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

Input:
Title: THE EXTENT TO WHICH CANCER PATIENTS TRUST IN CANCER-RELATED ONLINE INFORMATION: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: •	This manuscript should be reviewed by a native English speaker to review multiple errors (grammar and tenses). The authors should also proof-read the manuscript to correct errors e.g. Line 39: “therefore, physicians and other health care providers have to should…”, Line 370: “…between females and males was quiet small”

•	The introduction supports the rationale for the study. However, in line 100, the authors state that credibility is only one trust component but do not elaborate on their stance (definition or description) on trust

•	The results are relevant to the hypothesis. The authors rightly point out that the available evidence does not support conclusions on the second part of their hypothesis.
Experimental design: •	The research meets the standards for conducting systematic reviews. However, the authors’ method for rating the quality of studies is questionable. The RTI item bank used requires studies to clearly state their inclusion criteria. However, as stated in Line 267, four of the included studies did not provide details on their inclusion criteria. The authors should clarify what they hoped to achieve by using the RTI criteria to assess the studies. They should also provide reasoning for choosing some of the criteria and not others.

•	In line 152 the authors state 'participants' trust' and later 'active trust', how do they distinguish between the two?

•	In line 156, the full text screening was divided into 'study characteristics' and 'report characteristics'. Yet, the authors do not specify which criteria falls into which group in the subsequent text.

•	In line 135, the authors should define what PICOS stands for and their reasoning for choosing certain aspects of the acronym and not others

•	There is some redundancy between the information in 'Eligibility criteria' (line 146) and 'data extraction and quality assessment (line 166). E.g. both line 154 and 179 states that two researchers reviewed the studies

•	in line 111, the sentence isn't clear and may be misconstrued
Validity of the findings: •	The hypothesis of this study is relevant given that it approaches the topic from a systematic review perspective. However, since the quality of the studies included in the review is somewhat questionable, it dilutes the value of conducting a systematic review.

•	The conclusion rightly stated that there is little evidence to answer the second question they sought to answer. However, going by Table 2, it appears the other conclusions may be overstated. Rather than attempting to make broad conclusions, the authors should be more subtle about the generalisability of their findings
Additional comments: No additional comments


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

Input:
Title: THE EXTENT TO WHICH CANCER PATIENTS TRUST IN CANCER-RELATED ONLINE INFORMATION: A SYSTEMATIC REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: Good.
Experimental design: Good.
Validity of the findings: Good.
Additional comments: While the Introduction is well-written and covers important research pertaining to cancer information seeking, it does not provide an understanding of what sites cancer patients are most likely to access, e.g. official institutions, cancer charities, online forums for cancer patients, blogs, etc. This is important because we need to have a picture of what information sources cancer patients access online, and how these may differ in their trustworthiness. For example, lines 83-85, page 4, state that “websites’ information was often incomplete”, but there is no detail on what kind of websites these were. If such information is not included in the original studies, the authors of the present review need to say so and add some reflection on the shortcomings of past research.
The Introduction could also benefit from a distinction between trust in online cancer information vs. trust in the Internet, as the former refers to content and the latter, to a channel of information diffusion. Clarifying this aspect would help refine the goal of the present review in lines 116-118.
Another aspect which is underdeveloped is the definition of trust for the purpose of the present review (lines 99-100). The authors need to elaborate on the process of which trust is the outcome, i.e. is this process underpinned by heuristics or considered judgements, or both? And how might we expect heuristics or deliberate analysis to occur in relation to different websites? It may also need to be acknowledged that trust can be subjective and thus fulfil a psychological function (coping with threat). Cancer patients may be in a vulnerable state therefore their willingness to place trust in certain online information sources may be linked to their psychological need to reduce their vulnerability.
Line 221 – in Lussiez et al.’s study, the participants were thoracic surgery patients, so they are not patients with any type of cancer as the authors state.
In the Results, the section on cancer patients’ trust in online cancer information should have included information on how trust was measured in the 7 reviewed studies, i.e. the wording of the questions/scales used.


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

Input:
Title: THE EXTENT TO WHICH CANCER PATIENTS TRUST IN CANCER-RELATED ONLINE INFORMATION: A SYSTEMATIC REVIEW
Review round: 2
Reviewer: 1

Output:
Basic reporting: I commend the authors for improving this manuscript which I strongly recommend for publication. The manuscript reads much better now and makes a significant contribution to the field, as there is currently a lack of studies which have examined trust in online health information among cancer patients.
Experimental design: I am not familiar with RTI item bank and therefore cannot comment on it. However, I am convinced that the systematic review and appraisal of the studies included were conducted thoroughly.
It is good that the authors deleted the concept of 'active trust' from their manuscript as this term has very little scientific usage (I have never encountered 'active trust' in my own research on trust in online health information).
The authors provide a very clear distinction between trust and credibility and clarify the processes through which cancer patients can evaluate the credibility of online sources (heuristically or systematically) and place trust in them.
Validity of the findings: I think the findings are valid and the conclusions are pertinent. It would have been good if the authors could have included qualitative studies exploring why cancer patients trust or do not trust online health / cancer information, but this systematic review is nonetheless good. The number of studies addressing trust in online health information among cancer patients is relatively small so the authors did well to find any and to synthesise their findings. The authors highlight some of the antecedents of trust in online health information and also highlight the fact that the way researchers measure trust in online health information needs to be improved and made more transparent when reporting research findings. Last but not least, the authors include very good reflection on the strengths and limitations of their systematic review.
Additional comments: In the manuscript the authors provide much needed new insights into an area which is currently under-researched. I will follow with interest the authors' next research and publications and wish them good luck in their new research initiatives.


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: COMPUTATIONAL ANALYSIS OF FUNCTIONAL SNPS IN ALZHEIMER’S DISEASE-ASSOCIATED ENDOCYTOSIS GENES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Level of English is high. I've found just a few typos:
Line 29:  were further analyzed → was further analyzed
Line 39: may implicated → may be implicated
Line 46: with prevalence → with a prevalence of
Line 109: to have higher impact on → to have a higher impact on
Table 4: Headline is doubled (first two rows are the same).

Literature references are sufficient with an exception of linking R258Q variant to PD.(further explained in section 3).

It’s well-known now that there is no “junk DNA” and variants outside protein-coding regions might play a crucial role in many diseases. However, referring to the certain numbers (line 111; 1.26% and 1.46% for sSNPs & nsSNPs) seems somehow redundant & misleading without further explanation. It differs a lot between types of diseases (mendelian versus complex) and such quantification is very tricky. I would recommend rewriting this part with using more recent sources (again, numbers are not necessary).

In general, I would appreciate to better connect the general explanation of types of variants and their causal disease-causing effect with AD, not with chronic hepatitis B infection or familial dysautonomia (paragraph between lines 118-125).
Experimental design: I would like to see a better explanation of why the authors decided to limit their analysis to SYNJ1, PICALM and SH3KBP1. Late-Onset AD risk endocytosis-related genes involve also some other ones, e.g, BIN1, CD2AP, EPHA1, and SORL1.

The Methods section is clearly written. I’ve carefully reviewed the selection & settings of all predictors: authors seem to follow recommended settings & best practices or set their own ones in a very reasonable way. All the necessary details are provided.
Validity of the findings: The authors did not report that one of 56 highlighted variants (R258Q in SYNJ1) is strongly associated with Early-onset Parkinson Disease [PMID: 28331029]. The disease-causing effect is explained by the loss of dephosphorylation activity of Sac1 domain. It’s encouraging that this literature-validated variant introduced the highest total free energy in authors' simulations and keeps a promise that the other variants might be good targets for further investigation as well. The authors should check literature and discuss this variant properly in the manuscript.

Regarding the validation of highlighted variants in large cohorts from sequencing: I suggest to include also AD-control MSBB cohort (part of AMP-AD) – getting access is relatively easy and can help to pre-validate more variants (optional):
https://www.nature.com/articles/sdata2018185 (follow links to Synapse data as Data Citations)
Additional comments: The manuscript is clearly written and the topic is of high importance. In my opinion, the results are worth publishing. There are several areas, where I would suggest some improvements.


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: COMPUTATIONAL ANALYSIS OF FUNCTIONAL SNPS IN ALZHEIMER’S DISEASE-ASSOCIATED ENDOCYTOSIS GENES
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall satisfactory but with deficits in context integration with this study.

- Clear and unambiguous, professional English used throughout. Some grammatical and syntactical errors are found dispersed in the text, so authors may want to go over the document one more time, examples provided below:
L266: disorder -> disorders
L492: involve -> be involved
Table4_L6: (continue) -> (continued)

- Literature references, sufficient field background/context provided. This is where I feel the manuscript requires significant improvement as follows:
1) Regarding the proteins chosen for the study, in the introduction, it should be more clearly stated that the 3 proteins that were selected were the ones that either classified as “Validated risk factor” or “Interacts with validated risk factor…” in PMID: 22033521
2) Regarding SYNJ1, since this study tries to relate the results with AD, I feel that the authors need to improve the introduction, results and discussion section to address the following issues: Although in PMID: 22033521 SYNJ1 is a yeast Aβ suppressor, in a series of studies from the Cai group (PMID: 24052255, 26372964, 28900205) some of which are cited by the authors, it seems that in rodents SYNJ1 may be an Aβ enhancer or may increase pTau especially secondary to certain stress conditions such as TBI. In addition, mutations that selectively abolish Sac1 function lead to early onset parkinsonism whereas critical reduction of the dual phosphatase activity may be linked with neurodegeneration (maybe it would be helpful for authors to read PMID:27435091 for context). All these studies have to be discussed because they affect the definition of deleterious mutations in the context of AD. Authors to ensure that similar issues are not taking place with PICALM and SH3KBP1
3) L332-4: Please provide reference supporting the statement that sSNPs can result in protein misfolding
4) L363-4: Please explain why this is important “Furthermore, a portion… …in AD patients.”

- Professional article structure, figures, tables. Raw data shared: Good except minor inconsistency: either put “table… (continued)” in all tables that span multiple pages or in none.

- Self-contained with relevant results to hypotheses: Yes
Experimental design: Study is very good in this regard; minor improvement suggestions are given below.

- Original primary research within Aims and Scope of the journal: Yes

- Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap: Yes

- Rigorous investigation performed to a high technical & ethical standard: Yes

- Methods described with sufficient detail & information to replicate: Clearly written with all information required, except as below:
1) L438-449: The authors have included a nice validation for their approach. On the other hand, authors should clearly describe how they selected the 10 PSEN1 nsSNPs for AD. There are literally hundreds more and a lot of them have been more conclusively linked with AD (e.g. E280A to name one). Similarly, how they selected the random 5 not pathogenic. If applicable, the authors should consider including the rs63750592, rs63750771, and rs112451138 as they have the highest level of confidence of not being pathogenic.
2) L280-3 Authors mention critical role of miRNAs for regulating gene expression via 3’UTR but they haven’t performed a miRNA binding site prediction specific analysis (with e.g. TargetScan) for these genes. Maybe authors could comment that such an analysis was outside the scope of this study in the discussion.
Validity of the findings: There is good support for findings

- Data is robust, statistically sound, & controlled: Yes

- Conclusions are well stated, linked to original research question & limited to supporting results: Yes except as for SYNJ1 as related to the context described above.
Additional comments: The manuscript is clearly written, and it can be a good resource for the potential role of SNPs for these genes that have been implicated in AD. As, I have mentioned above the biggest deficits are in presenting findings in the context of existing literature and in relevance with AD.


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: COMPUTATIONAL ANALYSIS OF FUNCTIONAL SNPS IN ALZHEIMER’S DISEASE-ASSOCIATED ENDOCYTOSIS GENES
Review round: 2
Reviewer: 1

Output:
Basic reporting: Language has been significantly improved compared to the previous version but minor mistakes were introduced in the newly added parts. These could be addressed by the editorial team (no need for an additional round of revision).
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Thank you for addressing my concerns in the revised 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: INCIDENCE AND RISK OF DIALYSIS THERAPY WITHIN 30 DAYS AFTER CONTRAST ENHANCED COMPUTED TOMOGRAPHY IN PATIENTS CODED WITH CHRONIC KIDNEY DISEASE: A NATION-WIDE, POPULATION-BASED STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: -	Minor language improvements are necessary and rephrasing should be considered for example in lines 48, 66, 79-82 and 97.
-	Your introduction should explain some more relevant aspects that are important for the reader. Please comment briefly on the pathophysiology of CI-AKI. Please comment on the role of contrast media type (osmolarity), volume and application modality (intraarterial (angiography) vs. intravenous) in your introduction. You might refer to: Mayo Clin Proc. 2012 Apr;87(4):390-402. doi: 10.1016/j.mayocp.2012.01.012.
-	Thank you for providing the raw data. Please check that tables and figures are explained sufficiently in the accompanying text. E.g.:
o	Table 2: You should add definitions of the subgroups. Especially “advanced CKD subgroup” is not understandable by itself.
o	Figure 1 & 2: Please add a subtext to describe the figures and explain the abbreviations used.
Experimental design: -	Considering the chosen methodology (data extraction out of a national research database), the cohort size is deficient.
Validity of the findings: -	Please discuss your finding, that the incidence of CM-AKI is more frequent in CT without ICM (even though not statistically significant), in more detail. According to table 1, the population of CT with ICM was significantly younger and has significantly less comorbidities. Can you explain this? You should pick this up in your discussion.
-	In your discussion you mention the lack of information concerning the used contrast media type and volume. Please discuss this limitation of your study in more detail and refer to studies comparing isoosmolar and hypoosmolar contrast media. See Ann Intern Med. 2016 Mar 15;164(6):417-24. doi: 10.7326/M15-1402. Epub 2016 Feb 2.
-	Do you think the higher rate of AKI in the group of non-contrast CT could be because of “background fluctuation” of kidney function? See AJR Am J Roentgenol. 2009 Mar;192(3):711-8. doi: 10.2214/AJR.08.1413.
-	Please give a final statement about the significance of your work in the context of this relevant topic that is discussed so contradictorily. Which problems should be addressed in future studies and how should these be performed?
Additional comments: Thank you for your interesting manuscript about this relevant and controversially discussed topic. In my opinion, the main difficulty of your work is the relatively small cohort size in consideration of your experimental design. This raises significant statistical inaccuracy because of deficient adjusting. This needs to be discussed straightforwardly, as mentioned above.


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

Input:
Title: INCIDENCE AND RISK OF DIALYSIS THERAPY WITHIN 30 DAYS AFTER CONTRAST ENHANCED COMPUTED TOMOGRAPHY IN PATIENTS CODED WITH CHRONIC KIDNEY DISEASE: A NATION-WIDE, POPULATION-BASED STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: This manuscript by Shih Y et al examined the risk of dialysis therapy within 30 days of contrast enhanced CT in CKD patients. Overall, this study showed the dialysis and death risks within 30 days after CT scan in CKD patients are not increased irrespective of ICM.
Experimental design: Author collected data between 2012 and 2013. The CKD patients were divided into two groups. 1. 480 patients were CT with ICM, 2. 924 patients were CT without ICM. Outcome were measured using Kaplan-Meier survival analysis method and Cox proportional hazards model.
Validity of the findings: The rational and results are appreciated. It is also interesting to see in advanced CKD and ICU patients had no difference CT with or without.
Additional comments: 1.	How did the patients selected in each group? Randomly?
2.	Table 1: Add seperate p-value for each age group.
3.	There many study including large trials showing ICM leads to AKD. How could author justify beneficial and deleterious effect? How do they differ?
4.	Is there difference in ethnicity or any other parameter matters?
5.	Why within 30 days of CT? Is there any reason behind it?
6.	Data set and analysis are convincing but based on previous study reported and even our own clinic patients who undergoes CT with ICM get AKI.
7.	Method should be improved. How did data been collected
8.	All the patients are over 60 years of age. Do they matter to adjust the age for cumulative incidence of dialysis events?


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

Input:
Title: INCIDENCE AND RISK OF DIALYSIS THERAPY WITHIN 30 DAYS AFTER CONTRAST ENHANCED COMPUTED TOMOGRAPHY IN PATIENTS CODED WITH CHRONIC KIDNEY DISEASE: A NATION-WIDE, POPULATION-BASED STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: As the article is of particular impact and up-to-date, you might consider to cite the recent review by Mehran et al in NEJM (N Engl J Med 2019; 380:2146-2155 )DOI: 10.1056/NEJMra1805256 )
Experimental design: You might consider discussing your major limitation (small cohort size), which can not be improved due to the chosen methods, more critically.
Validity of the findings: no comment
Additional comments: Thank you for your extensive revision work. Your manuscript has improved considerably now. In my eyes, your study has one major limitation, which is the small cohort size. As this limitation can not be resolved, critical discussion is essential.


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

Input:
Title: INCIDENCE AND RISK OF DIALYSIS THERAPY WITHIN 30 DAYS AFTER CONTRAST ENHANCED COMPUTED TOMOGRAPHY IN PATIENTS CODED WITH CHRONIC KIDNEY DISEASE: A NATION-WIDE, POPULATION-BASED STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: 'no comment'
Experimental design: 'no comment'
Validity of the findings: 'no comment'
Additional comments: The author addressed all the queries I have raised on 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: INCIDENCE AND RISK OF DIALYSIS THERAPY WITHIN 30 DAYS AFTER CONTRAST ENHANCED COMPUTED TOMOGRAPHY IN PATIENTS CODED WITH CHRONIC KIDNEY DISEASE: A NATION-WIDE, POPULATION-BASED STUDY
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Thank you for your revision. The manuscript has improved markedly and the limitations of the study are now sufficiently 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: QPCR AND LOOP MEDIATED ISOTHERMAL AMPLIFICATION FOR RAPID DETECTION OF USTILAGO TRITICI
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this article Yan et al. attempted to describe novel approaches for rapid detection of Ustilago tritici by qPCR and loop mediated isothermal amplification methods. However, obtained data in the current manuscript were a preliminary screening of several factors and trials in order to get an initial outline of how designed primers work in vitro conditions. No description of scientific hypothesis driven to or how to achieve those unique and specific primers was shown in this research. Importantly, no practical experiments were conducted to examine those methods with samples from the fields. Other than that, the use of English in this manuscript was poor and did not meet the professional standard for publishing as a scientific research article. Therefore, I do not think the current version of the article is worth for publication in this journal.

The introduction was unfocused, particularly lacking a description of wheat loose smut’s pathogen, its life cycle, as well as which candidate factors that current approaches are targeted to establish a diagnostic strategy.

The supplemental files of this manuscript were impossible to access. There may be an error from the journal system I would say.

Not only tables of primers (Table 1 as in line 98 and Table 2 as in line 101) were absence, how distinct approaches to design as well as how unique of the designed primers compared to previous publications were also undescribed in this manuscript.

There was no data throughout manuscript representing for the statement in line 27-29 of abstract “We successfully performed qPCR and LAMP assays on two wheat loose smut wheat samples, and conﬁrmed sequenced U. tritici infection by subsequently sequencing”.
Experimental design: More experiments must be conducted to address these key questions:

1. What kind of wheat loose smut samples (soil, wheat stems, seeds, etc.) in fields would be ideal targets for these diagnostic methods?

2. How these methods could be applied to the field samples? Directly running the raw samples or performing with isolated DNA from the samples?

3. It may be a very big question, but let’s think about how many percent or positive obtained results would be the threshold line for a conclusion of a disease outbreak or a need to be controlled?
Validity of the findings: The data obtained in vitro by isolated DNA screening would be generally different from raw field samples. Therefore, based on these preliminary data, additional experiments applied in field samples must be provided to demonstrate the feasibility of the methods.
Additional comments: The manuscript must be revised by not only a senior scientist but also a native speaker to obtain professional criteria of an original research article.


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: QPCR AND LOOP MEDIATED ISOTHERMAL AMPLIFICATION FOR RAPID DETECTION OF USTILAGO TRITICI
Review round: 1
Reviewer: 2

Output:
Basic reporting: Line 3 & 4: replace “/” with comma
Line 18: Please add seed-borne fungal disease in the first sentence
Line 20: replace “were” with are and “specify” with specificity
Line 21: correct the sentence “were carried out” to is assessed by using genomic DNA of …..
Line 25: Please make the primer nomenclatures consistent throughout the manuscript. Like L-139 vs. L139
Line 29: replace “established” to establishes
Line 30: replace “could” to “can” and diagnose to “diagnosis.”
Line 34-38: Authors did not mention the pathogenesis of the wheat loose smut disease. Two or three sentences for the introduction of the disease and a few sentences for the necessity of the diagnostic test would help the reader get a gist of the manuscript.
Lane 42: incomplete sentence “single used…”
Line 81: replace “bacterial strain” with Fungal strains
Line 83: provide the source of fungi
Line 85: replace “24 h” to 24 hours (h)
Line 86: sentence unclear. Please replace “12h days …..” to for the 12h of the day and 18deg C,  12h of the night.
Line 86 and 87: provide the source of U. tritici and wheat powdery mildew. A supplementary figure with the phenotypical observations of the fungi will be very informative for the readers.
Line 88: Provide the source of PDA media
Line 91: please cite CTAB method and introduce the acronym
Line 99: please cite articles publishing the DNA sequences. Which genome sequences were chosen for the qPCR and LAMP assay? and why? Are there any reference genes or housekeeping genes selected as a baseline for the qPCR data analysis?
Line 103: replace “QPCR” to “qPCR”
Line 105: replace “are……” with is used to detect the specificity of the primers.
Line 105: Provide the details of the thermocycler used.
Line 118: replace “QPCR” to “qPCR”
Line 159: replace “there for” with therefore
Line 192: please confirm qPCR primer Y334 or  Y430
Line 205: correct the sentence both……
Figure legends: please change all the fungal species names to the italic font.
Experimental design: Authors approach to develop a specific, rapid, and accurate DNA-based diagnostic test for the wheat loose smut is encouraging. However, the diagnostic test lacks robust experimental data to support the findings and cost-analysis.
Validity of the findings: There is a gap in the explanation of how the detection of an organism can help prevent the disease and why it is so essential to distinguish a species. Authors can discuss these topics in the introduction section. Authors did not explain the strategy in designing the primer set. If the genome sequence of the U. tritici is annotated, then please present and explain the goal behind the amplification of this specific region in the genome. Also, the authors may include a fungal housekeeping gene as a reference gene to confirm relative amplification of the target gene.
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: PHYSIOLOGICAL AND ANATOMICAL INVESTIGATION OF THE AUDITORY BRAINSTEM IN THE FAT-TAILED DUNNART (SMINTHOPSIS CRASSICAUDATA)
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The article by Garrett et al. describes auditory brainstem responses (ABR) in the anaesthetized fat-tailed dunnart, with a surprising finding of lower thresholds for higher frequencies towards 50kHz. Unfortunately, higher frequencies are not examined due to technical limitations, and therefore peak sensitivities could not be tested. Although a more complete characterization would be needed to answer this, the present article discusses these limitations appropriately, and within a well-framed comparative context. Next, the authors present a detailed cytoarchitectural description of brainstem auditory nuclei in this species using Nissl staining. In my opinion, this is a timely and well-written article that describes some very basic features of dunnart auditory neurobiology, particularly relevant due to the scarcity of data in dasyurid marsupials.

I only have minor comments for the authors to address before proceeding to publication:

-	Use italics or underline for all species’ scientific names (lines 69-72).
-	There’s a space missing in lines 81 and 104.
-	Do not use capital letters for common names or reagents (e.g. fat-tailed dunnart in title; ketamine, cresyl violet, etc)
-	Fig. 1 title says ‘d’at-tailed
-	Figs 3 and 4 should clearly indicate the plane of section in the result section and figure legends (at least coordinates such as 'lateral', 'anterior' to ease spatial comparisons). Including a schematics of the approximate region where sections were taken would be great in both Fig. 3 and 4.
-	Improve the labelling of the nuclei of interest in Figs. 3 and 4, for example by outlining all regions and/or placing the lettering at the centre of each structure, or outside with a line pointing to them, to improve clarity and flow with 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: PHYSIOLOGICAL AND ANATOMICAL INVESTIGATION OF THE AUDITORY BRAINSTEM IN THE FAT-TAILED DUNNART (SMINTHOPSIS CRASSICAUDATA)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The present study investigated the auditory system of the marsupial fat-tailed Dunnart (Sminthopsis crassicaudata), because auditory brainstem of this species is not investigated in detail so far.
Auditory brainstem responses were recorded showing similar response to other mammalian species, however, the dunnart is most sensitive to higher frequencies increasing up to 47.5 kHz. Morphological investigation of auditory brainstem by use of Nissl-staining  revealed auditory nuclei (cochlear nerve root nucleus, cochlear nucleus subdivisions and nuclei of the superior olivary complex) comparable to that in other mammalian/marsupial species.
The manuscript in general is well written, however a more detailed anatomical/histochemical analysis of auditory brainstem nuclei would be desirable. The manuscript should be improved as suggested in general comments for the author.
Experimental design: The research question is clearly defined and would be of interest to know whether auditory brainstem organization in fat-tailed Dunnart is similar or different to that in eutherian or other metatherian species with respect to range of frequency hearing.
Methods are described with sufficient Information to be reproducible. Still, histological analysis of auditory brainstem nuclei can be improved by a more detailed Investigation of auditory nuclei (subdivisions, size, extent, neuron types, myeloarchitecture).
Validity of the findings: Validity of findings is given, but can be improved by a more detailed morphological characterization of auditory nuclei (cochlear nucleus with each subregion, single nuclei of the superior olilvary complex) supporting physiological data.
Additional comments: Introduction
Page 3-4: The auditory brainstem nuclei of marsupials display distinct features compared to eutherian mammals, this should be more clarified in the introduction.
Results
Page 8, line 199: A more sparsely populated PVCN containing larger nuclei was observed in more caudally located sections. Please clarify this statement, what is think about “containing larger nuclei” in more caudally located sections (Figure 3c,d,e,f)?
Page 8, line 195, line 202, line 209: Literature should not to be included into the results, but are a part of discussion.
Page 8: line 194-200: In marsupials the partition of the CN is somewhat different from that of the eutherian mammals: anterolateral and posteromedial AVCN as well as lateral PVCN and octopus cell area are named and should be also identified in the here investigated marsupial.
Page 8, line 201: At the beginning of the DCN, the DCN appears not to be tri-laminar (Figure 3b, d)?! Clearly lamination was only seen in Fig. 3f. The correct designations of the DCN layers are as follows beginning with the outermost layer: molecular layer, fusiform layer and polymorphic layer (see also Bazwinsky-Wutschke et al. 2008).
Page 9, line 221, line 223: Avoid the involvement of citations in the result section.
The description of the LSO is very sparsely. According to other marsupials, the drop-shaped LSO can be subdivided into a central and marginal region (see also Willard and Martin 1983). Do you have seen the SPN (superior paraolivary nucleus) in the SOC?
To improve and to deepen the histological analysis, in general the rostro-caudal extent of each auditory brainstem nucleus can be analyzed and features described.
Discussion
Page 11: The MNTB-LSO system should be more discussed with the background that the MNTB is the major source of acoustically evoked glycinergic inhibition within SOC and a key nucleus in the sound localization (transforming the bushy cell excitatory input into inhibitory output acting on the ipsilateral LSO, MSO and SPN).


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: PHYSIOLOGICAL AND ANATOMICAL INVESTIGATION OF THE AUDITORY BRAINSTEM IN THE FAT-TAILED DUNNART (SMINTHOPSIS CRASSICAUDATA)
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article was well written, clear and concise.
Literature citations were generally appropriate.
Article was structured well.
Experimental design: Experimental design followed prior methods, so I have no issues.
Research question was well defined.
Validity of the findings: The methods used were tried-and-true.  Thus, I have confidence that the findings and conclusions are valid.
Additional comments: What follows are just some general comments and questions that the authors might consider.
Ln 105.  Was sex not known?
Ln 114. Some additional details regarding the ABR measurements would be helpful.  How was the system calibrated?  How was SPL of the stimulus measured?  In the ear canal? Or simply the output of the earphone?
Ln 143.  Visual estimation of ABR threshold can be quite variable.  To give the reader some sense of what was done, could you provide the estimate of the threshold for the example ABRs shown in Figure 2a?


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: FOXO1 IS A CRITICAL REGULATOR OF HEPATOCYTE LIPID DEPOSITION IN CHRONIC STRESS MICE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The use of English is clear, though there are plenty of grammar mistakes await the author to correct. Introduction and background are included, and structure seems in discipline. Figure 1A and 6A doesn’t seem qualified for publication, and there are errors in legend of these two figures. Raw data supplied.
Experimental design: It is an original primary research within scope of the journal. Methods are all sufficiently described, and conducted in conformity with the prevailing ethical standards in the field. The scientific hypothesis of this article is clearly proposed, and the question is quite meaningful, however, the elucidation may be incomprehensive.
Validity of the findings: Conclusions are well stated, linked to original research question，and limited to supporting results.
Additional comments: [1] The most important issue of this article is that although you claimed to bring the molecular mechanisms of stress-induced lipid metabolism disorders some clarity, however, the design of the whole experiment only explore the result of inhibiting FoxO1, which is hard to call a mechanism.

[2] The language and the style of writing certainly need improvement. The abstract should be a synopsis, which means you should summarize everything rather than present what and why  you did in simple sentences. Similar language problems happen in the result section, where logic between each figure should be slightly elucidated, or readers may be easily lost track of your purpose. Furthermore, the order you present your result might need to be in line with how you put it in discussion, in this way you can bring unification into your article. Obvious grammar mistakes can be found, please correct them all before publication.

[3] The figures 1A and 6A can hardly be considered qualified. I checked the original pictures in raw data, still, the shadows and dots can be a real problem for analyzing, and the actin in 6A doesn’t seem like it’s on a straight line.

[4] NAFLD is a major risk factor for the development of type 2 diabetes and a central feature of the metabolic syndrome(Asrih & Jornayvaz, 2013). There are literatures featured in FoxO1, adipocyte differentiation, and its regulation by insulin(Puigserver et al., 2003)(Nakae et al., 2003), and certain mechanisms are presented. However, such progresses are not mentioned well enough in this article, except in line 247 and ITT. However, in line 247, that’s just an introduction of FoxO1, and ITT can merely explain a mechanism. Clearing this may help valid your results.

[5] The introduction of corticosterone is abrupt, and if you want to call it a model, please verify it or provide with previous studies. And your corticosterone treatment was only applied to a cell line, but if you want to discuss the HPA axis, in vivo treatment may be better.

[6] In your methods, you mention that your data was mainly analyzed using one-way ANOVA. However, you are using corticosterone and as1842856 treatments, and they are two different effectors, thus the statistical approach should be two-way ANOVA. The significance may alter with the change of analyzing methods, please change it in modification.

[7] Titles of figures are overly narrative, please summarize your theme of each figure.


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: FOXO1 IS A CRITICAL REGULATOR OF HEPATOCYTE LIPID DEPOSITION IN CHRONIC STRESS MICE
Review round: 1
Reviewer: 2

Output:
Basic reporting: In this paper, Liu and colleagues report that FoxO1 is a critical regulator of hepatocyte lipid deposition in chronic stress mice, extending their previous findings. The results are interesting and the article is valuable for publication. However, some small concerns need to be addressed.
1) Use consistent abbreviations in the manuscript, e.g. TG for triglyceride, FFA for free fatty acid (FFA), etc.
2) It’s better to include all the used reagents into Section “Reagents and Antibodies” in Materials & Methods.
3) Line 78: The as1842856 was administered by gavage, whether this kind of treatment causes additional stress to the animals?
4) Line 124: Check “Bonferroni’s multi-ple comparisons”.
5) Line 188: “After treated with 1 μm corticosterone for 48 h”. Does it mean 1 μM?
6) Line 260-263: Include the reference.
7) The authors mentioned that Other synthetically related genes, ACC1 and SCD1 showed no significant difference between the four groups, it’s better to give one or two sentences to discuss this observation.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: None.


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

Input:
Title: FOXO1 IS A CRITICAL REGULATOR OF HEPATOCYTE LIPID DEPOSITION IN CHRONIC STRESS MICE
Review round: 1
Reviewer: 3

Output:
Basic reporting: English needs polishing furhter.
Experimental design: This paper raised a good question, the experimental design is OK, but the data set are kind of worrysome.
Validity of the findings: See the specific comments to author below.
Additional comments: Liu et al. in their manuscript entitled “Foxo1 is a critical regulatory of hepatocyte lipid deposition in chronic stress mice” tried to come up with a mechanistic explanation for how Foxo1 regulates chronic stress-induced liver lipid deposition, a basis for the nonalchoholic fatty liver diseases (NAFLD). It is potentially interesting. The data set need to be improved and the writing need more editing and enhancement though.

Specific comments:

1. Does as1842856 has any effect on the morphology and function of the liver? In figure2A, the picture of con and con+as1842856 looks different.
2. Despite that as182856 does have effect on Foxo1 phosphorylation and target gene expression, it does not rescue or change the effect of stress condition on body weight loss at all (Figure B). This is an interesting puzzle and should be explained or speculated.
3. In results, the description on figure3A and 3B seems to be mistaken. Also in figure3B, the stress+as1842856 group is dramatically different from control, they are not consistent with what is described in the paper.
4. Why as1842856 has no effect on basal Foxo1 activity under steady condition?
5. The English language should be improved throughout the manusript.


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: FOXO1 IS A CRITICAL REGULATOR OF HEPATOCYTE LIPID DEPOSITION IN CHRONIC STRESS MICE
Review round: 2
Reviewer: 1

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


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

Input:
Title: COMPARISON OF INJURY PATTERN AND CLINICAL OUTCOMES BETWEEN YOUNG ADULTS AND ELDERLY PATIENTS WITH ALCOHOL-RELATED INJURY IN SOUTH KOREA 2011–2016
Review round: 1
Reviewer: 1

Output:
Basic reporting: This research study explores an area that is relevant to a cross-disciplinary readership and is of considerable relevance to clinical practice. It is coherent and succinct. The aims are clearly stated with structured data that is well organised. The results, discussion and limitations are clearly linked to the aims and methodology.
Experimental design: The methodology is robust, with a large data set interrogated to produce results that have sufficient power and significance to reduce the likelihood of false positive and negative results. Although limited by retrospective design and hence limited by ascertainment and observer bias, the sample size is representative, with the use of a control group against which to compare odds ratios of relative risk. Outcomes are valid and statistical analysis appropriate to experimental design.
Validity of the findings: The study is of high quality and fills a gap in knowledge surrounding the association between injury and alcohol use, with adequate reference to the literature,. It has considerable external validity, given that it draws on data obtained from clinical settings that could be conveniently replicated across other similar settings in other countries.
Additional comments: I found this paper of high quality, with robust aims, hypothesis, methodology, reporting of results and the discussion was balanced, qualified by limitations to the interpretation of results.


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

Input:
Title: COMPARISON OF INJURY PATTERN AND CLINICAL OUTCOMES BETWEEN YOUNG ADULTS AND ELDERLY PATIENTS WITH ALCOHOL-RELATED INJURY IN SOUTH KOREA 2011–2016
Review round: 1
Reviewer: 2

Output:
Basic reporting: The first paragraph ie the abstract part needs to be a synopsis of the paper ie defining and improving to make it more succinct and accurate:In the first paragraph, background rick should be risk. Not really understanding the methodology ie sentence:but those who visited the ED 48 h after injury, with unknown clinical outcomes.  I'm assuming this was a retrospective study.  Were the notes reviewed? Where did the information initially come from to get the patient group?

So I would suggest that the initial abstract is revised ie background and methods.

The above is clarified in the materials and methods in the main body of the paper. Was there any information about quantity of alcohol consumed?  Any audit C data?  This is still seen as the best tool for elderly alcohol consumption.  Was there any information collected about poly pharmacy?

For a future study may also be worth breaking down the over 65s into age brackets.

Probably worth referencing the below:
Use of blood alcohol concentration in resuscitation room patients.

Csipke E1, Touquet R, Patel T, Franklin J, Brown A, Holloway P, Batrick N, Crawford MJ.
Experimental design: No comment.  I thought the design was appropriate and any flaws were mentioned.
Validity of the findings: No comment
Additional comments: I would just suggesting revising the abstract part and perhaps reading the paper suggested to see if this could be added in somewhere.


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: AN UPDATE ON ANATOMY AND FUNCTION OF THE TELEOST OLFACTORY SYSTEM
Review round: 1
Reviewer: 1

Output:
Basic reporting: No Comment
Experimental design: a review: not applicable
Validity of the findings: a review
Additional comments: This review is an update of the excellent 2013 review (J. Chem. Ecol 39:451-464) by the two authors; however , the previous review dealt primarily with fish olfactory receptor neurons, their properties, projections and tuning; the present manuscript includes additional information covering olfactory receptors and higher order olfactory neurons.  Of the 100 citations listed in the current manuscript 72% (72 of the 100) were not included in the previous review; however, only 19 of the references cited were published over the past five years (i.e. since the publication in 2013 of the previous review by the present authors).  That only 19% of the cited literature provide new information on the topic does not strongly support the necessity of an “update” on the anatomy and function of the teleost olfactory system.  The primary message obtained from this “update” is that little new recent information (last five years) has been published on the topic. In spite of this deficiency, the present manuscript along with the suggested alterations will provide a succinct review on the anatomy and function of the olfactory systems in teleosts and be of interest to those who are not specialists in this field.

1.	Line(L) 7: single kind? What does this mean?  Some fish olfactory organs are arranged into various numbers of lamellae which splay out from a midline raphe, some others have an olfactory organ that contain lamellae but are organized differently (e.g. see Theisen et al. Mar Biol 110:127-135 (1991) Figs 2-4) and still others lack lamellae (Zeiske et al., Cell Tissue Res 172:246-267, 1976).
2.	L 88: water flow through fish olfactory organs can be propelled by fish movements, ciliary action of the non-olfactory cilia (in isosmates) and muscular pumping actions (in cyclosmates)-see Doving et al., Acta Zool 58:25-253, 1977 for definitions of cyclosmates and isosmates).
3.	L90: true? sniffing is absent (incorrect): see cyclosmates defined in the above Doving reference.
4.	L 91-92: Although teleosts lack a separate vomeronasal organ, a recent report (Biechi et al., Scientific Reports 7:44295, 2017) provided evidence for an accessory olfactory system in zebrafish;  previous reports in lungfish also indicate the existence of a VNO system (Gonzalez et al., Frontiers in Neuroanatomy 4, article 130, 2010; Nakamuta et al., Anat. Rec. 295:481-491, 2012).
5.	L 101-102: There are 4 anatomical types of fish olfactory receptor neurons well-identified, but there is also a fifth that has been reported: “onion-like” (from the Yoshihara lab).
6.	L105: “sustain”? Do you mean “contain”?
7.	L 150: “middle” should be “”medial”
8.	L155:regions indicated (omit “have”); medial olfactory bundle
9.	L157: “medial”; omit “would”; relayS
10.	Ls 166-179 and throughout the review: Authors seem to like past-perfect tense rather than simple past tense which contains fewer words; e.g. “have been”  better as “were”; “has been” better as “was”
11.	L 174 Golf: authors need to check the literature as I don’t think this particular G-protein has been demonstrated in fish.
12.	L205: (Sato & Suzuki, 2000); however, in seawater….
13.	Section titled” “The teleost telencephalon”-especially lines 212-244: The development of the teleost telencephalon is quite complicated with respect to “eversion” which greatly complicates the homologous relationships between telencephalic structures in teleosts vs other vertebrates.  A newer review of this situation was published in Neural Development 7, article #212 (2012) that I recommend the authors to review and determine whether their discussion needs updating/alterations especially in light of the authors’ title of this manuscript (e.g. “an update”).
14.	The paragraph starting with L 243 contains sentences that are too long;  I suggest the following: Omit :although: & start sentence with “The differences..and end the sentence with a period after vertebrates.  Start the next sentence with “Studies” and end the sentence with vertebrates.  Omit “while” and start the next sentence with Also, the Vs represents….; L 249: substantia innominate, which are related; L 250: omit “on the other hand”-start sentence with “In the dorsal area…; L 251: the pallium, whereas the most medial….end sentence with “amygdala”. L 252: The DI is likely homologous to the hippocampus and the Dp is the main…
15.	L 256: omit “allowed” established that the pallial regions and the subpallial nuclei (move all the abbreviations (here) receive projections from the olfactory bulb.  Start the next sentence with “The Vv region….
16.	L 302: omit “On the other hand”
17.	L 307-308: The statement depends on the number of components in a mixture (e.g. see Valentincic et al. J. Fish Biol 79:33-52, 2011).
18.	Ls 309—310: the statements concerning “decomposing and “de-correlating” odors” is not well-explained and may not be relevant to the identification of an odorant.  This concept is mostly correlated with work presented from the Friedrich & Laurent laboratories as a working hypothesis-however, see Nikonov & Caprio, J. Neurophysiol 98 (2007) p. 1916 “Response time consideration”.
18. L 327 & Figure 1: I recommend some caution when observing recorded olfactory waves especially within the beta-band frequencies and assuming the cause is synchronous neural activity (see Diaz et al. J. Neurosci 27(34): 9238-9245, 2007.


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: AN UPDATE ON ANATOMY AND FUNCTION OF THE TELEOST OLFACTORY SYSTEM
Review round: 1
Reviewer: 2

Output:
Basic reporting: The Review article entitled “An update on anatomy and function of the teleost olfactory
system” by Olivares and Schmachtenberg gives an good but rather short overview on the teleost olfactory system.

The Review article is very well written in a professional English, and it was a pleasure to read it. It is certainly of broad and cross-disciplinary interest and within the scope of the journal.

To my knowledge there are no recent reviews of the teleost olfactory system and I'm happy that the authors made the effort to write this review.

However, I feel that not all the relevant literature has been cited. I strongly encourage the authors to overwork the citations of the manuscript. Every single statement needs an appropriate citation/ reference. (Also see general comments to the authors.)

The unique figure is also not optimal and not very meaningful. I think that some more figures (transduction, general scheme of the fish olfactory system etc.) could substantially improve the manuscript, and make it easier for the potential readers to follow and understand the review.

For the rest, I find myself in the unusual position of not having major complaints. Some minor suggestions can be found in the "General comments to the authors" section.
Experimental design: Not much to complain here.
Validity of the findings: No comment.
Additional comments: Here some minor comments that should be addressed:

-some  references that really should be  included are:

1: Wanner AA, Genoud C, Friedrich RW. 3-dimensional electron microscopic imaging  of the zebrafish olfactory bulb and dense reconstruction of neurons. Sci Data. 2016 Nov 8;3:160100. doi: 10.1038/sdata.2016.100.

2: Wanner AA, Genoud C, Masudi T, Siksou L, Friedrich RW. Dense EM-based reconstruction of the interglomerular projectome in the zebrafish olfactory bulb. Nat Neurosci. 2016 Jun;19(6):816-25. doi: 10.1038/nn.4290.

3: Sato Y, Miyasaka N, Yoshihara Y. Mutually exclusive glomerular innervation by  two distinct types of olfactory sensory neurons revealed in transgenic zebrafish. J Neurosci. 2005 May 18;25(20):4889-97.

- In the abstract you speak about 5 types of olfactory receptor neurons. In the relevant part of the review you mention only 4 types. As far as I know, the fifth type of receptor neuron is the pear-shaped type described by Wakisaka et al., 2017. Please correct this incongruity.

- line 34: "....ranging from micromolar to nanomolar....". Here you should could be a little more specific.

- lines 46-48: You distinguish between "social odors" and "pheromonal odors".  What is the difference between social- and pheromonal odors? Please clarify or revise this passage.

- lines 64-70: In this passage of your review you should cite also the review of Hamdani el H and  Døving KB (The functional organization of the fish olfactory system).

- lines 73-74: What do you mean with "Four older research articles and 22 general articles on vertebrate olfaction were also included in the general data base." . Where do this older and  general articles come from?

-around line 110: Here you should consider adding a reference that describes the receptor neurons of terrestrial vertebrates.

-lines 118-131: Here some references are missing. Where did you get the information that you review in this section? Please add some adequate citations.

-line 152: "....chemotropic...." should most probably be "....chemotopic....".

-section "The odor transduction process": Here (or at some other place of thre review) you should mention the adenosine receptor A2c  and its transduction cascade described  by Wakisaka et al., 2017.

-lines 202-208: It would be great if you could explain a little better what the function of the calcium-activated chloride channel in seawater species is.

- Several times you compare the olfactory system of  aquatic and terrestrial vertebrates, but you do not mention the olfactory system of amphibians or other aquatic vertebrates/ tetrapods at all. You should maybe include some information about this.


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: INTERSPECIFIC VARIATION IN THE LIMB LONG BONES AMONG MODERN RHINOCEROSES—EXTENT AND DRIVERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: In the manuscript titled “Interspecific variation in the limb long bones among modern rhinos - extent and drivers”, Mallet and co-authors present a neat study on the long-bones of the upper limb in modern rhinoceroses using a three-dimensional geometric morphometric approach. However, considering the broad literature on limb morphology in relation to biotic and abiotic covariates (e.g. phylogeny, habitat etc.), I am surprised that this study lacks hypotheses on what may be found using a 3D-GM assessment of extant rhinocerotid proximal limb bones. I think this study is missing a clear and testable hypothesis; this, unfortunately, makes the Results section quite laborious to read, as it has no focal point to base the descriptions around. I suggest that even a very simple biogeographic, size-related or phylogeny-based hypothesis will allow the Results and Discussion sections to flow more smoothly towards a focused conclusion.
This article is in general well written and the points are put across in a logical manner. However, I have come across numerous examples of sentences which are in need of restructuring, and words used which are inappropriate (“pending” rather than “depending”, and possessive apostrophes being the chief culprits). There is also an odd combination of American and British English used – this is not a true criticism, but may be something which is in need of consistency throughout the article in accordance with the journal guidelines. I have chosen not to highlight every section in the article which I think is in need of adjustment, but I strongly recommend that the authors have this manuscript read over by a native English speaker before resubmission. The changes themselves will not be major, but there are many instances where a rearrangement of the sentence will allow a much better flow for the reader.
The tables, figures, references and supplementary information are well laid out, logical and easy to understand. There are a few references I feel could be added in key areas - I have made a few suggestions in the attached PDF file.
Experimental design: This article is well suited to the aims and scope of PeerJ; it represents an original research article within the biological sciences, performed to a high technical standard. The authors lay out several issues and knowledge gaps (e.g. the label of "graviportal" and whether rhinoceroses can be considered as such). The authors lay out their reasons for chosing rhinoceroses as a subject group well, with logical arguments. They then accumulated an excellent sample of modern rhinoceros upper limb bones, and have used more computationally intensive methods than were previously available to researchers studying locomotor morphology in these animals. The methodologies used are well laid out and informative, although missing a few details which I have highlighted and commented upon in the PDF. The results are comprehensive in their anatomical detail, if a little exhaustive to read at times. The discussion is good, with sound descriptions and comparisons. I feel the authors can delve deeper into the comparative aspects of different parts of the bone which they mention as key features, and several biomechanical concepts need to be readdressed to fit into the story that is being told. My main criticism of the article is the lack of specific aim and testable hypothesis – this absence bleeds down through the results and discussion sections, making them appear disjointed at times. Ecological, biomechanical and phylogenetic aspects are touched upon, but not really discussed in the scope of Rhinocerotidae evolutionary history. I would like to see more engagement with previous literature sources both recent and classic, and I strongly recommend large swathes of results section be relegated to the supplement to benefit whatever hypothesis the authors choose to go with.
Validity of the findings: The findings of this study most definitely benefit the wider literature. The results for interspecific differences between the different rhinoceros species will be valuable for any future studies based on rhinoceros locomotion, graviportality, or general perissodactyl biology. All data utilised for the study have been provided for the reviewers in well organised supplements or ancillary table and figure files, and from what I can tell the methodologies are robust, statistically sound and controlled. I do have some thoughts about the effect of high dimensionality one the data, and how this might affect the statistical comparisons, but these are minor concerns.
Unfortunately, as there was no specific research question or hypothesis, it is difficult to gague the conclusions. Yes, they are limited to supporting the results presented with no rampant speculation. However, I believe that there are aspects of the results coming from this article which would benefit from informed speculation, without drawing concrete conclusions but positing ideas which could be tested in future studies. I have included suggestions within the attached PDF. With the addition of a testable hypothesis and specific research aim, the results will be easier to interpret, the discussion will be more focused, and the conclusions will benefit.
Additional comments: I applaud the amount of work that has gone into this study, and on the whole I am a big fan of this work. The set-up for examining rhinoceros locomotor morphology in the framework of adaptations for “graviportal” locomotion is, in my opinion, the best place to start - of all the modern “graviportal” mammals, rhinos are perhaps most contentiously labelled so. The sample you have acrued is impressive, and clearly a lot of hard work went into the scanning and landmarking procedures.

Given the title and setup to the article, I had the biggest issue with the results section. The reason is simple - there was no actual reference to the shape of the bones of the species for well over 2/3 of the results. Theoretical shapes at axis minima and maxima are excellent means by which to visually inspect what aspects of morphology are most highly variable in a sample. However, the premise of this article (as I understood it) was to inspect the differences between the five extant taxa. There is not much reporting of that for much of the results, but rather an exhaustive report of what features are shaped differently along each PC axis. That, by itself, is interesting - I am not taking anything away from the work put in to identifying every feature which varies. However, I think that in an article quantifying "interspecific variation in the limb long bones of modern rhinos", I expected comparative results between the species to be the centre-piece of the results. Even as a comparative anatomist, I found reading the first two thirds of the results section quite tough, until the interspecific comparisons section. This issue, I think, can be tied back to lack of testable hypothesis – what question are the authors asking, what did they anticipate to be the answer, and do the data support that hypothesis? Once the hypothesis is decided upon, I think the text will fall into place quite easily, and discussion points/speculation will become easier to justify.

With all that being said, I am of the firm belief that this work is an ideal launchpad from which to test theories on rhinoceros locomotion, and the relationship between size and shape of the appendicular skeleton in so called “graviportal” mammals. Beyond that, it is a valuable window on the quite surprising morphological variation exhibited by the five modern rhinoceros species, a group which have for many years had their postcranial morphology somewhat maligned. I would very much like to see this study published, and I hope that the suggestions I have offered may help it along the way.

I wish the authors the best of luck with this study, however they chose to proceed.

Kind regards,
Dr. Jamie MacLaren


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: INTERSPECIFIC VARIATION IN THE LIMB LONG BONES AMONG MODERN RHINOCEROSES—EXTENT AND DRIVERS
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper is, for the most part, clear and unambiguous. Each section is well-delineated. The introduction hits the major points of appropriate background for the reader. Methods are not completely described in 'Landmark Digitization' (see attached PDF). Some of the technical definitions and wording is lacking at present as well. Figures are well organized and captioned, and only require a few changes. The raw data is available. Hypothesis is vague. This is more of an exploratory study, but the results, implications, and conclusions are strong.
Experimental design: This paper uses established methods (PCA, sliding semi-landmarks, generalized procrustes analysis, etc.) for a novel system: limb bones of extant rhinos. The methods are sound for the most part (see details in attached document), but lack sufficient detail to replicate. The PCA analysis is clear, but the sliding semi-landmark, the Procrustes superimposition with a spline optimized step, and the use of centroid size needs to be expanded upon. This was the main reason I chose 'minor revision' for the paper.

This paper fills an existing knowledge gap because extant rhino limb bones have never been quantified using such an extensive landmark analysis and with such a large sample size. This paper is an important addition to the limited quantitative morphological analyses of rhinocerotidae.
Validity of the findings: The results are valid and meaningful. The paper opts for a dual statistical and descriptive approach. While there are no clear hypotheses, the conclusions are well stated and link strongly with the introduction. The paper makes clear what is data and what is speculation.

All raw data is provided, but there is no associated R Code. While the script that was used to run the analyses is not required, it would be extremely helpful for data replication. See attached.
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: INTERSPECIFIC VARIATION IN THE LIMB LONG BONES AMONG MODERN RHINOCEROSES—EXTENT AND DRIVERS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors present a 3-D morphometric study of the limb bones, specifically the long bones, of extant rhinoceroses. They examine the variation among their samples from the five extant rhinoceros species to assess the effect of body size (using centroid size from their data and reported average body masses for each species) and phylogeny. Their results generally show that there is a strong phylogenetic signal, separating three groups: African rhinos, Rhinoceros, and Dicerorhinus. They also record an effect of body size that interestingly is most pronounced for the radius and ulna. The authors use these data to draw conclusions about the evolution of large body size in rhinoceroses and how rhinocerotid adaptations for large body size compare to those of other large terrestrial mammals. Cursory comparisons with hippopotamuses and elephants suggest different types of adaptations in rhinos as compared to these groups.

Overall, this is an interesting study that provides a quantitative approach to examining variation in bone shape that can be related to phylogeny, body size, and locomotion. With the amount of attention that has been given to the morphology of horses, it is refreshing to see current studies like this one and those of MacLaren and colleagues examining the other members of the Perissodactyla. I am particularly hoping that this study will be followed by examination of interspecific variation in other rhinocerotoids. I recommend publication with minor revision.

Listed below are suggested edits, with numbers referring to lines in the manuscript. Most of these suggestions pertain to issues of language.

Title:
1: Use “rhinoceroses” instead of “rhinos” and follow with “(Mammalia, Perissodactyla)”.

Abstract:
22: “present” instead of “propose”
28: “with a significant effect” instead of “to an important impact”

There are a number of other places in the text where “impact” is used and should be replaced with “effect.” Similarly, there are numerous places where the wrong preposition is used (e.g., “to” instead of “with” or “on” instead of “of”).

29: Delete “the” before “body mass.”
31: “greatest” instead of “maximal”
32: “of” instead of “on”
35: The phrase “reinforcement of the main lever arms” is unclear and probably not appropriate. I am guessing what is being described is having more pronounced attachments for muscles, or perhaps it is just the thickening of long bones to resist stresses produced by muscles. Either way, it is not clear from this phrasing.
37: “depending” instead of “pending;” this error occurs in other places throughout the text.

Introduction
46: “exhibit evolutionary convergence” instead of “present a…trend”
54: Delete “a” before “stylopodium” and “an” before “autopodium.” There are many places in the text where “a/an/the” should be deleted.
63: “extensively” instead of “massively”
76: “the peculiar morphology of hippos” instead of “hippo’s peculiar morphology.”
79: The word “consensual” is used several times in the text; replace it with “agreed upon.” Likewise, “non-consensual” can be replaced with “debated” or something similar.
84: “refined” instead of “sharpened”
86: “some” instead of “ones”
87: “support” instead of “sustain”
91: “maximum” instead of “max”
95: “an aquatic” instead of “a water”
103: “for understanding evolution towards large body mass” instead of “to understand evolution towards a high body mass.”
104: “some” instead of “ones”
105: “explore” instead of “explored”
108: “published” instead of “proposed”
109: “emphasized” instead of “aimed to emphasize”
110: What is meant by “determination criteria?”
111: “variation in” instead of “variations on”
114: “three dimensions” instead of “3D”

Materials and Methods
131: “included” instead of “kept”
143: “sex” instead of “gender”
156: Delete “the” before “Avizo”.
161: “come from” instead of “were borrowed to”
164: “specific” instead of “precise” and “complemented” instead of “completed”
169: “complemented” instead of “completed”
171: “that of hippos” instead of “hippo’s”
194: “procedures such as” instead of “procedure as”
209: What is “preconized”? Perhaps “conceived”?

Results
254: Insert “with” before “very” and “of” before “the diaphysis”.
265: Insert “and” before “a capitulum”.
370: “cluster’s” instead of “clusters”
384: This is one of several places where “expended” is used when “expanded” is meant.
389: “a” instead of “at”
393: “trapezoidal” instead of “trapezoid”
415-416: The sentence starting “D. bicornis” and ending “both axes” is unclear and needs to be fixed.
590: What is meant by “smooth details”?
602: Delete “the” before “group” and “allometry”.
603: Delete “the” before “species”.
604: Delete “the” before “centroid”.

Discussion
631: “allows for clear isolation of” instead of “enables to clearly isolate”
635: Delete “the” at end of line (before “phylogenetic”).
637: Delete “both”.
639: “depending” instead of “pending”
642: “debated” instead of “non-consensual”
674: Insert “the” before “shoulder”.
679: “on the other hand” instead of “at the opposite”
689: Delete period after “trochanters”.
690: “supporting” instead of “endorsing”
693-699: Mihlbachler and colleagues have studied knee locks in rhinos, but I think the only thing in print is an abstract (Shockey et al., Journal of Vertebrate Paleontology vol. 28, supplement to no. 3, p. 142A).
695: Delete “the” before “openness” and “habitat”.
700: “seems” instead of “seem”
703: “with” instead of “to”
704: “a” instead of “an” before “horizontal”
709: Delete “the”.
756 “Paired zeugopodial” instead of “Zeugopodial paired”
757, 760, 762, and 775: “expands” instead of “expends”
760: “caudal” instead of “caudally”
772: “Domning” instead of “Domming”; change this in References as well.
788: “appears greatest” instead of “seems maximal”
789: “On the other hand” instead of “At the opposite”
801: “proximal fibular epiphyss” instead of “one of the fibula one”
803: “fibular” instead of “fibula”
804: “covary” instead of “variate conjointly”
817: Delete “the” before “tibia”.
819: “perhaps” instead of “maybe” and delete “the” before “load”
827: What is meant by “tallest”? Largest centroid value?
841: “is” instead of “are the”
842: “Rhino” instead of “Rhino’s”
860: “ulna’s” instead of “ulna” and “extensively” instead of “extendedly”
870: “the hind” instead of “Hind”
880: “The rhino” instead of “Rhino’s”
881: “to that of the horse” instead of “to the horse’s one”
885: “address” instead of “state on”
887: “for” instead of “to” and insert “to” before “rise”
888: Delete “the” before “robustness”.
890: “with” instead of “to”
898: “to agree” instead of “coherent”
914: Delete “latter”. Replace “also” with “are”.
916: “variation” instead of “variations”
916-918: “whereas zeugopodial bones, especially the radius and ulna, are more strongly affected by body size, which indicates their important role in weight bearing.” instead of “whereas it is more…weight bearing.”
918: “The shape of the tibia is influenced by both body size and phylogeny.” instead of “As for the shape…phylogeny.”
920: “significant” instead of “the important”
921: “Comparisons” instead of “Quick comparisons”
923: “description of” instead of “to describe”
Experimental design: The methods are appropriate, as are the conclusions drawn from the results. My only substantial criticism regards how the authors distinguish “body size,” which appears to be represented by centroid size, from body mass, which is based on average species masses from the literature. I would argue that centroid size and body mass are both proxies for the more abstract concept of “body size.” I would therefore suggest replacing “body size” with “centroid size” whenever the reference is to the empirical observation of centroid size, and replacing references to “body size and body mass” with just “body size,” at least when centroid size and body mass have similar effects on variation.
Validity of the findings: The conclusions are appropriate, though they suggest a need for further comparisons with other large-bodied terrestrial mammals, especially since the authors preface the paper by discussing and emphasizing graviportal adaptations.
Additional comments: No additional comments