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

Input:
Title: IDENTIFICATION OF DIAGNOSTIC GENES AND VITAL MICRORNAS INVOLVED IN RHEUMATOID ARTHRITIS: BASED ON DATA MINING AND EXPERIMENTAL VERIFICATION
Review round: 1
Reviewer: 1

Output:
Basic reporting: English language is a concern throughout the manuscript. Another point is consistency of writing and use of terminology.
References aren't appropriately used.
Introduction is  not well connected.
Experimental design: No original research.
No gene enrichment analysis was performed.
Research question was not well defined.
There is no rigorous investigation performed.
Validity of the findings: No novelty, this type of reports already published. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6657850/).
Results were not interpreted and explanatory.
This is just use of bioinformatic tools study. Author only used qRT PCR to validate the genes but not western blots for their protein products.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF DIAGNOSTIC GENES AND VITAL MICRORNAS INVOLVED IN RHEUMATOID ARTHRITIS: BASED ON DATA MINING AND EXPERIMENTAL VERIFICATION
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript titled "Identification of diagnostic genes and vital
microRNAs involved in rheumatoid arthritis: based on
data mining and experimental verification" has been written well. There are very few grammatical errors in the manuscript with sufficient literature review provided. The data analysis and representation is adequate. The authors have missed to add the findings of a) Clin Exp Rheumatol. Jul-Aug 2017;35(4):586-592. Epub 2017 Jan 27.; b) Zhu X, Wu L, Mo X, et al. Identification of PBMC-expressed miRNAs for rheumatoid arthritis. Epigenetics. 2020;15(4):386-397. doi:10.1080/15592294.2019.1676613. The authors should consider including them in the discussion.
Experimental design: The experimental design is well planned to address the hypothesis with well defined objectives. There is adequate information for the repetition of the experiment by others.
Validity of the findings: The findings are novel for the biomarkers although there are reports on the miRNAs involved in RA patients. The conclusions are well supported with the findings of the study. The authors should validate a few of the miRNAs for functional role in regulating the inflammatory genes mentioned in the study as a functional validation of the study.
The authors have mentioend that the highly altered pathways belong to inflammation and cell-cell adhesion but they have not validated any gene for the cell-cell adhesion pathway or looked into its miRNA regulation. Why?
Additional comments: The authors should rectify the grammatical errors and space between the words in 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: IDENTIFICATION OF DIAGNOSTIC GENES AND VITAL MICRORNAS INVOLVED IN RHEUMATOID ARTHRITIS: BASED ON DATA MINING AND EXPERIMENTAL VERIFICATION
Review round: 2
Reviewer: 1

Output:
Basic reporting: Clear and unambiguous and language is appropriate.
Sufficient field background and references
Figures and structured articles
Experimental design: Articles fit in Aims and scope of the Journal
Research question is improved in revised manuscript.
Investigation in details.
Methods described.
Validity of the findings: Findings are well defined. All data have been provided. Conclusions are well stated.
Additional comments: Author address my concerns.


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

Input:
Title: IDENTIFICATION OF DIAGNOSTIC GENES AND VITAL MICRORNAS INVOLVED IN RHEUMATOID ARTHRITIS: BASED ON DATA MINING AND EXPERIMENTAL VERIFICATION
Review round: 2
Reviewer: 2

Output:
Basic reporting: The language can be made better
Experimental design: The question is well defined. The GSEA has made this manuscript better.
Validity of the findings: Conclusions could have been better stated.
Additional comments: This reviewer is of the opinion that the authors have done a good job with the rebuttal. However, though not fully agreeing with the other reviewer's comments about the research not being entirely novel, I feel that the conclusions of this manuscript can be made much better.
1) The strength of this manuscript is in the microRNAs prediction. Hence, I would suggest the authors to expand greatly on the  ROC curve prediction and microRNA validation.
2) I would highly recommend running a few Western blots for protein validation
3) Please expand the discussion to put more focus on the microRNA validation and the results from the Western Blots (if run) from point 2


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

Input:
Title: IDENTIFICATION OF DIAGNOSTIC GENES AND VITAL MICRORNAS INVOLVED IN RHEUMATOID ARTHRITIS: BASED ON DATA MINING AND EXPERIMENTAL VERIFICATION
Review round: 3
Reviewer: 1

Output:
Basic reporting: Good
Experimental design: Good
Validity of the findings: Novel. Good
Additional comments: Congratulations! The authorshave satisfactorily answered all my concerns. The manuscript is very good and should be published.


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

Input:
Title: EGFR MUTATION STATUS YIELD FROM BRONCHOALVEOLAR LAVAGE IN PATIENTS WITH PRIMARY PULMONARY ADENOCARCINOMA COMPARED TO A VENOUS BLOOD SAMPLE AND TISSUE BIOPSY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Nikolay et al. reported the testing results from bronchoalveolar lavage potentially be used for EGFR mutation status testing. The conclusion of this manuscript is liquid biopsy sample for EGFR status from BAL is superior to a venous blood sample. For this work, I mainly have some questions and comments as follows.
Experimental design: 1. In this study, why did the authors choose serum samples instead of plasma samples to analyze the EGFR mutation status? The serum sample contains a good deal of cfDNA, it affect the testing rate.
2. Which platform their used to test the EGFR status for plasma and BAL samples? They should describe it in the method section.
Validity of the findings: No comments.
Additional comments: The authors should reorganize the study design and data, and discuss the advantages and disadvantages of the three approaches for EGFR status testing, rather than simply draw a conclusion that the BAL testing is superior to the blood testing.


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: EGFR MUTATION STATUS YIELD FROM BRONCHOALVEOLAR LAVAGE IN PATIENTS WITH PRIMARY PULMONARY ADENOCARCINOMA COMPARED TO A VENOUS BLOOD SAMPLE AND TISSUE BIOPSY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is written in English language that is unambiguous and rather clear. Please consider a minor revision.
Regarding, the background described by authors introduces well into the context of the study. The urge of using liquid biopsies instead or as a surrogate of tissue biopsies is well justified. I am however missing a one-two sentence link to the use of BAL specimen. In addition, I am missing a mention of reports on the current knowledge. Regarding EGFR detection in blood and BAL – especially that this study is not novel in that sense. More emphasis should be put on the fact that this is PCR not NGS method.
Most of the references cited for this study is relevant. Reference [7] cited is a little obsolete: the methods of EGFR detection cannot be compared directly, moreover there is also no clarity regarding the material used (ctDNa from BAL supernatants or whole BAL sample). Once clarified the issue from pt. 1, please go back to the references and discuss accordingly.
Please revise Reference [14] for its details that appear in PubMed. Found only this: Carstensen T, Schmidt B, Engel E, Jandrig B, Witt C, Fleischhacker M. Detection of cell-free DNA in bronchial lavage fluid supernatants of patients with lung cancer. Ann N Y Acad Sci. 2004 Jun;1022:202-10. doi: 10.1196/annals.1318.031. PMID: 15251961.
Experimental design: The most important issue to comment is that it is not entirely clear if the authors worked with ctDNA from BAL (supernatant) or DNA isolated from the cellular debris. Based on the kit used (Circula, I assume the first. If the BAL was not processed via centrifugation (if was – please provide info about the speed, time etc) – we can hardly agree on the fact that it was ctDNA – it could derive from the cells that are present in the BAL. Although this detail does not impair the utility of BAL as a diagnostic tool, this point merits certain clarification.
In Materials and Methods please add the provider of all the kits. In addition, for more clarity I would suggest to move the paragraph regarding Ethical Committee where patient recruitment. I suggest dividing this part into 1) Patients 2) Samples collection and processing 3) DNA extraction and genotyping.

The study is missing table with patient characteristics.
Reg. Table 2 I suggest changing the Legend. Either mark with “*” only the novel confirmed mutations (pts 8 & 12) or add another mark for these additional mutations detected in blood/BAL. This is an important information and gets lost in the Table.
Validity of the findings: This study is not novel in the field. It does provide, however, meaningful confirmation of the utility of BAL for EGFR detection.

Would be very interesting to see the possibility of detection of a panel of mutations in these samples, however it is understandable that the clinical utility of such study must come in line. Namely, if there is no centrally refunded therapies targeted in other drivers that EGFR in Bulgaria, probably has not much sense to be performed today, but could serve as a good premise to include other targeted therapies into clinical practice. Maybe you could speculate on that in a one-sentence more of the discussion.
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: EGFR MUTATION STATUS YIELD FROM BRONCHOALVEOLAR LAVAGE IN PATIENTS WITH PRIMARY PULMONARY ADENOCARCINOMA COMPARED TO A VENOUS BLOOD SAMPLE AND TISSUE BIOPSY
Review round: 1
Reviewer: 3

Output:
Basic reporting: Basic Reporting:
In general, the manuscript is well written considering the authors are from Bulgaria.  Context is easily understandable, but would benefit from a professional medical writer fluent in English to clean up the paper. The text is technically correct.
The Introduction and background are sufficient for the content of the paper. The comment on lines 50-51 regarding the difficulty of obtaining sufficient tumor tissue needs to be referenced such as in the paper by Wu et al (Lung Cancer, 2018, Vol 126, pp 1-8). On line 54, it is unclear how reference 3 relates to complications of a tumor biopsy, could a better reference be found?
The structure of the article is consistent with accepted norms. There are only two tables which are relevant to the content of the article. Raw data in the form of an excel file of EGFR mutations which is germane to the content of the article. There should be a demographic table on the patients in the study, including  age, sex, performance status if available, smoking status and stage.
The article appears to be self-contained and a complete body of work and not subdivided from a larger body of work.
Experimental design: Experimental design
The article is within the scope of the journal related to the medical sciences.  The research question is adequately defined and meaningful for patients with lung cancer.  The authors define a medical need and the study contributes to bridging the medical need. The investigation appeared to be conducted in accordance with accepted norms. On lines 94-95 the authors state that the protocol was approved by an ethics committee and informed consent was obtained.  However, it is not clear when informed consent was obtained.  Was it obtained prior to the initial bronchoscopy or after lung adenocarcinoma was confirmed (line 71)?  Additionally, how was the lung cancer confirmed? Was it by a tissue biopsy, cytology or something else? What was the gold standard used in this study?
The flexible bronchoscopy is described in reasonable detail (lines 73-79).  However, the broncho-alveolar lavage (BAL) (line 79) is not well-described and should be expanded upon. What type of fluid was used and how much?  Was there a standard recovery?  If the technique used is detailed elsewhere, it should be referenced.  However, information on the amount of lavage fluid recovered would be interesting to include in the article.
The authors used the therascreen® EGFR Plasma RGQ PCR Kit for testing BAL specimens. This polymerase chain reaction (PCR) assay has been validated in plasma (therascreen® EGFR Plasma RGQ PCR Kit Handbook, January 2019) and not in lavage samples.  Did the authors conduct their own laboratory validation of the assay, as Ct cutoffs for PCR assays usually vary be sample type. Please describe any validation performed.

The authors list 112 patients with a “tumor” (line 99) but do not specify what this means.  Were they malignant?  Were they adenocarcinomas?  They should expand here.  A table with the full 140 bronchoscopies listing sex, age, maybe smoking status, tumor type (adenocarcinoma, squamous cell, etc.)  It is also stated that only 23.2% (26) of the 112 tumors were adenocarcinomas which seems unusual without knowing what the others were.  Among 112 primary lung malignancies, one would expect at least 60% to be adenocarcinomas or non-squamous cancers.  Please explain this.

The authors state that among the 26 adenocarcinomas, there were 13 that were EGFR positive and 13 that were EGFR negative.  This proportion (50%) is too high for a European population where one would expect an EGFR positivity rate of 10-15% (Lancet Oncology, 2012, Vol 13, pp239-46) as was seen in the EURTAC trial (14%). Please reconcile this.  Were there actually more than 26 adenocarcinomas and 13 were EGFR positive and the 13 EGFR negative patients were chosen from the remaining adenocarcinomas?  If so, how were these 13 chosen?
Validity of the findings: Validity of findings.

The authors present their data in a straightforward manner using the tissue biopsy EGFR results as the truth set which seems appropriate.   Underlying data was provided using a commercially available EGFR assay.

The conclusions need some editing.  The comment (line124) needs context.  EGFR testing by BAL may be better (not superior) to plasma EGFR testing in certain situations.  The authors failed to emphasize the most interesting finding in that 23/26 of the patients had visible intra-lumenal tumor. They should include how many of the EGFR positive samples had visible.  In this context, EGFR testing may be better than plasma EGFR testing in patients with visible intralumenal tumors.

In the same first paragraph of the discussion, the authors state that the BAL results provided additional mutation data in cases 6,8 and 12.  Was there any consideration to do orthogonal testing with NGS to confirm discordant results?

In the second paragraph of the discussion (lines 131-139), the authors discuss the low sensitivity of plasma EGFR testing compared to histological samples and do not recommend plasma testing.  Commercially available PCR based, plasma EGFR assays (cobas EGFR Mutation Test and the therascreen EGFR assay) clearly state that plasma testing is best suited for those patients in whom a tissue sample is not available.  Furthermore, the sensitivity and specificity of a plasma test is dependent on which EGFR mutation is tested and also the patient’s tumor burden. Thress et al (Lung Cancer, 2015, vol 90, pp 509-15) show much better sensitivity and specificity for exon 19 deletions and L858R mutations for both the cobas and therascreen assays than is stated in the paper (line 137).  Wu et al (Lung Cancer, 2018, Vol 126, pp 1-8), showed that the rate of plasma EGFR positivity was correlated with tumor burden.  The authors should expand on this paragraph for better context.  As they have the tumor diameters for 17 of the 26 samples, did they look at those patients who were plasma positive based on the size of the tumor?  It is fair to say that plasma EGFR testing is less sensitive, but as the thrust of this paper is about EGFR positivity in BAL, the statement regarding not recommending plasma EGFR testing (line 137-138) should be deleted.  Just stating that plasma testing is less sensitive than tissue testing should be sufficient here.

The authors state that BAL is a semi-invasive method with a lower risk for the patient (line 142).  Lower risk than what?  Tissue sampling?  If so, please provide references that support this contention.

In lines 163-164, the authors should state that BAL can be an alternative method to establish EGFR mutational status, especially in patients with visible intralumenal tumor.  Again, the most interesting finding was that 90% of patients had visible tumor and I would emphasize that consideration of BAL for EGFR testing should be considered in this situation.

The conclusion is over-stated.  BAL may be better than plasma EGFR testing in certain situations and provide valuable information for the treating physician.
Additional comments: General comments:

The information presented in this paper is interesting and adds to a relatively small body of work on using BAL to perform EGFR testing.  Most of the comments should be easy to adjudicate.  I appreciate the opportunity to evaluate 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: EGFR MUTATION STATUS YIELD FROM BRONCHOALVEOLAR LAVAGE IN PATIENTS WITH PRIMARY PULMONARY ADENOCARCINOMA COMPARED TO A VENOUS BLOOD SAMPLE AND TISSUE BIOPSY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Minor revision of English has been performed. Authors have referred to comments regarding the references accordingly.
I have no new comments to make on that section.
Experimental design: I am mostly satisfied with Authors' comments and changes.

Please only make sure not to use the therms "liquid biopsy" and "ctDNA" in an abiguous manner. See lines 158, 172/173 of the tracked changes document, where "liquid biopsy" is once used as referred to both plasma and BAL (which I find correct), but later it refers to plasma only (172/173).

Please do not use the therm "ctDNA" when referring to tissue biopsy (line 221 of the tracked changes document), as well as whenever discussed regarding BAL samples analyzed in this study.
Validity of the findings: I am mostly satisfied with Authors' comments and changes.
Additional comments: Thank you for your revision. I find this study almost ready to be published.


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

Input:
Title: EGFR MUTATION STATUS YIELD FROM BRONCHOALVEOLAR LAVAGE IN PATIENTS WITH PRIMARY PULMONARY ADENOCARCINOMA COMPARED TO A VENOUS BLOOD SAMPLE AND TISSUE BIOPSY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed most of the comments and the language has been updated.  I have made some minor edits to improve readability and will include the word document with my submission.
The addition of the demographic tables is welcome, but one glaring omission is the stage of the patients tested.  This information may help explain why the plasma sensitivity was so low. Additionally, it is currently recommended that patients with non-squamous NSCLC should undergo molecular testing, so it is unclear why the 23 patients with non-squamous histology were not included.
Experimental design: The authors have done a good job of answering my comments.
After seeing the Table 1, the proportion of patients with squamous, non-squamous and small cell seem more in keeping with what would be expected.  Also, if you include the non-squamous patients, then perhaps the proportion of EGFR positive patients would be more in line with what would be expected.  However, it is not clear why the authors did not include all patients with non-squamous NSCLC in their analysis and limited it to those with a diagnosis of adenocarcinoma as current recommendations are that all patients with non-squamous NSCLC should undergo molecular testing for EGFR.
Validity of the findings: Overall, the authors have again done a good job in answering my concerns.  However, one glaring omission remains.  The authors still insist on using the reference by Li concerning a meta-analysis of published literature.  The paper is quite old (2014) and reports on literature published between 2005 and 2013, well before the current commercial validated approved assays were available. They should find an updated reference.  Their comment on lines 190-193 that use of plasma is not a feasible alternative to tissue needs to be put in context or modified.  In advanced NSCLC, the sensitivity of plasma testing for EGFR mutations can range from 60-70% for T790M up to 85-90% for exon 19 deletions again referencing the Thress and Wu papers in the previous review.  Also, with today’s modern assays, specificity in plasma for EGFR is in the 95-99% range. There may be good reasons for the low sensitivity in this study such as Stage of disease. Earlier stage disease is less likely to be ctDNA positive in plasma than advanced metastatic disease where most of the available data comes from.  Stage is correlated with tumor burden to some degree, but as the authors have demographic data, Stage should be included.  It would be more reasonable to say that testing for EGFR mutations in BAL may be better than plasma in certain situations, like in patients with visible intra-luminal disease.  However, in advanced non-squamous NSCLC it is certainly feasible to detect EGFR mutations with acceptable sensitivity in patients who have insufficient tissue for molecular testing or cannot undergo an invasive biopsy.  The authors may want to put a line or two in speculating why the plasma positive rate was low, including the fact that this was a very limited group of patients.
Additional comments: The authors have done a good job of answering the comments and outside of the comments above feel it is acceptable for publication.


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

Input:
Title: EXPLORING THE BEHAVIORAL REACTIONS TO A MIRROR IN THE NOCTURNAL GREY MOUSE LEMUR: SEX DIFFERENCES IN AVOIDANCE
Review round: 1
Reviewer: 1

Output:
Basic reporting: In general, the manuscript is well written but I think that the first part of the introduction, as well as the description of the aims, could be improved (see specific comment below).
Experimental design: The research questions and the aims should be better addressed. The experimental design does not show serious weakness. However, some extra details in the methods section should be provided (see specific comments below).
Validity of the findings: The statistical analyses should be redone completely (see below). Thus in this state, the validity of the results cannot be evaluated. However, I appreciate that the authors recognize their study as a pilot, and they did not overstate their findings.
Additional comments: In this paper, authors tested the behaviour of the grey mouse lemur in response to a mirror image, a conspecific and a white opaque board (as control). This species is solitary and nocturnal, and the behaviour has been rarely investigated. The use of the mirror to understand the behaviour of this species can be useful in the future. The topic is interesting and the study is potentially beneficial for future researches on these (and similar) animals. However, I have a major concern and a few relatively minor comments/suggestions.
My main concern is about statistical analysis. The guidelines for analysing behavioural data strongly discourage to perform PCA. Firstly PCA should be performed on large dataset. A very general rule is to having sample size at very least three times the number of variables (see Budaev 2010, Ethology). Secondly, variance explained by two of the three PCAs are very too low. PCA analysis tend to inflate factor loadings as low and non-significant correlations can produce high PCA loadings. As expected from the variance explained and factor loadings, the PCAs with the lowest variance explained failed be repeatable. This strongly suggest that these PCAs should not be used. Instead authors choose to perform analysis by using only the data obtained from the first trial, potentially leading to data that is meaningfully interpreted. The authors can read the paper by Budaev (2020, Ethology) for an extensive review on the guidelines for using PCA. Moreover, a Dingemanse and Wright (2020, Ethology) suggest a more appropriate approach to treat variables recorded within the same trial: “A more appropriate approach would arguably be to report the correlation matrix between the different behavioural measures recorded within the same assay (within and among individuals) and then pragmatically take forward one representative behavioural measure that is normally distributed and covaries (i.e., loads) most strongly with the overall PCA component or latent variable estimated (Araya-Ajoy & Dingemanse, 2014; for other alter- native approaches, see Dochtermann & Nelson, 2014)”. Moreover, from the same paper, authors recommend to avoid using variables from the same behavioural test in PCA to infer behavioural syndrome and personality: “[..] a PCA cannot be used to imply that there is a “behavioural syndrome” or “animal personality structure” if the study does not feature repeated measures and separate behavioural assays.”. In contrast, in this study authors states that they used MIS to detect “[…] significant repeatability for the activity axis of personality” (L 51). Overall, for the above reasons, all the analyses should be redone accordingly.
Since the results could profoundly change after revising statistical analyses, my subsequent comments and suggestions will regard mainly introduction and methods.
LL 60- I think the introduction should be reorganized by providing more details on the background. Specifically, the first part of the introduction should firstly focus on the 2 general aims of using the mirror: 1) to verify whether the individuals of a certain species recognize themselves in the mirror and if not 2) to study the behavioural responses to conspecifics by using the mirror to simulate the presence of live conspecifics. The introduction would strongly benefit from clearly stating these 2 different aims.
L62 question marks is sometimes after and sometimes before brackets.
LL 71-72 please notice that some studies were also performed on highly territorial, aggressive species (e.g.  Betta splendens).
LL 78-80 I feel this is a crucial point for justifying the entire study and thus this part should be extended. For instance, how does studying the responses to a mirror image could improve our understanding of these animals? Which types of studies could be beneficiated from the use of this methods? Which information can be obtained? I agree with authors that this methodology can be useful with solitary, nocturnal species but I think this point should be addressed more extensively.
LL 97-98 the study by Cattelan and colleagues was not focused on aggressive interactions, but instead, on social interactions and sociability. There are many other studies performed in fish in which the mirror was used to study aggression: Scherer et al 2016; Arnott et al 2016; Elwood et al 2014; Balzarini et al 2014 and others.
LL 99- see my comment above on the first part of the introduction.
LL 134-135 the aim should be better addressed. I think the most important aims here are 1) to understand whether the individuals of this species recognize themselves in the mirror and if not 2) to investigate whether the mirror can be used instead of a live conspecific to measure behaviour in a more standardized and easier way. Moreover, as I have previously stated, authors cannot infer personality traits/axes by performing a single behavioural assay. Accordingly, authors should rephrase here and throughout the manuscript using “behaviour” and/or “behavioural trait” instead of “personality”/ “personality trait/axis”.
L 136 here authors should explain the term “sociability” more in depth.
LL 137-140 all these behaviours should be detailly explained. For instance, which is the biological meaning for “standing bipedally”? Aggressive posture? Social posture? For a reader not familiar with primates or mammals can be difficult why you are measuring a certain behaviour.
LL 143-144 I do not understand why you should expect that males and females behave differently. Please clearly state your hypothesis.
LL 172-173 did the two individuals were familiar or unfamiliar before the test? This is an important point as familiar individuals can behave differently from unfamiliars.
LL 174-175 please be aware that considering this test as two different tests you are violating the assumption of independence. The behaviour of an individual can affect the behaviour of the other and vice versa.
L 178 “blind” in respect to what?
LL 180-181 please state at which distance of time the two tests were performed.
LL 285 I think that the response strongly depends also on how the mirror test is performed (e.g. the presence or not of improvements).
LL 302-303 totally agree with this. Unfortunately, with this sample size it is difficult to drive any solid conclusion. And indeed I appreciate that authors did not overstate their results, but instead they recognize the limitations of the study (e.g. LL 328-330).
Figure 1: the figure would greatly benefit from showing the sizes of the apparatus.


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: EXPLORING THE BEHAVIORAL REACTIONS TO A MIRROR IN THE NOCTURNAL GREY MOUSE LEMUR: SEX DIFFERENCES IN AVOIDANCE
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this study a MIS-test was used to study personality in grey mouse lemurs. Captive mouse lemurs (N=12) were confronted either with a mirror or same sex conspecific and an opaque board as a control. Responses were summarized in three principle components representing measures of “activity”, “approach” or “anxiety”. Only the PCA representing “activity” measures was repeatable. Overall there was no sex difference in behavior across experimental conditions for PCA1 and PCA3. However, in one session males avoided the mirror and approached conspecifics more often. Hence, the authors conclude that males are more sensitive than females to the different contexts.

The Introduction covers the most important aspects of MIS-test but needs some re-structuring (see detailed comments below).  Also, the authors should elaborate a bit more why they used a MIS-test to examine personality in mouse lemurs and adapt their predictions accordingly.
L 54 Why ecological differences between sexes? They usually occur in the same environment.
L 60ff: MIS studies have originally been conducted to examine self-recognition in the context of self-awareness, e.g. Theory of mind. Therefore, it would be good to address this already in the beginning of the Introduction.
L75ff: It’s correct that nocturnal species are considered to rely more on olfactory or auditory stimuli but the MIS test is not applied to study the relative importance on the usage of visual vs olfactory or acoustic stimuli, therefore it would be good to elaborate why it would be interesting to use a MIS-test to study personality in nocturnal primates. Is the relative importance in the use of sensory modalities important to study personality?
L 78: Most MIS-tests have actually been done in captivity, hence, stating that it is difficult to study nocturnal primates in the wild is not a convincing argument.
L112-117: This has been stated already above, even with the same wording.
L134: here you should elaborate why a MIS-test is particularly suited to study personality in mouse lemurs. Why a MIS-test and not an open-field or novel object test? Explain the advantages to use a MIS-test to study personality in mouse lemurs.
L137-140: this is actually part of the Methods
L 142: explain how you measure exploration in a MIS-test.
L144: should be “… more often than females-…”
L144ff: If the intention was to study sociability with the MIS-test, you should also present your predictions accordingly. Do you assume that males are more sociable than females? If so, why? Also consider differences in social behavior between the sexes, i.e., female sleep in groups whereas males usually sleep alone.
Experimental design: The experimental design is in principle flawless. For the statistical analysis the authors should include test session as additional factor. For the PCA the authors should provide more information on the validity of the PCA (see detailed comments below).
L154: insert in groups of two to seven individuals
L 162ff: Did you conduct the experiments in the same or randomized order? If you run them in the same order include order as an additional factor to control for an order effect.
L172: Was it a familiar or unfamiliar individual?
L 190: Which criteria did you use to extract the factors of the PCA? Did you choose the PCA’s according to their Eigenvalue? If so, see Dingemannse & Wright 2020 and Morton & Altschul 2019 for some critical comments. Please also provide information on the Kaiser-Meyer-Olkin value to understand how well the three components explain the given variation.
Dingemanse, Niels J., and Jonathan Wright. "Criteria for acceptable studies of animal personality and behavioural syndromes." (2020): 865-869.
Morton, F. Blake, and Drew Altschul. "Data reduction analyses of animal behaviour: avoiding Kaiser's criterion and adopting more robust automated methods." Animal Behaviour 149 (2019): 89-95.
L184: Video-Analysis: do you have any measures for inter-observer reliability for the video analyses? Please add them.
L196: You also need to include test session as an independent factor to estimate whether order (first or second) has an influence of the response.
L197: add reference for the package nparLD
L226: Did you also check whether you find the same results when you use only the first values of PCA1 instead of the averaged values? Since you did not average the values of PCA2 and 3, it would be better to treat all values consistently throughout the analyses.
L 233: Here you present only the results of the first session but not for the second. Also, it is not clear whether the analyses for PCA1 and 3 are done only for the first or second or both sessions. As mentioned above it would be important to include session as an independent factor in your analyses.
Validity of the findings: In principle, there were no differences in behavior across the three experimental conditions, especially not between the control and the mirror and conspecific presentation, questioning the validity of the results. Especially, in light of the findings that males appear to avoid the mirror and approach conspecifics only in session one, but results of session two were not provided. So, it might be that males responded basically more strongly to novel stimuli (mirror, conspecific) than females. The results should, hence, be elaborated by adding more comprehensive statistical analyses and discussed more critically.
L252: insert “by” after determined
L 257-260: In the beginning of the sentence you state that activity as a personality trait have been found in wild and captive mouse lemurs but then you refer only to studies in captivity.
L 261: Explain what you mean that sociability is not so “easy” to quantify in a solitary forager.
L272: Here you have to explain why you did not expect to find this difference and discuss this finding regarding the social behavior of the species.
L285: Explain more specifically why it should depend on their ecology, which aspects of tehir ecology?
L287: I don’t think that you can state that nocturnal species are “non-visual”; they have a good vision to explore their environment and may only in comparison to diurnal species rely more on olfactory than visual information.
L 314: Here I would also refer to some studies of wild mouse lemurs, e.g.:
Eberle, M., Kappeler, P.M., 2004. Behav Ecol Sociobiol 57, 77–90.
Eberle, M., Kappeler, P.M., 2004. Behav Ecol Sociobiol 57, 91–100.
L319: Why should female dominance explain that females did not approach the mirror or same sex conspecifics?
Additional comments: Although, MIS-test have not yet been conducted in nocturnal primates and in principle the experimental approach is flawless, the statistical analyses have to be improved and the findings accordingly discussed.


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: EXPLORING THE BEHAVIORAL REACTIONS TO A MIRROR IN THE NOCTURNAL GREY MOUSE LEMUR: SEX DIFFERENCES IN AVOIDANCE
Review round: 1
Reviewer: 3

Output:
Basic reporting: Introduction:
-I commend the authors on including a wide variety of studies (citations) that fall outside of the primate order.
--The aim and relevance/significance of the study is not clearly stated. The manuscript would benefit from the authors clearly and succinctly conveying what the aim(s) of the study are, how their work fills a gap within this field of research, and why it is significant/relevant.
--In regards to the introduction and  background, the manuscript would benefit from a more thorough and succinct look at the relevant literature and provide more detailed examples and interpretations, so that their results/conclusions can be better and more clearly linked to former studies in the discussion. It would also be beneficial if the authors discussed some specific primate examples, other than just the one nocturnal species, Garnett’s greater bushbaby
--Line 87-95, I think this information would make more sense at the beginning of the introduction- helping to lay out what this type of research entails, why it's important, etc. Also, important to define terms at the beginning so the reader is aware and able to follow along
--In regards to animal personality, I think it would be important for the authors to include some information on how consistent personality differences can have fitness impacts on an individual
--Becker et al., 1999 is discussed and used in a comparative content in the manuscript, but the results of this work haven't been interpreted, only reported.
--Mirror tests being employed to investigate  aggression (as a personality trait) is mentioned in the introduction and the authors cite two studies (yellow-bellied marmots and guppies), but do not discuss any of the findings.
--The authors discuss how grey mouse lemurs are sexually dimorphic (females larger than males) and that females have a greater bite force than males, but do not explain what this may have to do with MIS (potential impacts), or the relevance to this study
--Lines 135-140, seems to be explaining the methods which seems unnecessary in the introduction

Discussion:
-The discussion section does not expand on/provide interpretations of their results in the context/comparison of past studies, what do the results mean (big picture). The couple of studies that are used to compare are fish studies (also need further explanation), it would be beneficial to also discuss their findings in relation to other research on primates.
--Connections to ecology are made in the discussion but it is not clear what this has to do with the results of the study
--The future work that is mentioned, again, it is not clear what the relevance and significance would be

-There are consistent grammatical errors throughout the manuscript. The manuscript would be greatly improved once addressed. Some examples where grammatical errors need to be addressed include lines 55, 81, 83, 95, 99, 101, 106, 111

-Some portions of the text are unclear and ambiguous. Thus, it is not always  obvious what the authors are trying to relay to their audience. Some examples include lines 52-54, 54-56, 112 (Are you referring to M. murinus or a different species of mouse lemur? Perhaps just use Microcebus spp., if applicable), 140 (Can the authors clarify what they mean by ‘test’ (e.g., MIS or a statistical analysis), 154, 286 (Did the authors mistakenly put ‘visual cues’ instead of olfactory?), 319-322, 333-336

-There are a few instances of spacing errors in the manuscript. Some examples include lines 62, 127, 202

-There are some inconsistencies with the in-text citations that need to be addressed. Some examples include lines 61-62, 68, 104, 108, 144

-There are a few instances where scientific names need to be italicized. Some examples include lines 369, 421, 430

-Line 75, The authors provide a citation, Becker et al., 1999  in reference to nocturnal strepsirrhines relying predominantly on olfactory and auditory cues , while diurnal species are more reliant on vision. This citation is a quite dated resource. All strepsirrhine primates use olfactory communication (diurnal, cathermeral, and nocturnal), examples from the literature below
 --Drea, Christine M. "Socioecological and phylogenetic patterns in the chemical signals of strepsirrhine primates." Animal Behaviour 97 (2014): 249-253.
--Drea, Christine M. "Design, delivery and perception of condition-dependent chemical signals in strepsirrhine primates: implications for human olfactory communication." Philosophical Transactions of the Royal Society B 375.1800 (2020): 20190264.

-Line 66 needs another parentheses after ‘fishes’

-Lines 112-114, This sentence already appears on lines 73 & 74

-Lines 114--117, This sentence already appears on lines 75-77

-Throughout the manuscript the authors use a variation (contexts, situations,  conditions, etc.) of words to refer to the three different contexts mouse lemurs are presented with (i.e., mirror, live conspecific, white board). The manuscript would benefit from consistency by using only one word throughout when referring to the three contexts.

-Lines 191-193 contains a quote, the citation does not provide a page number(s)

-Line 217 suggestion to change ‘higher’ to greater

-There are inconsistencies with how the authors refer to principal component analyses (e.g., PC1, (PC) 1). It would be beneficial to employ one way of referring to PCAs.

-There are several inconsistencies with the reference formatting. Some examples include lines 381, 393, 406, 421

-Line 331, can the authors clarify what they mean by “clearly, something different happened in males…”

-Line 131 delete ‘primates’

-The authors report some of their results in the description of Figure 2. It would be beneficial to just include the information a reader would need to interpret the figure.
Experimental design: -The study conducted by the authors fits within the scope of PeerJ

-The authors clearly lay-out their predictions for the study (lines 140-146). However, in regards to the research question, the manuscript would benefit if the authors were to more clearly and succinctly convey what the aim(s) of the study are, how their work fills a gap within this field of research, and why it is significant/relevant.
--For example, the authors state, “This pilot study opens a discussion  on ecological differences between males and females in the nocturnal solitary species and suggests males are more sensitive to the context than females” (lines 54-56). It is not totally clear to me what exactly they mean by this. There is no further discussion of what ecological differences they may be referring to.
--The only other mention in the manuscript about ecological differences appears in the discussion (lines 311-313). It is unclear to me what the authors are trying to say (i.e., the connections of ecology, ecological differences between the sexes and this study and its results).

-The authors state that this study is a pilot study (line 54) in the abstract and mention it again at the end of the discussion (line 322). However, this manuscript would benefit if a statement was included at the end of the introduction when the authors lay out what this study will entail.

-While the sample size is small (n=12), the authors recognize this and state this clearly in the manuscript. Additionally, it is not the easiest to find a large colony of captive mouse lemurs in which to perform research!

-The authors' investigation adhered to an ethical standard, which is stated clearly in the manuscript.

-I commend the authors on a detailed and informative methods section, which clearly lays out the steps necessary for future replication.
Validity of the findings: -The authors have provided all of the raw data

-All statistical analyses are robust and statistically sound. The study is scientifically sound and appropriate for PeerJ.
Additional comments: I commend the authors on a well designed study with robust statistical analyses. The figures are of high quality and labeled and described well. I especially love Figure 1! Once the language and grammar issues are addressed, as well as the aforementioned comments on the introduction and discussion, this paper will be ready for publication.

Most important points to be addressed:
 -Language and grammar
 -Aim and relevance/significance of the study
-How this work fills a gap within this field of research, and why it is significant/relevant.
-Connections to ecology to be clarified
 -Expand on/provide interpretations of their results in the context/comparison of past studies, what do the results mean (big picture).


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: EXPLORING THE BEHAVIORAL REACTIONS TO A MIRROR IN THE NOCTURNAL GREY MOUSE LEMUR: SEX DIFFERENCES IN AVOIDANCE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors improved the introduction and in general the presentation of the study.
Overall I am satisfied with the revisions they mainly did on the introduction and discussion section. The manuscript is much more improved compared to the original version.
Experimental design: After revising the manuscript, the research question and the predictions are well defined.
Moreover, the authors provided more details on the methodologies used, as requested by reviewers. Now the methodologies are fully described.
Validity of the findings: Repeatability was conducted within an interval that ranges between 10 and 22 days. This variation is extremely high and could be affected the results. My suggestion is to formally test the effect of days between the trials.

However, I am not yet fully convinced of using PCA analysis in this study, as I have extensively explained in my previous revision. The problem in using PCA in this study appears evident from the fact that only one out of three PCAs was repeatable.

The results are extensively discussed. All the points raised in the discussion sound reasonable.

Conclusions are well stated and the authors acknowledge the limitations of the study and avoid overstating the results.
Additional comments: L191 add "sexually" before receptive

L 387 remove one "s" in contexts

L 458-459 these scores are in line with those found in other species (in general repeatability for behaviors is quite low).

L 576 correct "sensitive"

Figure 1 I would suggest adding sizes (in for example cm) to the figure.

Figure 2 Adding asterisks to the figure would improve readability.


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: EXPLORING THE BEHAVIORAL REACTIONS TO A MIRROR IN THE NOCTURNAL GREY MOUSE LEMUR: SEX DIFFERENCES IN AVOIDANCE
Review round: 2
Reviewer: 2

Output:
Basic reporting: Abstract:
-	There are some grammatical and spacing issues that need to be addressed. For example, Lines: 19, 21, 22, 24
-	Lines 23-24, the wording is a bit unclear- ‘…whether mirror presentation elicits a similar reaction than a conspecific…’
-	Suggestion to mention that this work is a pilot study earlier in the abstract, perhaps Line 21
-	It is still unclear to me how this study/results relates to ecological differences in male and female grey mouse lemurs (Lines 29-31)

Introduction:
-	There are some grammatical and spacing issues throughout the introduction that need to be addressed. For example, Lines 34, 37, 42, 46, 48, 76, 79
-	Suggestion, Line 35 replace ‘able’ with capable
-	Suggestion, delete ‘already’
-	Lines 41-44, wording is unclear and confusing, suggestion to delete ‘capable’
-	Line 42, can the authors provide a couple examples of ‘social’ and aggressive behaviors’ (e.g., XXXX)
-	Line 42, consider adding the species’ names (primates, cetaceans, etc) after ‘only a handful of species…’
-	Can the authors clarify what they mean by ‘visual attractiveness’? (Line 48)
-	Consider making two sentences instead of one, Lines 48-51
-	Suggestion, delete ‘Interestingly’, Line 67
-	Lines 77-80, can the authors make a statement of interpretation about the results of the bush baby study. For example, what is or why is it significant that bush babies showed threat and bipedal postures when close to a mirror?
-	Lines 80-83, This sentence is unclear and confusing. Why do nocturnal strepsirrhines, which are more reliant on olfactory and auditory cues, ‘appear sensitive to mirror-image simulations’?
-	Suggestion, Line 85, delete the first ‘the’ & ‘forest’ and ‘coasts’ should be plural
-	Line 93, fix phrasing ‘show threat’
-	Line 100, ‘seems’ should be singular
-	Line 107-108, can the authors clarify and explain what they mean by ‘…origins of known differences between the sexes’
-	Suggestion, Line 109, Change to ‘In this study we aimed to…’
-	Suggestion, Line113, Change to ‘We predicted that’, delete ‘that’ after (1)
-	Suggestion, Line 113, Change to ‘this experiment will allow for the detection of repeatable reactions across time and space’. Delete period at end of sentence and replace with a comma
-	Suggestion, Line 114, delete ‘we also predicted that’, change to ‘individuals will present a similar reaction toward a conspecific and the mirror’. Delete period at the end of the sentence and replace with a comma
-	Suggestion, Line 116, delete ‘Finally, we predict’. Change to ‘males and females will differ their responses in the mirror context (Mir).’
-	Suggestion, Line 117-118, Delete sentence beginning with ‘Specifically’
-	Suggestion, the authors may want to consider mentioning that this study is a pilot study earlier in the introduction, instead of the last sentence

Materials & Methods:
-	Suggestion, Line 127, consider changing ‘The room’ to ‘Aviary temperatures were maintained…’ use aviary for consistency instead of room
-	Suggestion, Move sentence ‘We tested twelve…’, Lines 129-133 to appear under the Behavioral Tests subheading, perhaps at the beginning of Line 141
-	Suggestion, Line 133, consider rephrasing ‘Animals were individually identified via ear tags’.

Behavioral Tests:
-Line 145, add ‘is’, so it reads ‘is visible’
-Line 146, should read ‘age and sex class’
-Line 148, consider changing cage to aviary for consistency

Statistical Analyses:
-	Add (PCA) after principal component analysis
-	Line 164, consider rephrasing to read ‘The first three…’
-	Line 165, delete comma before ‘accounted’
-	Line 166, suggestion to add comma after ‘variance’
-	Line 169, add comma after ‘Subsequently’
-	Line 171, wording is a bit unclear
-	Suggestion, delete ‘Then’

Results:
-Line 199, Can the authors clarify what they are referring to when they state, ‘estimator of this behavior’? Are they referring to ‘activity’ as mentioned in Line 194?
-Line 213, add comma after ‘together’
-Line 232-234, The phrasing here is a bit informal. The authors might consider rephrasing this sentence.
-Line 232, Can the authors explain what the possibility of relating their ‘activity score’ to the activity axis of personality would mean or indicate in this context?
-Line 240, delete ‘the’ and ‘of’ and change ‘approach’ to ‘approaching’
-  Line 242, delete ‘for example’
- Suggestion, consider citing the paper in-text, Line 256-257
- Line 280, add comma after  (2019)
- Line 281, ‘Theory of Mind’ does not need to be capitalized
- Line 285, add comma after ‘In this species’
- Line 285-287, consider rephrasing as this sentence is unclear/confusing
- Line 287, add comma after ‘species’
- Line 288, consider deleting ‘Interestingly’

Discussion:
- Lines 219-227, This paragraph seems like a repeat of the results and not an interpretation of the results reported.
- Line 297, Can the authors clarify what they mean by ‘wild animals’? Are they referring to wild mouse lemurs?
- Line 290-293, The wording here is confusing, consider revising and explain and/or give an example of what is meant by ‘social function’
-  Line 300, ‘group’ should be plural
-  Line 305-307, This sentence seems out of place
- Line 309-310, consider rephrasing ‘As a follow up to our pilot study…’ and consider revising the wording, as it seems informal. Suggestion to delete ‘interesting to’
- Line 314-316, The wording here is confusing and not clear, consider revising
- Line 317-318, change to ‘between the sexes’
- Line 318, test should be plural

Conclusions:
-	Lines 328-340, Language/wording needs rephrasing.
o	It is not clear as to what the significance is in relation to the study. The authors state that this pilot study aimed to ‘describe the response of M. murinus toward a mirror presentation and to explore whether they responded as when in the presence of a conspecific”. Why is this important? What is the significance in understanding this? What gap in the field/literature does this fill and/or contribute to?
o	How would hormone analyses contribute to the documented reactions of the study subjects? How would they ‘explain sexual dimorphism in the reaction toward the mirror? Same as for PET scans, how does this improve/advance our knowledge?

References:
-	Formatting and spacing issues throughout the references. For example, Lines 348, 368, 377, 386, 396

Legends:
-	Spacing issues, Line 539
-	Line 541, change ‘arena’ to area
-	Spacing issue, Line 546
-	Line 547, add ‘and’ after ‘panel;’
-	Can the authors clarify what they mean by ‘situation’, Line 549. Does this refer to context?
Experimental design: The study fits within the scope of Peer J.  The authors’ investigation adhered to an ethical standard, which is stated clearly in the manuscript.
Validity of the findings: The authors have provided all of the raw data and state that their script will be provided freely upon request (see reply to reviewers). All statistical analyses are robust and statistically sound.
Additional comments: I commend the authors on their work addressing reviewer suggestions. However, there are still a number of issues with the current manuscript which need to be addressed.

Most important points to be addressed:
1.	Consistent language/wording/phrasing issues throughout the manuscript
2.	Grammatical and spacing issues throughout the manuscript
3.	Aim/purpose of study is still not clear
4.	Discussion- restates the results, there needs to be more interpretation of the results and comparisons of them to other relevant work (to put this work in context)
5.	Conclusions- overall conclusions and their significance of the present study is not clearly stated. It is not clear how the ‘future work’ that is suggested would improve/advance knowledge.


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript by Bu and Huang et al. entitled "Identification of Vitamin D-related gene signature to predict colorectal cancer prognosis" constructs a vitamin D-related gene signature to predict prognosis in Colorectal cancer (CRC), which is one of the most common malignant carcinomas worldwide with poor prognosis. Vitamin D and vitamin D-related genes play a vital role in CRC.
Experimental design: Very well wrote.
Validity of the findings: TCGA data validate with GEO set
Additional comments: The authors wrote this manuscript very well.

Need to highlights D-related signature gene in discussion and wrote previous studies related to those in CRC or other cancers in detail.


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The concerns are in the attachment.


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript ' Identification of Vitamin D-related gene signature to predict colorectal cancer prognosis’ by Bu et al using a Vitamin D related gene signature model to predict the prognosis in colorectal cancer. However, there are some concerns which the authors need to address in this manuscript.
The authors need to improve their language used in this manuscript, as it is very simple in many places rather than being scientific.
Experimental design: The authors have clearly indicated that they have obtained the clinical information of CRC patients. There is no proper information regarding the source and there is no statement on whether this study was reviewed by an institutional ethics committee or board.
Validity of the findings: The authors should describe more in detail about the univariate cox analysis performed in the CRC samples and present those results. It is not clear in the results how those 17 genes were selected. Also, it is not clear as to how and on what basis those 17 genes were classified further as high and low risk genes.

The construction and validation of this gene analysis has been based on the ‘hazard ratio’, but it has not been made clear anywhere in this manuscript as to what this means and how this is calculated. This is very important because this is the foundation for this model.

Under the high risk and low risk genes classification, the authors have 14 and 3 genes respectively. The authors should explain the biological importance of both these genes and provide more evidence as to why they are important.

The authors description of the univariate and multivariate analysis in table 3 is not clear as stated in the text of the manuscript. Also, it is not clear as to what motivated the authors to perform this analysis.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors fulfilled the comments and I do not have further concern.  The revised manuscript looks great and acceptable, after other reviewer's comments.
Experimental design: The authors provided details in a revised version of the article.
Validity of the findings: They also validate their data in more than one database.
Additional comments: Thanks for providing responses.


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The quality of this article has improved considerably post-revision. Figures also have a better resolution.
Experimental design: The authors have corrected concerns about experimental design and information provided.
Validity of the findings: The results have now been validated by adding more datasets thus making the conclusions more reliable and less-extrapolated.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF VITAMIN D-RELATED GENE SIGNATURE TO PREDICT COLORECTAL CANCER PROGNOSIS
Review round: 2
Reviewer: 3

Output:
Basic reporting: The manuscript “Identification of Vitamin D-related gene signature to predict colorectal cancer prognosis” by Bu et al using a Vitamin D-related gene signature model to predict the prognosis in colorectal cancer. However, there are some minor concerns which the authors still need to address in the revised manuscript.
Experimental design: N/A
Validity of the findings: 1. Appreciate the authors on providing the formula for the hazard ratio. Can the authors validate this formula as to how or on what basis these formula was formulated?

2. Under the high risk and low risk genes classification, the authors have 14 and 3 genes respectively. The authors should explain the biological importance of both these set of genes and provide more evidence as to why they are important.
The response provided by authors for this comment is not satisficatory.
The main reason to ask this question was to understand the context of these genes in terms of their functions in other diseases, so that, it will aide in the analysis in the context of CRC. However, the authors fail to discuss this point clearly again.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF THE BIOLOGICAL FUNCTION OF MIR-9 IN SPINAL CORD ISCHEMIA-REPERFUSION INJURY IN RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This work reports on the bioinformatics analysis and experimental validation in identifying the biological importance of miRNA miR-9 for the treatment of neurological disease Spinal cord ischemia-reperfusion injury, multiple cases of which have been already reported in humans. The study was carried out in the Rat model. The study revealed the association of miR-9 mimic with multiple signaling pathways of Spinal cord ischemia-reperfusion injury and can be a potential therapeutic agent for it. Gene Ontology and KEGG bioinformatics analyses were also performed to identify potential target genes of miR-9 that are associated with important biological processes and signaling pathways. Considering the recent developments in miRNA-based therapeutics, this present study can be considered as an important contribution to the field. This work is within the scope of the journal. The authors used the English language professionally throughout the manuscript and the manuscript was appropriately structured following PeerJ standard. Figures are of publication quality and appropriately annotated. Raw data is available as supporting information files.

To aid the readers, a little more background on the Spinal cord ischemia-reperfusion injury disease should be added. Please discuss a couple of case studies on Spinal cord ischemia-reperfusion injury (For instance: https://dx.doi.org/10.7759%2Fcureus.5279).
Experimental design: The objective of the study is well defined and experiments are well structured and seem to be performed accurately following standard strategies.

A little more background details on GO and KEGG analysis approaches should be provided in the “Materials and Methods” section, and the reasons to choose these approaches (compared to other similar approaches) should be briefly described.
Validity of the findings: The results are presented stepwise in a logical manner and also in detail. Statistical analyses have been performed and reported. The results are well discussed answering the original research question.

A separate “Conclusions” section should be added to summarize the results and any future application of this workflow and applicability of this work should be discussed.
Additional comments: No additional comments


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

Input:
Title: IDENTIFICATION OF THE BIOLOGICAL FUNCTION OF MIR-9 IN SPINAL CORD ISCHEMIA-REPERFUSION INJURY IN RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Basic reporting
1. I thank the authors for their efforts to write the manuscript in clear and professional English. However, the manuscript could use grammar and language check. Grammatical errors are seen in multiple places. Some examples are lines 34-37, 211-213, 242-244, 260-261.
2. I appreciate the authors’ efforts to set the context for the study in the introduction section. The manuscript could benefit from further elaboration about why the authors chose to study mir-9 compared to other microRNAs like mir21 that might be involved in SCII and other neurodegenerative diseases. Much of this context and background is provided in the discussion section (lines 229-240). A better place for this background would be the introduction.
3. Although the authors reference multiple papers related to mir9’s role in neuronal injury and disease, they leave out a previous paper on mir9 that investigates its role in ALS- Zhou et al, 2013 have shown that mir9 has a important role in the pathogenesis of an ALS model. Interestingly that paper showed that mir9 is upregulated in the ALS model as opposed to the downregulation seen in SCII that the authors report in the current manuscript. The authors could discuss and shed some light (even if just speculation) as to what these differences could mean in terms of the biological function of mir9 in these situations.
4. Font sizes within figures need to be larger for clarity. For examples in figure 3A, the font is very small and difficult to read. Figure and table legends need to be more descriptive. Example : Table 1 has no legend description other than a short title for each section.
5. I commend the authors’ efforts to summarize MAPK and Notch2 signaling pathways in figure 4. However, it is not original data or model of a mechanism being uncovered in this paper. It appears to be   a summary of current knowledge of MAPK and Notch signaling pathways. This would be unnecessary and can simply be referenced.
Experimental design: 6. The authors report that mir9 expression is downregulated 48h hours after SCII, but don’t discuss the 72 hour timepoint at which mir9 expression almost goes back up to normal compared to sham group. Is this time point significantly different from sham? It would be beneficial to know what happens after the 72 hours time point. Does the expression of mir9 go up and stay high after 72 hr? If that is the case, the downregulation of mir9 seen after SCII is pretty short lived. What would this mean for the pathology of SCII and the benefit of therapeutic intervention by targeting mir9?
7. Major comment - While this manuscript uncovers a possible mechanism of action of mir9 in spinal cord ischemia-reperfusion injury (SCII), it does not investigate this mechanism in a rigorous and compelling manner. The authors find the possibility that mir9 might be acting through a MAPK (MAP2K3) pathway or Notch pathway to confer reduced BSCB disruption, inhibit apoptosis and show reduced inflammatory response after SCII, but do not clearly show that mir9 acts through either of these pathways to exert these effects.
A compelling experiment that would test this possibility is to determine if a mir9 inhibitor shows the opposite effect to mir9 mimic on neuroinflammation and apoptosis, and these effects are rescued by a  MAPK inhibitor. For examples, Yao et al showed that mir21 has a neuroprotective role after SCII via a MAPK signaling pathway by targeted inhibition of MAP2K3. In this paper they did a series of experiments utilizing mir21 mimics, inhibitors and MAP2K3 inhibitor SB203580. The authors of the current paper could utilize this or a similar MAPK inhibitor to definitively test if mir9 acts through MAP2K3. Additionally, Yao et al showed that mir21 interacts with the 3’UTR of MAP2K3 using a luciferase reporter assay. The current manuscript could use an additional experiment to validate the authors’ hypothesis that mir9 acts by downregulation of MAP2K3.
Similarly, they could show the involvement of Notch2 in this pathway by inhibition of Notch2 using siRNA as done by Meng et al, 2015.
8. Acronymns/abbreviations are not expanded the first time they are used. For example BSCB, KEGG. GSE138966 – What is it? This is not explained clearly to a non-expert reader the very first time it is mentioned in the manuscript in lines 102-104
9. Lines 171-73 could use further explanation about the rationale for conducting bioinformatic analysis.
10. Supplemental table 1 gives a list of 327 potential target genes of mir9 apparently from two different databases. It would be helpful to include additional information in this table other than just the gene name such as – the database that it came from, the selection criteria used by the database, scoring if any, description of the gene name, etc
Validity of the findings: 11. The data provided are robust and clearly presented. I commend the authors for their work and effort to present the complete raw and analyzed data.
12. A separate conclusion section is not provided. There is a short summary at the end of the discussion section. The authors do not overstate the findings. However, the authors draw the conclusion that mir9 may be a potential target for reducing neuroinflammation and apoptosis after SCII. The results certainly present a possibility for it but it remains to be determined whether mir9 really acts through the proposed pathways. It could be a little premature to state that mir9 could be a potential target for therapy before a mechanism of action is established.
Additional comments: This manuscript presents an interesting possibility for a mechanism by which mir9 affects neuronal outcomes after SCII. While the data in this manuscript suggests a mechanism of action for further investigation, it does not establish this mechanism in a rigorous and compelling manner. The authors could have done a few more experiments to take it to the finish line. However, the data presented are sound and the authors have done their part in presenting all of the data. On its own, this manuscript is acceptable with revisions as suggested below.


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: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The subject of the manuscript is interesting and the experiments are well performed. The results are believable. However, the manuscript is, at times, wordy, especially the discussion that must be shortened.
Experimental design: Correct
Validity of the findings: OK
Additional comments: The subject of the manuscript is interesting and the experiments are well performed. The results are believable. However, the manuscript is, at times, wordy, especially the discussion that must be shortened. The Introduction needs more clarifications for the benefit of a general reader. I suggest to substantially modify the manuscript in the sense that it needs a better Introduction and a much shorter Discussion, or even a combination of Results and Discussion. The paragraphs between lines 256 and 284 could be deleted, and also the last phrases from lines 287 (“Investigating…) to the end of the Discussion.
Minor points:
Line 51: Why is a ‘minimal’ operon? Is there any maximal one?
Line 53: Explain what UV irradiation does to the DNA
Line 65: It should be stated from the beginning that the operon is encoded by a plasmid, so that it can be horizontally transferred.
Lines 204-205: Did the survivals resume normal growth after irradiation?
I found some small typos in lines 71, 229, and 245.


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: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The structure of manuscript by Janosikova et al fits in to PeerJ requirements. The introduction demonstrate enough background and literature is relevant. English is plain and acceptable, with some misspellings like on line 243, some inconsistency throughout the text like UVC and UV-C and missed intervals between words like in legend for Fig. 2. Figures are relevant to the content of the article, but not all of them have a good quality. There is not enough explanations for Tables 4 and 5 and it is not clear if they are fully presented because there are no right margins for both of them. Raw data Excel file named “Percentual survival after 120 s (300 mJ/cm2) exposure of UV-C” is provided, but presentation of data in that file is the same as in table 3. Is there any reason to show in a separate file the same numbers as in Table 3? Why Excel was chosen for presenting those eight numbers? There are no calculations or any sorting…
Experimental design: Authors understand (lines 291-292) that standards in the field of radiation resistance is presenting radiation dose-survival curves. Such experiment requires just three days of work and it will validate authors’ conclusions extensively.
There are some major confusions in the description of methods.
1)	In the abstract authors report that the large fraction E. coli survives 120-s exposure to UVC light and provide number for radiant exposure 300 mJ/cm2. It sounds like too high radiant exposure for about 20% E. coli survival (please compare with Anita Krisko and Miroslav Radman “Protein damage and death by radiation in Escherichia coli and Deinococcus radiodurans” www.pnas.org/cgi/doi/10.1073/pnas.1009312107 .
Indeed, from Materials and Methods section we can see that radiant exposure was estimated at the UVC device aperture, not at 60 cm distance from UVC lamp where samples were irradiated. So, to avoid the confusion, authors should not use the number 300 mJ/cm2 without explanation about 60 cm distance. Of course, it would be better to measure radiant flux directly where samples were by using UVC light dosimeter.
2)	It is unclear how the authors described their calculation of CFU/ml. They irradiated 5 ml of cell culture in a small Petri dish. After UVC radiation they plated 0.01 ml of those cells. It means that CFU/ml = number of colonies at certain dilution x dilution factor x 100. It is not clear why they divide on volume of culture plate (mL).
3)	It is not clear where a legend is for Supplemental Figure 1. Some explanations for Suppl. Fig. 1 are needed: why DNA of pLK18 plasmid is almost invisible and why two DNA MW markers are shown?
Validity of the findings: The manuscript adds to the understanding of the role of minimal ter operon to radiation resistance of bacteria. Authors hypothesize that minimal ter operon is important for E. coli UVC resistance. In general, the data presented in the manuscript look novel and interesting. The results are relevant to the hypothesis and support authors’ conclusions. However, their presentation must be improved significantly prior to publication. Importantly, the additional experiments are needed. The authors should provide survival curves for both strains irradiated by UVC. Very important that the authors should use radiant exposure numbers more accurately.
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: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: The paper presented results on the UV-survival and DNA mutation rate of E.coli cells complemented with ter operon, genes that proved to give resistance to tellurium and other stress.
Some parts need clarification, but in general, the manuscript is well written.
Nevertheless, the results are little described and poorly exploited. Tables and figures are of bad quality or not appropriated to present the results.
In the discussion, the authors confront previous work and evidence related to the ter operon as a summary or review of prior work. Still, in the end, there is no clear explanation of why the cells that carry the ter operon are more fitted under UV than the ones lacking these systems; maybe complementary assays will be needed to arrive at a more advanced conclusion.

The raw data of sequences are shared in the database.

The literature references are sufficient and indicated the lack of reports and the importance of studying the relationship between UV resistance and ter operon.
Experimental design: The original primary research fits within the Aims and Scope of the journal.
The experimental procedures are relatively simple, UV-exposure, plating, UFC counting, with the addition of detecting mutations by WGS sequencing different exposed colonies with and without the operon.
The methods are described with sufficient detail & information to replicate.
The research question: are ter genes improving/related to UV-resistance in E.coli? is it only answered at a basic level. The authors showed that there is evident more growth after UV when the cells have the ter operon and that this correlates with more mutations in their DNA. But they do not perform other experiments that can help provides clues on why is that: oxidative stress markers, ultrastructural changes, overexpression of repair genes, DNA photoproducts measurement, etc.
Validity of the findings: As I said before, the authors explored the UV resistance due to the occurrence/expression of ter operon in the transformed cells and showed evidence on these regards. But it is not clear the mechanism (lack of further experiments), but there is no clear explanation of this behavior with contrasting data from previous works.
The results are poorly exploited, and the figures/tables are not informative enough.
The manuscript is too repetitive in all its sections.
Additional comments: The authors will need to exploit the results they already have and learn to present them more attractively, especially on the results of the WGS of each genome.
Figure 1 is ok.
Figure 2 is incomprehensible.
Tables 1, 2, and 3 are unnecessary as those results are covered in Fig 1.
Tables 4 and 5 should be combined, improved for clarity, and better explained in the text.


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: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: I found that the authors have addressed properly all the concerns raised by the reviewers, and I find that, in its present form, the manuscript is acceptable.
Experimental design: OK
Validity of the findings: OK
Additional comments: I find that my concerns have been properly addressed.


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

Input:
Title: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript by Janosikova et al fits in to PeerJ requirements.
Experimental design: Authors corrected the mistake in the description of their of CFU/ml.
Accepted.
Validity of the findings: The manuscript by Janosikova adds to the understanding of the role of minimal ter operon to radiation resistance of bacteria. Authors hypothesize that minimal ter operon is important for E. coli UVC resistance. In general, the data presented in the manuscript look novel and interesting. The results are relevant to the hypothesis and support authors’ conclusions.

In my first review I suggested to perform additional experiments building survival curves for both strains irradiated by UVC.
The authors replied that it is not possible in their lab: "If we assumed that cell viability would decrease with increasing radiation (since only 4% of Ter- negative cells and 20% of Ter- positive cells survived after 2 minutes of irradiation), then we would have to reduce the radiation dose. We do not have another UVC emitter available in our laboratory, which would allow us to set the desired doses needed to create a credible survival curve."
The explanation sounds strange to me. I do not see any problem in UVC radiation for periods of time shorter than 2 min. It is easy to turn off UVC light in biological safety cabinet after seconds of UVC exposure.
However, I understand that it is not easy to plan experimental work in the present time of COVID-19 and accept authors refusal.
Additional comments: When situation allows, please consider building full survival curves in your experimental work like presented in this 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: RESPONSE OF ESCHERICHIA COLI MINIMAL TER OPERON TO UVC AND AUTO-AGGREGATION: PILOT STUDY
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors have improved the manuscript, on respect to the structure and general discussion. But now, the manuscript results are only one table and one growth figure.
As the authors say in the text the work is a pilot study, for me this is too preliminary and incomplete to be published at this stage.
Experimental design: The authors did not perform any new experiment suggested by the reviewers
Validity of the findings: The work is incomplete as it is presented.
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: PSYCHOMETRIC PROPERTIES OF THE SWEDISH VERSION OF THE SATISFACTION WITH LIFE SCALE IN A SAMPLE OF INDIVIDUALS WITH MENTAL ILLNESS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an important and interesting study and I appreciate how you choose to focus on the salutogenesis and not the pathogenesis when working on modern management of mental illnesses. You use a clear professional English language and literature references. You background puts the paper in context.
Experimental design: Please describe why you have chosen to use the SWLS instead of many other rating scales for well-being (LiSat for example). On line 83-84 you present SWLS as “for instance” which gives me the feeling that it was a coincidence that you chose this particular rating scale. Could you argue shortly why this scale should be used, for example as one of the most commonly used or that it is easily available to use in clinical situations. Something that tells me why this of all scales should be evaluated.
I think you have argued and described the difference between CTT and IRT, but I miss a discussion about test-retest evaluation. You have chosen a population with mental illness such as depression, bipolar disease or personality disorders. Test-retest reliability would be very interesting in this population since well-being can be affected by mood and affections which can be of great variation in some mental- illnesses. Is it stable to use at any time or could it be different result using it a week later?
How did the survey take place? A postal survey or at the out patient clinic? Did they fill it in by themselves or with help of others?
What were the different diagnosis in the population? Mental-illness is a very wide concept and a more specific details would be preferable. Where there affective disorders, psychosis, personality disorder. I would like to see a bit more detailed description of the population. For example no one would accept a description of “neurological disease” when evaluation a rating scale, it would be needed to specify for example Multiple sclerosis or chronic neurological degenerative disorders. Mental illness is very wide and it does not make the patients right to not discriminate this better. I also think that the scale could differ in different mental disorders so I would like to see a more described population.
Validity of the findings: There are a lot of numbers and percentages in the participants-section and the reading does not enables of these numbers but instead are a bit difficult. Could the population be described in a table?

The findings are well presented and argued for. What I miss is an very important part of psychometric properties, the data completeness.   if there are a lot if missing data it suggests that the participant does not understand the questionnaire or are not able to use it properly. I miss comments on missing data, I doubt that over 200 individuals actually filled in every item adequately, there usually are some missing data.

As mentioned before I also miss test-retest reliability in this particular population, or have the population described better. You can also describe and argue why no test-rest was made and that could be sufficient, but looking from a clinical perspective I argue that both data completeness and test-rest evaluation are more important than some of your result. If the result differs from one week to another, is it then life satisfaction that are measured or something else? and if a lot of data are missing can you then rely on the scale? This could be discussed in the limitation section or for future research.
Additional comments: Taken together, it is an important study but I miss some details in the methodology and description and presentation of the population. I think that by only name the population “ a sample of individuals with mental illness” you minimizes the patient group as packing them together in a very broad concept. I suggest a revision as described above and I think this could make the article more appealing for the reader and it could strengthen its credibility.


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: PSYCHOMETRIC PROPERTIES OF THE SWEDISH VERSION OF THE SATISFACTION WITH LIFE SCALE IN A SAMPLE OF INDIVIDUALS WITH MENTAL ILLNESS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors present an English well-written study, with a robust analysis of the psychometric properties of the Swedish-adapted version of the life satisfaction scale in a sample of individuals with mental illness.
The methods are adequate and consistent with the objective of the research that has been clearly stated in the introduction.
The tables and figures presented provided detailed information about the psychometric study.
Experimental design: The research question is clearly defined and aligned with the aims of the journal.
This study presents a robust methodology, and a well-performed statistical analyses, that constitutes a strength of the manuscript from my point of view.
Validity of the findings: The authors have constructed an introduction to put the objective of this research into the background. It is clearly identified that this instrument has not been used previously for the purposes that are intended to be explored: measuring the satisfaction with life in a population with mental illness, with or without occupation.
Conclusions are well supported by the analyses and are well connected with the research question.
This will allow mental health professionals in Sweden to use this tool in a standardized way with their patients. It would be interesting to continue researching to provide thresholds of interpretation or classification that improves their use in the clinic, such as minimal clinically important change, ...
Additional comments: The authors present a well-written study, with a robust analysis of the psychometric properties of the Swedish-adapted version of the life satisfaction scale in a sample of individuals with mental illness.
I would like to congratulate the authors on the work. The factor validity analyses, especially in the confirmatory and invariance study that are presented, are robust and methodologically well-executed, which generate interesting conclusions for the practical application of this scale in this study population. The tables and figures presented provided detailed information about the psychometric study.
Please, find below some comments and questions about the manuscript that have arisen after careful reading, and whose objective is to contribute to the improvement of the manuscript readability:

•	In the method section lines 165-169… you have used the “listwise method to have the missing data”. Could you provide readers more information about this process?
- Could you provide information about the sample size used to perform this analysis?. Is there a methodological justification? Is the sample size sufficient to support the analyzes carried out? Thus, describe it in the text to give more power to the results.
•	Line 178. The value of Cronbach’s Alpha could perhaps be placed in the results section, rather than in the method.
•	Line 180, Statistical Procedure. When you apply exploratory factor analysis using principal component analysis with SPSS, could you indicate if you have used any type of rotation (Varimax, etc.) in the process?
•	Line 199, Results-Exploratory factor analysis. Can you indicate in the text the percentage of the variance explained by the one-dimensional solution?
•	When you write that only one factor with eigenvalues greater than 1 was found, perhaps they should explain this factor extraction condition previously in Methods.
•	Line 222 and 232. Use of abbreviations. Define CFIs, before using it in the text for the first time.
•	Line 237-238. When you describe the discrimination values (from 1.95 to 3.81), could you indicate which specific parameter/index you are referring to?

Thank you very much,
Sincerely


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: PSYCHOMETRIC PROPERTIES OF THE SWEDISH VERSION OF THE SATISFACTION WITH LIFE SCALE IN A SAMPLE OF INDIVIDUALS WITH MENTAL ILLNESS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The text is generally well structured and contains valuable information. However, I have some concerns of minor details, and remarks on references. However, I also have one major reflection concerning the concept of life satisfaction in relation to the dichotomous aspects of pathology and health. The authors could be clearer in the introduction section in defining what the instrument SWLS is measuring. This can clarify the concept life satisfaction in relation to subject well-being and quality of life.

My considerations:
Abstract:
•	Line 29: The reference to WHO (2001) is accurate in relation to mental health, but the original reference should be WHO (1948). The WHO (2001) reference is also given in the introduction, and as this reference concerns a shift from focusing disease to illness, or a medical to salutogenic approach, it seems vital to the current context in measuring life satisfaction. In Ottawa Charter (1986) this aspect of health is further developed and could be considered for reference inclusion in the introduction section.
•	Line 39-40: I find it unnecessary to include the measurement information for SWLS in the abstract section.

Introduction:
I suggest that the text should be scrutinized and changed in some sentences. In the introduction section, e.g. (Line 68) “In Sweden, approximately 50% of people who are on sick leave are on sick leave due to mental illness.” Suggestion: About half of the proportion on sick leave in Sweden are due to mental illness. (Line 108) “….weigh more or less in relation to …”. Suggestion: …. might be of varying weigh in relation to …”.
(Line 77): Consider alternative or complement to the reference WHO (2001), also given in abstract. The WHO (2001) refer to WHO (1946) (From reference WHO (2001), page 3:“The importance of mental health has been recognized by WHO since its origin, and is reflected by the definition of health in the WHO Constitution as….).

Some considerations regarding references:

Article or chapter by Pavot (2018), and articles by Jovanovic et al (2020), and Pires et al (2013), are missing in the reference list.
The articles Nima et al (2020 a and b) are presented in the text with different publication years (2020ab and 2021), or is there a missing reference?
In the text references for Lee (2014), and Sachs (2004), are presented in the reference list with other publication years (Lee (2018), and Sachs (2003)).
There are different publication years for Kline, R. B. (2005 in the text, and 2018 in reference list (different editions?)). The same difference is noticed regarding Tabachnick and Fidell (in the text 1996, and in the list 2012).
Experimental design: The design and statistical methods is clearly described and suitable for the study. Some minor remarks:
Line 163: I suggest that the foot note is implemented in running text.
Line 165: Replacing the term “drugs disorder” with substance use disorder.
Validity of the findings: I have no concerns regarding the well-structured and explained statistics and results. The tables and figures give complementary information to the text. Raw data has been reviewed and found accurate.
Conclusions and suggestions for further research seems well established.
Additional comments: Thank you for the commendable analyse of this highly used instrument in quality of life research. The results will be, as you have pointed out, of great value in the future research.
I have just a few remarks in my review but ask you to correct the references


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: PSYCHOMETRIC PROPERTIES OF THE SWEDISH VERSION OF THE SATISFACTION WITH LIFE SCALE IN A SAMPLE OF INDIVIDUALS WITH MENTAL ILLNESS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I have no further questions or comments.
Experimental design: I have no further questions or comments.
Validity of the findings: I have no further questions or comments.
Additional comments: The authors have responded very well to my comments and have added supplementing details in the article adequately. The inquires that could not be fulfilled are well argued for and/or commented as a limitation. I think this is a well-written and interesting article on an important topic. After the revision it has a better credebility and I think it is ready to be published.