all ground truth predictors and none of the other predictors.  Again, since you may not have a list of all biologically-relevant predictors, you may only be able to report the number of features selected by each method.

      - Both of these metrics (precision, number of selected features) should be reported in tables 2 and 4 alongside the c-index values.
Validity of the findings: The discussion and conclusion section text has also been vastly improved.  The authors should be commended for their improvements there.

I believe that the findings (as currently presented) do support the research question.  Without the changes I have requested, however, the results for the real data are somewhat confusing and hard to navigate, making it difficult to be sure.
Additional comments: The authors have greatly improved the overall readability of the paper.  I enjoyed reading this revised version quite a bit.  I think there are still some important changes to make but these are not insurmountable.


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: FEATURE SCREENING FOR SURVIVAL TRAIT WITH APPLICATION TO TCGA HIGH-DIMENSIONAL GENOMIC DATA
Review round: 3
Reviewer: 1

Output:
Basic reporting: - The text (e.g., axis labels) in the figures need to be increased to be legible.
- The DPI of the figure images needs to be increased to make them legible
Experimental design: - no comments
Validity of the findings: - no comments
Additional comments: I want to thank the authors for addressing all of the comments in my previous review.  The figures are much easier to interpret and read.  The results are much clearer.


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: THREE NEW SPECIES OF MICROLAIMUS (NEMATODA: MICROLAIMIDAE) FROM THE SOUTH ATLANTIC
Review round: 1
Reviewer: 1

Output:
Basic reporting: English is generally correct and not ambiguous.
New features and characteristics are well and detailed described, but it is not always possible to check them because raw data are not submitted and some photos are blurry. To get high quality photos of meiofaunal animals is not always easy but it is an important goal in research diversity studies.
Drawings are clear and well done.
Symbol legends can be added to all the photos/drawings in order to help the reader finding the described features.
The submission is self-contained and the results truly clear.
Adding a Conclusions section can help the reader to understand why your research should matter.

20 An amendment to
21 the diagnosis of the genus is proposed.
Can you explain why you propose an amendment of the genus?

35 it was the most abundant genus, in terms of the number of individuals, of the family
How many individuals were found?

58 Although different in surface area, the two samplers have a similar design that aims
59 at maintaining the integrity of the vertical sediment structure
The meaning is unclear, can you explain this part?

A general background/context is provided.

26 (1922). This group is predominantly marine, although its type species M. globiceps De Man,
27 1880 is one of the few freshwater species.
M. globiceps is a terrestrial, marine and freshwater species, as many others (Nemys - https://nemys.ugent.be/aphia.php?p=taxdetails&id=121080 )

63 Each sediment sample was transferred to a plastic flask and fixed in 10% formalin
64 buffered with borax.
Adding a reference for this part can be useful in understanding the methodology.

349 sperm 350 present in uterus. How can you determine the presence of the sperm?
Experimental design: This research is in line within Aims and Scope of the journal. Questions are well defined and significant and results contribute in filling a knowledge gap in nematodes taxonomy.
The research is clear but, in some parts, experimental methods can be described more in detail providing references that can help the reader and potential replicators.

58 Although different in surface area, the two samplers have a similar design that aims
59 at maintaining the integrity of the vertical sediment structure…
and
62 three replicated sediment quadrats (10 x 10 cm) for meiofauna (0–10 cm depth) were
63 sampled.
Samplers or quadrats or both methods were used? How? To add references and explain how the quadrats were used can help in understanding the methodology.

65 Meiofauna samples were sieved through a 500-μm mesh…
A 500-micrometers mesh sieve is inadequate considering that nematodes are commonly bigger. A 1 mm or 2 mm mesh sieve is usually used instead as in ‘Influence of mesh size and core penetration on estimates of deep-sea nematode abundance, biomass, and diversity’ D.Leduc, P.K.Probert, S.D.Nodder
Can you explain why you choose to use this kind of sieve?
Validity of the findings: There are some features not identifiable in the photos, as for the cephalic setae structure.
In all images and photos adding abbreviations can be useful to speed up reading.
This is a suggestion: I think it is better to include all the features mentioned in the paper in the relative table for the same reason as before.

TAB.1
t or T and v or V (check in all tabs)

TAB.2
Use sign = after NA and NO instead of commas as in TAB.4 and TAB.6
Check significant figures (in all tabs)
Add cs/hd

TAB.3
Add space between ‘and’ and ‘morphologically’

TAB. 4
Add mbd/hd, hd/cs, t/abd because are mentioned in the description (same considerations for female allotype)
cs(=7,2) is not 50% of hd(=12,6)

TAB.5
For M. monstrosus write the length range 1104−1537 μm instead of 1240 as for the other examples

TAB.6
Add mbd/hd, cbd/cs (male), cs/hd (female)

TAB.7
Add amph. pos/hd = 0.4–0.7 in M. vitorius vs 1.5–1.7x in M. cyatholaimoides, 1.1 in M. discolensis, 1.9 in M. porosus and 1.6 in M. parviporosus

FIG.3
137 labial papillae and four cephalic setae, in different cycles. Cephalic setae corresponding
179 cuticularized, and with large teeth. However, M. campiensis sp. n. can be differentiated from 180 M. affinis by the cephalic setae length (46–50% of head diameter vs 37–43% in M. affinis).
Cephalic setae are not visible in the photos, thus it can be difficult for a reader to understand the description.
146 Seven pre-cloacal 147 papilliform supplements
Only 6 supplements are highlighted in the photo, but more than 6 are visible as described.
165 Presence of two glands, one on each ovary branch.
As for cs, glands may be difficult to check.

FIG.6
FIG.6 C cs visible but not clear
FIG.6 C and F show the same feature
FIG.6 G tail not visible

FIG.9
Cephalic setae not visible
Additional comments: Data and discussion do not contradict each other and there is sufficient data to support the conclusion. It can be improved by adding more references and clearer photos (with symbol legends, for male holotypes and female allotypes). Finally, fixing minor mistakes in the tables can be helpful in refining the work.


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: THREE NEW SPECIES OF MICROLAIMUS (NEMATODA: MICROLAIMIDAE) FROM THE SOUTH ATLANTIC
Review round: 1
Reviewer: 2

Output:
Basic reporting: General comments:
I congratulate the authors for making a valuable contribution to our knowledge of a complex genus, Microlaimus. My main reservation about the manuscript, which must be addressed by the authors, is that there must be more detailed information on the status of the genus, the known diversity (eg, how many species are there in the genus?), recent revisions and descriptions, and lists of species. The authors need to consult Kovalyev and Tchesunov (2005) and Leduc (2016), as well as the other authors they cite at the end of their Introduction (not just cite them but give a summary of their key findings, and also include an updated species list). At present, the authors do not convince me that they have made exhaustive comparisons with all valid species of the genus, because they do not demonstrate a familiarity with all of the relevant literature. Microlaimus is a large genus and one needs to be diligent in checking all of the literature to avoid describing species which are in fact already named. The authors should also realise that the Microlaimoidea are now classified within their own order, the Microlaimida (see Leduc et al. 2018 ZJLS), and not the Desmodorida.
I wish someone would review the genus and divide it into groups that would facilitate identification, and maybe devise a key. However I do not ask that the authors necessarily do this.

Specific comments:
Line 105: nematodes don’t have a head, they have a cephalic region.
Line 109: The appropriate taxonomic authority for Microlaimus dicolensis has been established to be Bussau, 1993, not Bussau & Vopel, 1999. See Holovachov (2020) Bionomina.
Line 110: a unispiral amphid has a circular shape by definition
Line 113: replace “sclerotized” by “cuticularized” here and throughout manuscript. Clarify whether this ring surrounds the whole buccal cavity (what do you mean by “at base of dorsal tooth?)
Line 116: males do not have an anus, they have a cloaca. Correct here and throughout manuscript.
Lines136-138: are the external labial sensilla papilla or setae? Choose one do not use both
139: posterior to, not “after”. Correct here and throughout manuscript
140: “with 12 folds at upper end” - do you mean “Cheilostoma with twelve longitudinal folds”?
Line 141: The correct term is ventrosublateral, not sublateral, see Coomans 1979 - A PROPOSAL FOR A MORE PRECISE TERMINOLOGY OF THE BODY REGIONS IN THE NEMATODE
Line 142: round number to 78% Cannot possibly be that precise. Same for line 139, 157, 161, 164 etc…
Line 153: You write “generally similar”, but is this a description of a single female? I don’t think that allotypes are used anymore, you need to check the Code.
Line 165: these don’t look like glands to me, more like sphincter muscles. Why would glands surround each genital branch? Their function cannot be to push the egg along!
Line 172:  “enlarged proximal end” – do you mean a capitulum?
Line 173: delete “strangler”, and see my comment above about muscle vs gland
Line 181: only one spicule? Usually they have two.
Line 244: what do you mean by “situated in same row”? clarify.
Line 248: provide dimensions of sperm for all spcies
Line 248: “with proximal portion cephalised” – you mean capitulum present?
You need to provide a better and larger drawing of the buccal cavity showing the unusual structure in M. alexandri (5 teeth)
Line 272-273: “The cuticular ring…” what do you mean by “same row” the ring needs to be better shown in drawing. Too small. You also need to show the longitudinal slits (openings of epidermal glands) in the drawings.
Line 401: testes, not testicles
Line 412: should be: “However, despite this lack of information…”
Table 2: no decimal points on the percentages. Here and throughout manuscript
Figure 1: the vas deferens near the spicules looks a bit like a posterior testis? On Figure 1A
Experimental design: see above
Validity of the findings: see above
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: THREE NEW SPECIES OF MICROLAIMUS (NEMATODA: MICROLAIMIDAE) FROM THE SOUTH ATLANTIC
Review round: 1
Reviewer: 3

Output:
Basic reporting: Three new species of Microlaimus (Nematoda:
Microlaimidae) from the South Atlantic (#53043)
Review Report

Thank you for the opportunity to review the manuscript on description of three Microlaimus species as entitled above.
This excellent work on taxonomy of marine nematode, it adds knowledge for biodiversity and improves the list of known species. The manuscript is well written and especially the line drawings are well done and clear. The tables are done with a lot of details that helps the reader compare the different species described here and with others. The descriptions of each species are elaborated and the diagnosis and differential diagnosis clearly set out.
I have however a few comments and 1 or 2 suggestions
1.	Line 33-34 there is indication of Microlaimus occurring along the continental shelf and slope, yet all the three species described were sampled from 25m. And so as far as the species described here are concerned, there is evidence of occurrence at continental shelf.
2.	Line 40-43: The statement suggests that Molgolaimus belongs to Microlaimidae why it belongs to Desmodoridae and more recent work by Leduc et al (2019) suggest puts it in Chromadorida.
3.	One weakness on the descriptions that runs throughout the three species is the focus of description and measurements on the male holotype alone and the female allotype. It would be best to consider all the individuals and thus report measurements as a range rather than single figure but keeping the male and female descriptions separate as they are currently is okay.
4.	Counter-check table referencing in the text. For instance, Line 134: Microlaimus campiensis should be Table 2 not 3 and this error runs through out the text.
5.	Line 135: It is easier to read the text and follow when actual measurements are mentioned, where necessary, before the relative sizes. For instance, ‘Maximum body diameter corresponding to 1.8x head diameter’, neither mbd, nor head diameter is given. I acknowledge all information is in the tables but some guiding data is necessary in the text.
6.	The structuring of sentences needs to be counter-checked. For instance, the sentence on 142-143 ‘ Nerve ring located at level of secretory-excretory pore, distance from anterior end equivalent to 60% of pharynx length’.  Line 164: ‘…… intestine, comprising 50% of body length’ sentence not clear.
7.	Line 174: I could not see the comb-like glands in the female ovary. It would be nice to show them either in the plate or line drawing more clearly. The description of the glands as ‘strangler’ needs to be elaborated.
8.	Line 221: Microlaimus alexandri sp. n. I have a problem with the number of individuals available for description of this species. With only one female and sexual dimorphism in the amphid size and location, one is left wondering whether the three individuals are really same species. The author needs to decide how to deal with that challenge of the numbers-delay publishing until enough material is found or describe only males and assume females were not found.
Thank you again for the opportunity

Agnes Muthumbi, University of Nairobi
Experimental design: Not applicable
Validity of the findings: Very useful results in the light of the need to document biodiversity in ecosystems
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: THREE NEW SPECIES OF MICROLAIMUS (NEMATODA: MICROLAIMIDAE) FROM THE SOUTH ATLANTIC
Review round: 2
Reviewer: 1

Output:
Basic reporting: English is fluent and professional
Clearer figures were added
Highlighted mistakes were fixed
Symbols were added to the respective figures
A conclusion chapter was added, but there are some typos:
•	429. Microlaimus in italics
•	429. species
•	423-427. Conclusion before acknowledgements
Experimental design: Now it is more detailed
Validity of the findings: Figures and table were corrected
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: THREE NEW SPECIES OF MICROLAIMUS (NEMATODA: MICROLAIMIDAE) FROM THE SOUTH ATLANTIC
Review round: 2
Reviewer: 2

Output:
Basic reporting: see attachment
Experimental design: see attachment
Validity of the findings: see attachment
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: PERFLUOROALKYL ACIDS IN SEDIMENT AND WATER SURROUNDING HISTORICAL FIRE TRAINING AREAS AT BARKSDALE AIR FORCE BASE
Review round: 1
Reviewer: 1

Output:
Basic reporting: Scientific writing needs to be improved
Literature should be updated
Data should be reanalysed, the presentation of averages of all chemicals removes all of the important information
Experimental design: Why was a temporal assessment chosen?
If this study was in support of an ecological assessment it would have been intersting to compare these results to what was found in the ecological assessment, as not all PFASs accumulate. An assessment of the contamination gradient with increasing distance from the point source could have been done.
Validity of the findings: There is nothing novel about the study in its current presentation
Additional comments: The study reports PFAA concentrations in sediment and surface water around an army base. It seems like a very extensive sampling campaign was conducted that could have been used for a really worthwhile assessment of contaminant distribution, however, the results presented are very limited, have major drawbacks in the analytical and add no new information.
Abstract: it promises more than what is behind the study. If the ecological assessment mentioned in the abstract will play no part in the entire manuscript it may just be better to leave it out. Where are the results of the spearman-rank correlation mentioned in the abstract? How do the authors come to the conclusion that local runoff from the use of commercial products (other than AFFF) contribute to the contamination observed?
Introduction: the introduction reads like a thesis, is very descriptive and cites publications from 10+ years ago. Some information contained in the introduction is not correct; i.e. PFOS is not the only PFAS listed in the Stockholm Convention, PFOA has also been listed and others are under review for inclusion. Up to date information on the use of AFFF in the US could have been included, as I am sure that has also changed in the last 10 years.
Materials and methods: the analytical standards (mass-labelled surrogates) selected are somewhat ill fitted, although it is impossible to tell which surrogate was used for quantification of which native. Usually matching standards are used, i.e. mass-labelled PFHxS for the analysis of native PFHxS and so on. Having at least some matching mass-labelled standards for such a limited standard suite of PFAAs is an absolute must. If no matching mass-labelled standards are available one would at least use those with the closest mass and the same headgroup as a surrogate.
Results and discussion: The results are presented as averages across the months and lump together all PFAAs analysed. Why was this done? All of the interesting information is lost that way and one is left with a bar chart of sediment averages that show no differences between the months with such high standard deviations. Sediments are also quite heterogeneous a variation is already expected between sampling events. The limitations of the sediment extraction method are more likely a results of the ill-fitted standards rather than the extraction method itself. The authors could have used the data it much better ways to describe the flow of chemicals away from the point source. For example changing ratios of chemicals, both in sediment and in water, which would also be related to the chemical properties.
Conclusions: a conclusions section usually reports the main findings of the study on hand. Here, the results are compared to a study from 2011.


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: PERFLUOROALKYL ACIDS IN SEDIMENT AND WATER SURROUNDING HISTORICAL FIRE TRAINING AREAS AT BARKSDALE AIR FORCE BASE
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript presents some interesting data on current PFAS concentrations at a historically AFFF-contaminated site. The manuscript is well-written and fairly clear, and the amount of background/context given was sufficient. However, some of the driving hypotheses are unclear. The manuscript could be improved by focusing in some more on what can be learned from this dataset - it seems the data point towards an AFFF point source coming from the FTAs, and that a comparison to older data from the site indicates that concentrations have not decreased. These are interesting findings, and the manuscript could be improved by focusing in on these points.

It is unclear what the driving hypothesis is for looking at average concentrations every month at all locations. We expect and observe significant variation site-to-site, so it might be better to focus on specific sites in Figures 1 and 3 to understand temporal variability.

Additionally, the results & discussion section could be improved by adding more depth - for example in lines 283-288, it is mentioned that a previous study found higher concentrations than those found in this study. This observation is mentioned briefly but no explanation is given as to what it means or why it might be significant. Delving into a comparison of previous measurements versus those here would be really interesting.

Other specific comments:

Line 83: missing word between "the" and "and"
Lines 100-101: here "PFASs" is used whereas before the authors used "PFAS" as the plural form - please check through the manuscript and make sure you are consistent with how you're handling PFAS acronyms.
Lines 224-225: here it is stated that concentrations at RLs were quantifiable in Late May, but in Figure 2 it is stated that all PFAS were <LOD at RLs.
Line 282: A correlation is observed between chain-length and water concentration. I think the discussion of this needs to be expanded upon for it to really be a useful observation. Also, the authors should make it clear that while there are many data points from different samples, there are few compounds (so few chain-lengths) being used for the regression. What's the meaning of this regression? Could it be due to the original AFFF composition?
Experimental design: Methods:

For the most part, accepted methods were used for sample preparation and analysis. However, I have some concerns about methodology, particularly related to surrogate recoveries, and whether variation in sample-to-sample recovery could be affecting the observations made.

General comments:

As I'm sure the authors are aware, the low SURR concentrations for sediment extraction are of concern, as they are outside of what would usually be considered acceptable. Because the IS's used for quantitation do not correspond to the target analytes, that makes it more difficult to understand how robust the data is...

One way to address this issue to some extent and increase confidence in the data would be to describe the SURR recovery variability. While for the most part, the authors appear to have been quite diligent about QC, I saw no mention of collection of replicates, or the variability in replicate samples. This would be really helpful in understanding if seasonal changes observed are actually significant. Additionally, if low SURR recoveries are consistent, it increases confidence in observed trends.

Specific comments:

Lines 110-120: Typically when quantifying PFAS, you want to be very careful to quantify using the corresponding labeled compound whenever possible to avoid matrix effects and differences in ionizability that are difficult to predict. I found it very unusual that sulfonamide standards were used here to quantify PFAAs, and I think this should be addressed in the manuscript. Furthermore, the authors should specify which IS was used to quantify which compound. Choice of IS can affect quantitation.
Validity of the findings: The manuscript is transparent in providing raw data. Additional confidence could be gained if a presentation of replicates was given, and if IS quantitation was explained further and uncertainties due to non-ideal standards being used were explicitly described.

The conclusions would be strengthened if more detail was added to the discussion, particularly in relation to temporal comparisons (both seasonal and comparison to previous study).
Additional comments: Thank you for your work on this interesting study. You have produced some data that will be of interest to others studying AFFF-impacted sites. My comments on how to further improve the manuscript are detailed in the previous sections.


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: PERFLUOROALKYL ACIDS IN SEDIMENT AND WATER SURROUNDING HISTORICAL FIRE TRAINING AREAS AT BARKSDALE AIR FORCE BASE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript has been improved based on my initial round of review and I have no further suggested edits.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: LONG-TERM CONSUMPTION OF ARTIFICIAL SWEETENERS DOES NOT AFFECT CARDIOVASCULAR HEALTH AND SURVIVAL IN RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: line 180-181: should be referenced properly.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The topic is interesting and the data unique. The authors present interesting data about the Long-term consumption of Artificial Sweeteners and how it does not affect cardiovascular health and survival in Rats especially with the current pandemic that affected research I find that it is a very interesting study with great results.
The manuscript is well written, has important message, and should be of great interest to the readers.


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

Input:
Title: LONG-TERM CONSUMPTION OF ARTIFICIAL SWEETENERS DOES NOT AFFECT CARDIOVASCULAR HEALTH AND SURVIVAL IN RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is a straightforward paper.  However, much of the scientific literature on artificial sweeteners has not been included.
Experimental design: The word "batch" should be changed to "group" throughout the manuscript.  It is troublesome that fluid intake was not measured daily.  This is necessary to make conclusions about dosages.
Validity of the findings: It is hard to determine the validity of findings due to the estimates of sweetened water intake as noted above.  There were also issues of monitoring during the Covid crisis.  However, the findings on electrophysiological alterations is a very important observation if correct.  The prevalence of atrial fibrillation is increasing in the human population and should be addressed in the discussion.
Additional comments: A correlation between AFIB and artificial sweetener use would be helpful here.


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

Input:
Title: LONG-TERM CONSUMPTION OF ARTIFICIAL SWEETENERS DOES NOT AFFECT CARDIOVASCULAR HEALTH AND SURVIVAL IN RATS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The report is well structured; the language is clear and concise. The introduction provides a good context that leads to the research question. In line 58 the association of cardiovascular diseases and sex are considered as relevant background, but the possible influence of sex on cardiovascular diseases related to the sweetener used is not retaken. Considering the sample size of each group, it is possible to carry out a statistical analysis that considers this factor. Figure captions could explain better their content.
Experimental design: The study’s experimental design is ambiguous, it is not clear whether the authors seek to explore the differences between the sexes. For example, it is not clear if the comparison of weight and water consumption between groups was made by sex, that is, a comparison between groups for females and another for males, or if a comparison was made considering only the treatments (line 171).

Additionally, given that weight and water intake were collected repeatedly, it is convenient to perform an analysis that considers the repeated measures (for example, an ANOVA of repeated measures if the data meet the criteria of sphericity). If possible, I strongly suggest performing a statistical analysis that considers the effect of sex, treatment, and the repeated measures.
It is not clear whether the survival analysis was performed with both sexes or if each sex was analyzed separately. The results mention that survival was higher in females, but the authors do not mention the test used to compare the preference curves (line 186).
I suggest adding the mean of the groups ± SD, the values of F (assuming that an ANOVA test was performed) and the value of p in blood pressure and pulse wave velocity results(line 190). In the same sense, the means for the AVCT and ERP measures, the value of the test used, and the adjusted p values of the multiple comparisons (line 200) should be added, as well as the means of blood lipids and glucose levels, the test and p-values (line 212). There is no reference chart for blood glucose values (line 214).
Validity of the findings: The discussion sounds more like a review. I suggest that the authors compare how their findings fit in with the existing literature. A better discussion considering the differences between sexes could be written. I list some articles that report that the effect sweet foods is more pronounced in adult females (Valenstain, 1967, J Comp Physiol Psychol, 63 : 429-433; Sclafani et al., 2010, Chem Senses, 35: 433-443; Marshall et al., 2017, PLoS One, 12 [7]: e0180907).
Authors should mention that the sweeteners used in their research is a commercial mixture. Findings in humans suggest that the intake of beverages containing a mixture of sucralose and maltodextrin have different effects on the AUC of insulin related to the intake of beverages containing only sucralose or maltodextrin (Dalenberg et al., 2020, Cell Metab, 31: 493-502.e7). Therefore, I suggest the discussion should include literature with the effects of the specific sweeteners used in the study.
I consider that the most important result to discuss is that the prolonged use of splenda had an effect on left ventricular hemodynamics (line 225). In this regard, Risdon et al. (2021, Nutr Metab Cardiovasc Dis, 30: 843-846) have reported that the intake of a mixture of acesulfame and sucralose has an effect on the endothelial vascular function of adult rats. Certainly, the incorporation of related literature will help enrich the discussion.
Additional comments: It is interesting research with considerable data, but it lacks a concise approach to the interpretation of the results. A better discussion can give more validity to the work.


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: FULL-LENGTH TRANSCRIPTOME ANALYSIS OF ADIANTUM FLABELLULATUM GAMETOPHYTE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The language of this manuscript needs further enhancement. Problem statement in the abstract and Introduction is unclear. Simply stating “The study of gene expression is rarely reported” is not a strong enough reason to conduct full-length transcriptome analysis. I suggest the authors to highlight the potential impact of their study.
Experimental design: Experimental Design
Some parts of the experimental design are unclear. For example:
1.	Line 80-82: How many gametophytes were used for RNA extraction?
2.	Line 86: were the gametophytes pooled according to illuminations?
Others are quite standard.
Validity of the findings: no comment
Additional comments: The study is a fundamental descriptive study. The authors should have incorporated treatment comparison, e.g. comparison of gametophytes at different illuminations, or different stages, in order to derive more meaningful results. The current manuscript lacks importance and is merely a basic study to provide a reference baseline for future research.


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

Input:
Title: FULL-LENGTH TRANSCRIPTOME ANALYSIS OF ADIANTUM FLABELLULATUM GAMETOPHYTE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Comments on language and revision: The English language should be improved to ensure that an international audience can clearly understand your text.  I suggest you have a colleague who is proficient in English and familiar with the subject matter review your manuscript, or contact a professional editing service. The sections that need particular attention is the abstract and introduction.

Sufficient background and context has been provided in the literature review to understand the aims and rationale of the study.

The study is logically structured and figures and tables are appropriate.

Data: I have looked at the Bioproject and accession and is a bit worried about the file size reported and the data reported in the study. The Bioproject indicate that 113 Mb of 463 542 116 bases has been submitted, while the project indicate that 69 Gb of bases were generated. Perhaps just confirm that these are correlated since I would have expected a few Gb of data. Is this just compressed?
Experimental design: The study lies within the aims and scope of the journal.
The research proposed fills a knowledge gap and the aim of the study is certainly meaningful even if no specific research question was posed.
The experimental procedures are standard for this kind of transcriptome sequencing study and may therefore be considered rigorous.

Methods are overall described with enough detail to replicate although specific references to some of the general methodology used has not been included.For example:
Line 86: Concentration of RNA obtained and methodology for cDNA library construction.
Line 87-89: Indicate software used for clustering, correction, consensus read and redundancy removal.
Line 93: A reference for Transdecoder. In this regard, provide software version numbers for all software used.
Line 95: Provide references for all relevant databases used.
Line 104-108: Provide references for software and databases.
Validity of the findings: The results are standard for this type of study and overall robust and statistically sound.
The conclusions are well stated and founded and support the results.
Additional comments: Figure 1: Y-axis legend should be number.


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: FULL-LENGTH TRANSCRIPTOME ANALYSIS OF ADIANTUM FLABELLULATUM GAMETOPHYTE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The overall flow and structure of the manuscript are clear and concise. Tables and figures look good.
Experimental design: This manuscript provides the full-length transcriptome data of A. flabellulatum. Albeit the current manuscript reporting is merely descriptive in nature, this work serves as important baseline comparison reference for future transcriptomic studies of this species. The full-length transcriptome sequencing dan data analyses are rather standard. Additionally, the authors validated the transcriptome data using qPCR, which provided further support to the study. I noticed that there is an inconsistency in the Method, i.e., line 85-88. Did the authors pooled the three replicates of each illumination into one (line 86), or did they extract the RNA of each replicate, as noted in line 92.
Validity of the findings: no comment.
Additional comments: Please add more to the first paragraph of the Introduction. In addition, kindly rephrase the two sentences (line 44-47), 'play an important role' can be found in both first and second sentences.


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

Input:
Title: FULL-LENGTH TRANSCRIPTOME ANALYSIS OF ADIANTUM FLABELLULATUM GAMETOPHYTE
Review round: 2
Reviewer: 2

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


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

Input:
Title: DISTINCT METABOLIC PROFILING IS CORRELATED WITH BISEXUAL FLOWERS FORMATION RESULTING FROM EXOGENOUS ETHEPHON INDUCTION IN MELON (CUCUMIS MELO L.)
Review round: 1
Reviewer: 1

Output:
Basic reporting: General comments:

In the submitted manuscript, the authors described the metabolimc analysis in melon and more particulary in shoot apex of main stem from ethephon treatment and control plants to investigate the different metabolites contributing the occurrence of bisexual flowers.

The authors conducted extensively metabolomic analyses, combined with the experimental validation of phenotypes after etephone treatment. The strong points of the manuscript is a very interesting topic regarding to plant reproduction. The description of pathways are very interesting especially with regard to hormone signaling pathway connections.
Experimental design: The current guidelines for metabolomic analysis are that experiments using metabolome profiling data to describe changes should contain at leat three the biological replicates (preferably is more) and also three technical replicates. Herein authors have only biological replicates, which are bulked. According to the standard protocol, it  must contain three biological and three technical replicates. Herein there is lack of this information and thus it seems the  analysis are not properly replicated. In materials and methods section – Materials and methods section - p.10 line 139 Five shoot apexes were mixed as one biological replicate and three biological replicates were performed for ET and CK. Please look at the technical quiadance for recommendations for metabolomic profiling data: Fernie AR, Aharoni A, Willmitzer L, et al. Recommendations for reporting metabolite data. Plant Cell. 2011;23(7):2477-2482. doi:10.1105/tpc.111.08627
Validity of the findings: Has statistical reference been made to all identified metabolites? An example is the Bonferoni correction, which adjusts the observed significance level to the fact that many comparisons are made.

The diagram presented by the authors at the end of the article and the conclusions do not have much support in the results described.
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: DISTINCT METABOLIC PROFILING IS CORRELATED WITH BISEXUAL FLOWERS FORMATION RESULTING FROM EXOGENOUS ETHEPHON INDUCTION IN MELON (CUCUMIS MELO L.)
Review round: 1
Reviewer: 2

Output:
Basic reporting: This is seems to be good article related to gene expression for flower development in lateral and main axis in melon
Experimental design: Appreciate and carried out properly
Validity of the findings: Findings are seems to new and novel.
Additional comments: Probably it is a new article for sex expression in relation to main axis and lateral branches in melon.


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: DISTINCT METABOLIC PROFILING IS CORRELATED WITH BISEXUAL FLOWERS FORMATION RESULTING FROM EXOGENOUS ETHEPHON INDUCTION IN MELON (CUCUMIS MELO L.)
Review round: 1
Reviewer: 3

Output:
Basic reporting: This paper mainly reports metabolic profiling contributes to the formation of bisexual flowers on main stem. By measuring metabolomics and endogenous hormone between main stem with or without ethephon treatment groups, the authors finally identified three significantly enriched KEGG terms participating in the occurrence of bisexual flowers in melon. The endogenous hormones SA and IAA were up regulated by ethephon, might be due to amino acids and organic acids up regulated.
However, there are some major concerns. The most important one is that these are only speculations how substances regulate the formation of hermaphrodite flowers, lacking profound evidence at the molecular level. Ge et al. (2020) have shown the transcriptome data of main stem with and without ethephon treatment. I recommend the authors use the strategy of metabolome and transcriptome joint analysis to construct co-expression network that provides comprehensive information on the dynamics of major metabolites and the transcriptome profiles that underlie them during melon bisexual flower development. By analyzing the co-expression network, the authors could explain metabolites variation from the level of gene expression in bisexual flowers development. It will be more powerful to strengthen the key points that ethylene regulates bisexual flowers development by crosstalking with SA and IAA through regulating some amino acids and organic acids.
Experimental design: This paper provides a potential perspective that metabolites play important role in regulating the bisexual flower development.
However, the title “Distinct metabolic profiling contributes to the formation difference of bisexual flowers between main stem and lateral branches in melon (Cucumis melo L.)” of this article is not appropriate, because in the author's experimental design, only the main stem was treated with ethylene, and the lateral stem group was not set. Both main stem treated with ethylene and lateral stem produce bisexual flowers, and the overlap factors between them should be the causal factors of bisexual flowers formation. Thus, according to this title, lateral stem should be used as another experimental group, except for main stem and main stem treated with ethylene.
Validity of the findings: 2.	In Figure 5, it seems that the differences of contents of metabolites such as shikimic acid, L-tryptophan, L-phenylalanine was not significant, and the raw data should be provided to confirm the conclusion. Also, the number of biological duplicates should be added.
Additional comments: 3.	In line 94, there is a spelling mistake that “stamina” should be replaced by “stamens”.


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: DISTINCT METABOLIC PROFILING IS CORRELATED WITH BISEXUAL FLOWERS FORMATION RESULTING FROM EXOGENOUS ETHEPHON INDUCTION IN MELON (CUCUMIS MELO L.)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is clear and English language seems to be good in thorough out the manuscript. Structure of the article also seems to be as per the standard of this journal.
Experimental design: Sufficient
Validity of the findings: seems to be novel and certainly have academic significant.
Additional comments: This manuscript has been well written and corrections have been incorporated,


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: ASSOCIATIONS BETWEEN THE RS5498 (A > G) AND RS281432 (C > G) POLYMORPHISMS OF THE ICAM1 GENE AND ATHEROSCLEROTIC CARDIOVASCULAR DISEASE RISK, INCLUDING HYPERCHOLESTEROLEMIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: Here, they reported that the G allele in ICAM1 rs5498 is linked to increased ICAM1 mRNA expression, LDL-C concentration, and RHR, which might also lead to increased ASCVD risk and HCL in Thai adults.
Experimental design: ICAM1 SNPs Genotyping was performed in normal controls and HCL individuals assessed by clinical characteristics.
Validity of the findings: This prevalence study analyzing data from 278 participants in Thailand and showed the relationships between the ICAM1 rs5498 (A>G) polymorphism and hypercholesterolemia (HCL) and the atherosclerotic cardiovascular disease (ASCVD) risk factors in them.
Additional comments: (1)	Although the data shown in this study suggested that ICAM1 rs5498 (A>G) might be linked to HCL in ASCVD, its effect on increased ICAM1 mRNA level was not significant (Table 1 and Table 3). Only in Fig. 2, normal control with wild type AA showed the significantly less expression of ICAM mRNA than HCL group with variant (AG+GG).
(2)	In this study, it was not clear why the G allele in ICAM1 rs5498 is closely linked to increased the risk for ASCVD and HCL in participants.
(3)	Does increased ICAM1 expression lead to the increased heart rate or increased sympathetic activation?


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: ASSOCIATIONS BETWEEN THE RS5498 (A > G) AND RS281432 (C > G) POLYMORPHISMS OF THE ICAM1 GENE AND ATHEROSCLEROTIC CARDIOVASCULAR DISEASE RISK, INCLUDING HYPERCHOLESTEROLEMIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Wechjakwen and colleagues investigate the role of two polymorphisms of ICAM gene on hypercholesterolemia and atherosclerotic cardiovascular disease. Despite the research topic being relevant, this study has some flaws and it is not completely clear what new message the authors want to bring in comparison with the other studies of the field.
Hence the authors should make modifications to make the manuscript clearer.

1. In the abstract the risk factors that are going to be taken in consideration should be introduced, to have an overview of the analyses performed.

2. Study design and participants: How subjects where recruited?
3. Study design and participants: How enviromental variables were assessed?
4. Line 115: normal control group should be defined as control only. And wild-type is not completely appropriate when we are talking about genotype. It's reference allele the correct definition.
5. Table 4. a. the same analysis should be shown also for rs281432. b. P-value for Chi-squared test should be reported for the difference between low-risk and high risk c. c.The part of mRNA expression should be moved separately. d. Is there some reason why blood pressore was not included?
6.Discussion. The authors should be more cautios in saying that they demonstrate the association of the snp with ASCVD. They just reported the association with some of the risk factor separately.
Experimental design: The techniques used were correct.
Validity of the findings: There would need further investigation including inflammatory markers, to validate the analyses.
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: ASSOCIATIONS BETWEEN THE RS5498 (A > G) AND RS281432 (C > G) POLYMORPHISMS OF THE ICAM1 GENE AND ATHEROSCLEROTIC CARDIOVASCULAR DISEASE RISK, INCLUDING HYPERCHOLESTEROLEMIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors provided improvements to the manuscript, for all the issues proposed in the previous review process.
Experimental design: The authors provided improved the manuscript for all the issues proposed in the previous review process.
Validity of the findings: The authors provided improved the manuscript for all the issues proposed in the previous review process.
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: COMPARISON OF THE COMPOSITION AND FUNCTION OF GUT MICROBES BETWEEN ADULT AND JUVENILE CIPANGOPALUDINA CHINENSIS IN THE RICE SNAIL SYSTEM
Review round: 1
Reviewer: 1

Output:
Basic reporting: Manuscript :
Comparison of the composition and function of gut microbes between adult and juvenile Cipangopaludina chinensis in the rice snail system

The manuscript described the differences in gut microbiota complexity in Juvenile, adult, and sediment of Cipangopaludina chinensis. The work is novel as characterization of mollusk microbiota did not clarify or extensively studied as other aquatic fish species. I recommend the acceptance of the manuscript but after minor revision as shown below;
In the introduction more detailed about the anti-cancer effects of Cipangopaludina chinensis could increase the scientific value of the manuscript.
 The authors stated that”Therefore, C. chinensis can not only be used as food, but also have great potential in human disease prevention and treatment” In relation to this more information about the nutritional values and ingredients of this mollusk is needed. Additionally, the hepatoprotective effects should be mentioned.
In the discussion the authors should highlight the abundance of healthy bacteria (if possible) among the identified gut microbiota either in Juvenile or adults.
Results are clearly presented and the references are updated
Experimental design: The experimental work is well designed, clear, and reproducible
Validity of the findings: Up to the economical problems especially in third world and poor countries working on snails as source of nutritional proteins could be of great help. Based on the importance of gut-organ axis and its relationship to health and disease, characterization of C. chinensis  microbiota could help the industrial contribution to increase the abundance of   C chinensis probiotics as nutritional intervention.
Additional comments: The manuscript is well written and the interpretation is satisfactory


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: COMPARISON OF THE COMPOSITION AND FUNCTION OF GUT MICROBES BETWEEN ADULT AND JUVENILE CIPANGOPALUDINA CHINENSIS IN THE RICE SNAIL SYSTEM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is good written. For further improvement, few suggestions are recommended for revision as follows and as shown in the annotated manuscript.
Experimental design: 1.	The introduction section needs more details regarding feasibility of the sequencing method for this study, especially why 16SrRNA is used.
2.	 In lines, 53-55 and 60-61, rephrasing is required with correct language expression.
3.	In line 99, “Bioinformatics and community comparisons” should be expanded.
Validity of the findings: In the reference section (lines, 303-305), names of journal should be appropriate as they are shown originally.
Additional comments: N/A