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: RESIDUAL DYNAMICS AND DIETARY EXPOSURE RISK OF DIMETHOATE AND ITS METABOLITE IN GREENHOUSE CELERY
Review round: 1
Reviewer: 1

Output:
Basic reporting: The language of the manuscript is good. The introduction is sufficiently detailed, However, there is a contradiction in it namely the statement on dimethoate residues seriously affected human health. This statement has no ground and contradicts the conclusion. It should be removed or modified
The manuscript is well structured, the figures and tables are technically appropriate, however the values reported are questionable.
The topic selected is actual and relevant, however the reporting the experiments and the analyses is not providing the necessary details to judge the reality of the results.
Detailed comments are given in the manuscript.
Experimental design: Dimethoate 40EC was used for treating the crops. At day 0 0.16-0.18 mg/kg omethoate was detected. It warrants to check the purity of 40EC formulation and report its omethoate content
Pesticide application: specify application equipment, its calibration, and report the actual dose delivered.
Pls specify the time interval required for celery to reach mature stage form seedling, planting and middle growth stage.
Check the registered use pattern of celery. Applying 20 times higher dosage than recommended in EU seems questionable.
Sampling: specify the portion of commodity sampled, report the number of primary sampling position from which the 2 kg laboratory sample was collected at each sampling time
How long the harvesting period lasts? What is the practical relevance of applying the pesticide 2-3 times at 7 days interval? What is the relevance of such application in view of the official preharvest interval?
What is the time between middle growth stage and reaching the maturity of celery?

Laboratory sampling operations: specify the part of the mature celery analyzed, the portion of the laboratory sample comminuted and the equipment used.
Analysis: what is the reproducibility of determination of the compounds? Recovery studies does not reveal any information on the reproducibility of analyses on which the decline studies were based.
Describe performance of recovery tests. At day 0 about 4 mg/kg after 3 days 1.7-2.0 mg/kg residues was measured??? The highest level of recovery test was 1 mg/kg.
The LOQ, LOD of the method should be reported.
There is no point to compare average of residues measured in three replicate samples. Residues in each sample should comply with MRL.
Exposure assessment. It is not clear what was the basis and relevance of estimating median residues for dietary risk assessment.
What was the reason to disregard median residue in equ. 5 considering the consumption patterns reported?
Validity of the findings: Based on the reported data the validity of the results and conclusions cannot be judged. See detailed comments above
Additional comments: In summary: based on the applied experimental design and analyses of samples reported, the validity of the study cannot be judged, and the credibility of the conclusions is rather questionable.
If the missing information is available for the authors and the experimental part of the study can be properly reported addressing the above missing points, then a major revision of the manuscript might be worthwhile. Otherwise the manuscript is not suitable 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: RESIDUAL DYNAMICS AND DIETARY EXPOSURE RISK OF DIMETHOATE AND ITS METABOLITE IN GREENHOUSE CELERY
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The study investigates a relevant issue and is well writen and professional.
Please see minor corrrections and suggestions regarding the conclusions to be emphasized in the atteched pdf document in the comments.
Congratulations and best regards.


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: RESIDUAL DYNAMICS AND DIETARY EXPOSURE RISK OF DIMETHOATE AND ITS METABOLITE IN GREENHOUSE CELERY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors tried to improve the description of experiments, but there are several major discrepancies which have to be sorted out before the paper can be accepted for publication.
Experimental design: The text does not provide clear information on the implementation of pesticide applications and sampling days
Validity of the findings: It cannot be judged before the experimental design is clarified and precisely described.
Additional comments: General comment: Please clearly link growing stage, days of pesticide applications, days between last pesticide treatment and sampling and residues measured. Perhaps a graphical presentation would help to make the experimental design and it description unambiguous.
Specific comments see also attached manuscript with tracked changes
L. 19-27 Since omethoate derives from dimethoate, there is no point in reporting residues after 28 days, when the PHI for dimethoate is 14 days. What is the justification for calculation of acute intake after 21 days and chronic intake after 28 days? Pls. state clearly what is the registered PHI for dimethoate in celery and perform the calculations for that (or shorter) interval.
L. 39. the problem here is not 550,000, but the use of “h” (hecto) instead of internationally accepted units. The description of application rate above is also confusing and should be corrected.
L.133. How many days after last pesticide application was the sampling performed? How pesticide applications were made before ’one time point sampling’?  Please clearly separate the trials with sequential sampling and ’one time point sampling.
L243-245: pls. revise the sentence: state MRL of dimethoate and compare the residues measured to the MRL. Dissipation rate has no relation to MRL.
L.258. Pls. reconsider this conclusion: If the Day 0 samples were taken shortly after the first application of dimethoate, the omethoate concentration should be practically close to LOQ, then gradually increase as the dimethoate degrades. So it is a complex time-conc. relationship and describing it as 1st order reaction is a very rough estimation. Probably from day 3 the decline could be described as 1st order.
L. 269 Reconsider Table 4. Clearly indicate the days of pesticide application and the growing stage of crop together with the residues measured
L. 273. This paragraph does not reflect the table. Both should be reconsidered and revised accordingly.
Table 5. Indicate the days of pesticide treatment in connection with sampling days
Table 6. The title of the table and it content do not match
Table 7 To consider total vegetable intake  (instead of celery) provides a very uncertain result because there is no information on the residue content of other vegetable consumed on the same day. This uncertainty shall be pointed out in the article
Discussion section cannot be evaluated until the elucidation of unclear points indicated above
L.368. perhaps: would lead to omethoate residues which would not risk the health of consumers
L. 373 It was indicated that the dissipation rate of dimethoate and omethoate residues is affected by several factors. What is the basis of such a general statement?


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: RESIDUAL DYNAMICS AND DIETARY EXPOSURE RISK OF DIMETHOATE AND ITS METABOLITE IN GREENHOUSE CELERY
Review round: 3
Reviewer: 1

Output:
Basic reporting: The write-up has been improved but still there are a number of points to be corrected. They are indicated in the annotated manuscript.
No attempt was made to edit the language.
Experimental design: It has been clarified and now the experiment results can be evaluated
Validity of the findings: The analytical part is marginally acceptable. However, the risk assessment would require full revision.
Additional comments: The word document with tracked changes do not correspond with the pdf document. The applied dosage is not clear. Is it 15 g formulated product per 100 m2 or 15 g active substance/100 m2
Abstracts and text hectare is an area (10,000 square meter) it should not be put on second power ha2
L. 80. specify dosage in g active ingredient (ai.) or formulation.
Repeatability of recovery tests does not reflect the uncertainty of reported residue values. To determine the uncertainty of reported residue values several test portions should be taken from blended material, and the relative standard deviation of the replicate measurements of residues in treated celery should be calculated and reported together with the measured residue values.

Risk assessment is incorrect as the sum of residues of dimethoate and X×omethoate should be taken into account (X is to account for the higher toxicity of omethoate). Note that the toxicological endpoints are the same so the parent compound and its metabolite should be considered together for both acute and chronic dietary intake (for instance see Tables 5 and 6)

Figure 3. It is not correct to describe omethoate dissipation as 1st order reaction including the first 3 days (at day 0 omethoate concentration should be below LOD). The explanation provided is not sufficient for supporting the calculation presented.
Table 4. sub-title is misleading: just write seedling stage and place a footnote: final residues following treatment at seedling stage; same comments for other sub-titles
Table 7: the calculation of STMR values should be explained in footnotes or in the text. As indicated above the sum of dimethoate and omethoate shall be considered.
Moreover, please note that MRLs are not safety limits and should not be used as reference point for evaluation of dietary risk.
Table 7 and 8: why the STMR values are different in the tables?


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: RESIDUAL DYNAMICS AND DIETARY EXPOSURE RISK OF DIMETHOATE AND ITS METABOLITE IN GREENHOUSE CELERY
Review round: 3
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The study investigates a relevant issue and is well written and professional.
Unfortunately, some more corrections are needed for tha manuscript to be accepted. Please see suggestions for minor corrections in the attached pdf document in the 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 REGULATORY ROLES AND MECHANISMS OF THE TRANSCRIPTION FACTOR FOXF2 IN HUMAN DISEASES
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. References are missing in some instances. For eg. Line 89-90 "FOXA1......identity.".

2. Although the introduction is mostly descriptive , the information listed at times is tangential and does not provide a direct connection. For eg., Line 96-99 " In the early..............is not expressed. Here authors have used words like major role and greater function rather than specifically mentioning the "role" and "function".

3. Line 129 Abbrevivated form 'EMT' is used without the description. Such an instance is repeated multiple times.

4. Line 131, The word "active" in that sentence does not seem right. Authors might want to use "activate" instead.

5. Line 159- 'FOXF2 was downregulated' should be presented in present tense. Such an instance is repeated multiple times and is almost present in the entire text.

6. Line 261 "The specific statement has been mentioned in the article of Lo (Lo 2017)." Which specific statement is author talking about. Authors should present their view on such findings rather talking about interpretations from a primary literature.

7. Line 280-281 "As mentioned above, FOXF2 is a tumor suppressor in cervical cancer." Not clear which part of the review is author refering to.

8. A schematic of FOXF2-mediated signalling in different cancers would be helpful for the readers.
Experimental design: Refernces are missing in some statements. Authors should carefully verify that refernces are appropriately placed.
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: THE REGULATORY ROLES AND MECHANISMS OF THE TRANSCRIPTION FACTOR FOXF2 IN HUMAN DISEASES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript “FOXF2: A key regulator in health and cancer” by Wu and You describes the role of FOXF2 in the development of various organs during embryonic development. The abnormal expression of FOXF2 results in dysregulation of various signaling pathways which further leads to the progression of various types of cancers. Authors have first described the importance of FOX transcriptional factor family and then detailed the effect of aberrant FOXF2 expression in relation to cancer in different organs. The mechanism of cancer progression in lungs, digestive tract, breast cancer, cervical and prostate cancer associated with FOXF2 expression has been explained. The pathways affected by FOXF2 and their effect on cancer progression are described in detail. The manuscript is concluded with author’s view to target FOXF2 for the treatment of different cancers. Overall, this review is very informative and provide the complete information of FOXF2 as a regulator in cancer. This is a well written manuscript.

Unfortunately, the study is not new. A similar review has been published on the role of FOXF2 by He et al. in the journal “Cell Death and Disease” on June 5, 2020. They have described each and every aspect of FOXF2 in embryonic development as well as in various cancers. The signaling pathways through which FOXF2 might act, have also been explained very well. The current manuscript looks very similar to this one. How do authors explain this? What is new in current manuscript in addition to this above-mentioned review article? Authors should revise the manuscript so that it does not look just a repetition of already published review.

Authors should make at least 1 schematic figure to show the role on FOXF2 in various cancers and the signaling pathways involved in which it can be indicated where the upregulation or downregulation of FOXF2 affect a particular pathway that leads to a specific type of cancer.

Manuscript should be checked thoroughly for English word choice, grammar and sentence organization. Several sentences seem to be repetitive throughout the manuscript. Also, at multiple places, the text seems discontinuous.

Authors have mentioned that FOXF2 can be a good target for the treatment of cancer, though the strategy to do so is not described. FOXF2 regulation is highly important, so its reduced or enhanced expression will not be a cure for cancer. What do authors think about a possible mechanism which can be used to target FOXF2 for cancer therapeutics?
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: THE REGULATORY ROLES AND MECHANISMS OF THE TRANSCRIPTION FACTOR FOXF2 IN HUMAN DISEASES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised submission of the manuscript entitled “The role and regulatory mechanism of FOXF2 in diseases”, features the regulatory mechanism of FOXF2 in cancer as well as non-cancerous diseases. Authors have answered all the comments/concerns point-wise and improved the manuscript accordingly. In the revised version of the manuscript, authors have added three schematic figures for better understanding of the mechanism.

English language still needs improvement. At multiple places, it is hard to understand the sentences.

Authors have used the word “non-cancer diseases” multiple times in the text. It should be rephrased.

Title should be modified to be more specific. Disease is a very broad term. Please do specify the title.

Line 44: “It has been demonstrated that abnormal transcription factor regulation affects the expression modification of their regulatory genes, which is a mechanism required for the formation of human malignant tumors.” This line is not clear. Please do modify it.

Figure 1 and 2 look good. However, figure 3 is bit confusing. How SUFU inhibits GLI outside nucleus and afterwards figure displays GLI and FOXF2 in nucleus which is not clear. Please do modify the figure accordingly so that it describes everything clearly.
Experimental design: no comments
Validity of the findings: no comments
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 REGULATORY ROLES AND MECHANISMS OF THE TRANSCRIPTION FACTOR FOXF2 IN HUMAN DISEASES
Review round: 3
Reviewer: 1

Output:
Basic reporting: N/A
Experimental design: N/A
Validity of the findings: N/A
Additional comments: Congrats


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 REGULATORY ROLES AND MECHANISMS OF THE TRANSCRIPTION FACTOR FOXF2 IN HUMAN DISEASES
Review round: 3
Reviewer: 2

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comments
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: UNDERCARBOXYLATED OSTEOCALCIN INHIBITS THE EARLY DIFFERENTIATION OF OSTEOCLAST MEDIATED BY GPRC6A
Review round: 1
Reviewer: 1

Output:
Basic reporting: This article is clearly written and includes all the relevant papers are cited
Experimental design: This manuscript presents some level of novelty. Scientific question is well defined and methodology is described.
Validity of the findings: If the recombinant osteocalcin used in this study is sufficient, conclusions are supported with the results. Statistic is adequate.
Additional comments: This manuscript investigates the effect uncarboxylated osteocalcin on RAW264.7-derived osteoclasts. Overall, this study is well conducted and the experiments are correctly designed Also this study adds some novel aspects which were not investigated before. However, one major concern, which should be addressed before the publication is the purity of the osteocalcin used in this study. The authors generated recombinant osteocalcin using bacterial expression system and measured its concentration using specific ELISA. To draw the conclusion that the effects observed in this study are related to osteocalcin the authors need to prove that their recombinant protein do not contain contaminants of other proteins or components of the bacteria. They should perform an SDS page with purified osteocalcin followed by coomassie blue staining. They should also test the solution with recombinant osteocalcin for any bacterial components.


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: UNDERCARBOXYLATED OSTEOCALCIN INHIBITS THE EARLY DIFFERENTIATION OF OSTEOCLAST MEDIATED BY GPRC6A
Review round: 1
Reviewer: 2

Output:
Basic reporting: I found the text poorly written and difficult to follow. Major revisions of the written English are required.
Figure 1A is difficult to interpret. For me is much more intuitive to plot a time vs OD curve, distinguishing the different concentrations with different curves.
Line 61 to 66. This sentence is completely incomprehensible to me. Why should OCN have a role, either carboxylated or not, in OCN-deficient bone particles (BP) resorption? What are bone particles? Where are they found? What is their pathophysiological significance?
Experimental design: The overall experimental session lack of a comparison between carboxylated OCN and ucOCN, being mainly focused on the latter. This is a key point to understand the physiological role of the inhibitory effect of ucOCN on Ocl. In other words, why should Ocl be inhibited by the product of their activity? (considering that, at least in mouse, bone resorption is partially stimulated by osteoblasts through the action if insulin).
Validity of the findings: The choice of 100 ng/mL of ucOCN to investigate the mediation of GPRC6A is questionable since the average serum concentration of ucOCN is about 10 ng/mL. Are there any studies reporting bone concentration of ucOCN greater than the serum?
Additional comments: In this paper, Wang et al. investigate the possible inhibitory activity of undercarboxylated osteocalcin (ucOCN) on osteoclast (Ocl)differentiation and function. Authors find that ucOCN inhibits Ocl precursors proliferation, migration and differentiation into mature Ocl. In addition, ucOCN seems to inhibit the bone remodeling function of this cell population. All these effects are mediated by GPRC6A.
It is a quite interesting study, disclosing some novel effects of osteocalcin in its main tissue of expression. However, I have many concerns that require to be 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: UNDERCARBOXYLATED OSTEOCALCIN INHIBITS THE EARLY DIFFERENTIATION OF OSTEOCLAST MEDIATED BY GPRC6A
Review round: 2
Reviewer: 1

Output:
Basic reporting: This article is clearly written and includes all the relevant papers are cited
Experimental design: This manuscript presents some level of novelty. The scientific question is well defined and methodology is described.
Validity of the findings: Conclusions are supported by the results. The statistic is adequate.
Additional comments: All of my concerns have been 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: PUTATIVE CARBOXYLESTERASE GENE IDENTIFICATION AND THEIR EXPRESSION PATTERNS IN HYPHANTRIA CUNEA (DRURY)
Review round: 1
Reviewer: 1

Output:
Basic reporting: o	In terms of English quality it looks regular. Some words seem odd in this type of research field, i.e. “olfactorily” in the abstract (Line 25). Similarly, in Line 31 “our results indicated” should be substituted by “our results indicate”. Line 245 “Till now” is not proper. There are some typos: see line 111 (“ware”). I hence would recommend to send the manuscript for English revision to a native speaker researcher from our line of study.
o	In line 62, I do not consider that the part of “as well as herbicide activation” is pertinent because it seems out of place in this kind of manuscript. Furthermore the citations on this part refer much more to insecticide resistance, which although there is a body of publications about herbicides, could lead the reader to mistrust if they meant herbicide or insecticide.
o	Line 69, should include, of course, Drosophila melanogaster, they can use the reference of Chertemps 2012 or the one from 2015.
o	At line 80 they should add as well the main findings on Chertemps et al., 2015.
o	Line 95, I think is pertinent to write a couple of lines about the identity of the pheromone components of the pheromone blend of H. cunea.
o	At line 97, the citation is not italics as all the rest.
o	Re-write the sentences between 125-128 because as it is right now don’t make any sense.
o	Line 196, leave it until “subclass” the rest is rather obvious.
o	Figure 1 legend needs major corrections: first try to state under the figure the type of tree (Maximum likelihood), the model used (see my comment on substitution model) and the bootstrap, do not only say to go back to the methodology for details. Indicate that the numbers are also placed on the tree to indicate bootstrap support.
o	In the phylogeny analysis part in results it would be useful to state the percentages of identity between HcunCXEs and Dmel, Sinf and Slit at least for the most closely related.
o	In line 221, it must be acknowledged that the sex-biased expression is not statistically significant but there is a clear numerical difference between expression in the sexes.
o	Lines 235-239 are not really relevant in here. If they want to introduce the importance of ODEs they should use the explanation given by Leal (2013) on how ODEs must act fast for plume determination.
o	Lines 240-241: how much homology? What percentage of identity? This statement is not obvious just by looking at the tree.
o	Line 244: this is a good time to mention the pheromone components.
o	Line 262: but it could also act in a broader function. They should see paper from 2015 and related to their results.
o	Line 287: correct this scientific name! the species name is ALWAYS in lowercase letters
o	It is important that they share the protein sequences as raw data. This is also important for reviewers.
Experimental design: I think it does comply with the aims and scope, the research question is well-defined and meaningful. However, I have some problems with the methodology used and described:

1.My main question is how did they find the 10 ODEs? They mention that they use the information from their 2016 transcriptome, but I want to know how they performed the extraction of these candidate genes. As it is right now, they just state that used the transcriptome to check for transcript abundance but it does not make clear how they actually annotated candidate ODEs. I guess they just ran some kind of BLAST to their previously determined transcriptome, but it is important to state it, along with the parameters to consider as reliable predictions (e-value, % identity, hits, etc). I would add a whole new section “Gene annotation” or something similar. Furthermore, I want to know what method did they use to name the new genes? Based on what? It does not look like they based it on transcript abundance or similarity to other species sequences (S. inferens for example), so please explain.
2. I have a problem with the methodology followed to construct the phylogenetic tree. The first thing I recommend is to go ahead and state in lines 131 and 132 all the species from which ODE sequences were used to make the comparison. I also personally prefer MAFFT for multiple sequences alignment but I guess Clustal is fine. It is however necessary that they perform a best substitution model analysis to make the phylogenetic tree, after all, choosing the proper evolutionary model can affect the topology of the tree, branch length estimation the statistical support values. They also will need to state in the figure or elsewhere which is the selected gene evolution model. Considering that there are several newer versions of MEGA I would also recommend to use newer versions if possible.
3. I would recommend to state how they normalize the data of relative RNA expression in the methodology.
Validity of the findings: No comment
Additional comments: Your article fills up an important gap since ODEs have not been as extensively investigated as other chemoreceptor proteins for example. Nevertheless, I strongly recommend to implement a series of small changes to make it more scientifically sound. First and most importantly to check again for language mistakes and some others changes to increase the reliability of your findings.


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: PUTATIVE CARBOXYLESTERASE GENE IDENTIFICATION AND THEIR EXPRESSION PATTERNS IN HYPHANTRIA CUNEA (DRURY)
Review round: 1
Reviewer: 2

Output:
Basic reporting: a.	Language: There is a high number of spelling/grammatical errors throughout the entire manuscript including Tables and Figures. There are also some typographical errors such as extra spaces. (eg. Line 102) Several statements must be improved for clarity and overall understanding of the text (Please refer to the highlighted text in manuscript pdf and comments below). The following lines represent examples for which language could be improved:
Line 53-54: This statement can be improved for clarity and also requires a reference.
Line 55-59: This statement can be improved for clarity and scientific accuracy. These families of enzymes are distinctly different based on their protein structure and function; however, they have been shown to catalytically interact with odor molecules of the same type and structure; ie.  it is currently believed that multiple ODEs (belonging to the different enzyme families) may participate in degradation and clearing of the same type of odor molecule (See Steiner et al. 2019)
Line 65-67: This statement can be improved for clarity and grammatical correctness.
Line 100-102: This statement can be improved for clarity.  Is it the differential expression pattern that suggests the function?  Is the expression of the gene enough to suggest a function?
Line 185:  It is not clear why the CXE genes were specifically compared to S. inferens.
Line 217-218: RT-qPCR can’t be used to probe the physiological function of HcunCXEs, it can be used to explore differential expression patterns of the transcripts of interests between different tissue types and sexes.  The statement should be reworked for clarity of what is being investigated.  Inferring physiological function from transcript expression level is speculative at best and must be clearly identified as such.
Line 251-254:  The OR landscape of H. cunea is known from the 2016 transcriptomic analysis and should, therefore, be referenced here.  This statement should also be clarified for the readers.  The number of CXEs identified is not that surprising given our current understanding of CXE’s function in clearing volatile molecules from the sensillar lymph.  Please refer to Steiner et al 2019, Durand 2010a,b Durand 2016.
Line 282-294: How are the H.cunea results relevant or related to the content of the discussion here?  The authors could improve this paragraph by providing relevant explanation to the results from H. cunea.
b.	References: Several statements are missing relevant references. (Example: Line 53-53; Line 64-65) vi.	The references list lacks in consistency and accuracy of the information provided. Some of the journal titles are in italics and some are not, some are represented as abbreviations and some are not.  The titles are also not consistently capitalized.  The references also contain a number of spelling and typographical errors.
c.	Professional article structure, figures, tables:  The structure of the paper is well organized and logical. The figures contain spelling errors and several of the figures could be represented better. Raw data shared is adequate.
Fig 1: The figure could be improved by having the Neurosignalling (red) and Generally intracellular enzymes (green) aligned parallel with the page and the two labels in blue.  The world generally can likely be omitted for clarity.  The figure description should be able to stand alone without having to refer to materials and methods.
Fig 2: The figure could be improved by introducing a and b for the distinct portions of the figure to avoid an ambiguous statement such as “the upper parts listed.” The description which follows could be reworked for better clarity.  It is not clear what 8 most common motif-patterns are. The word Homologous in the figure is spelled incorrectly.
Fig 3: The y-axis in this figure for all of the graphs shown contains a spelling error.  The clarity of the figure could be improved by combining results for all CXEs for one tissue type on one graph to immediately be able to know which of the genes are abundant in the given tissue type.
Fig 4:  This figure would benefit from having the tissues involved in RT-qPCR included for comparison purposes.  This would provide a link for the two experiments and further shed light on the relative expression of the CXE genes in the different tissue types.
Tables 1 & 2: the descriptions should include an explanation for any abbreviations used in the table so that the tables can stand alone without having to refer to the text of the manuscript.
d.     The descriptive work with H.cunea CXE sequences is relevant to the questions regarding signal deactivation in odorant processing.
Experimental design: a.	The research presented falls within the aims and scope of the journal but many issues should be addressed before the paper can be published.
b.	The research question is well defined, relevant & meaningful
c.	No comment
d.	Methods are not described with sufficient detail & information to replicate the experiments.
Line 107-109: The description of insect material collection, rearing, and tissue collection should be improved for clarity.  It is not clear what the authors mean by 1st generation population and if the pupae collected at the site listed were reared to adulthood.  Were the tissues collected from the 1st generation adults or several generations?  There is no mention of diet or reference to previously established rearing protocol. Were the pupae and larvae used for tissue collections from the materials collected at the Baimao Town site or as offspring from the 1st generation adults?
Line 116-119:  The software package version as well as parameters used in RSEM transcript abundance quantification are missing from the description for future reproducibility.
Line 123-125: Software version, reference, or web address as well as relevant parameters such as the minimum transcript length specified for the ORF prediction should be listed.
Line 133-134:  MEGA5.0 parameters for the neighbor-joining method should be provided.  What program was used to generate the final graphical tree representation and was the final edited?
Line 109-111 and 145-149: How many insects (ie. number of males and females) were the tissues extracted from? The authors could also describe the excision points for each tissue type for clarity and reader understanding, ie. what was the excision point for the antennal tissue? The TRIzol extraction protocol was performed according to the manufacturer’s protocol or was it modified in any way? It would be helpful if the overall description for the RNA extraction and cDNA synthesis provided more detail as there are several ways the purified RNA can be treated before cDNA synthesis.
Line 154-156: It would be helpful if authors could specify whether whole pupae and larvae were used in tissue collection as well as the number of insects used.
Validity of the findings: a.	No comment.
b.	The authors state that this study is performed based on previous trancriptomic work from 2016.  Search in NCBI databases reveals two transcriptomes PRJNA513855 and PRJNA428142.  The authors of this study should specify which transcriptome was used in this study using the proper accession number rather than SUB6944247.  The raw data provided is adequate.
c.	The results and discussion sections could be improved by emphasizing how the results from H. cunea correspond to previous findings in other insect species.  Several statements lack precision and clarity (Please refer to Section 1 for examples) It would be helpful if the authors could reiterate in the conclusion which of the 10 CXEs had antennal expression and based on the results taken together were most likely to be implicated in signal deactivation at the level of antenna.
d.	The authors do make several speculative statements that require clarification and identification as speculative.
Additional comments: 1.	The importance/relevance of conserved motifs in CXE protein sequences is not mentioned in the introduction.  Thus, when it is mentioned in materials and methods as well as results it is difficult for the reader to understand why this analysis is being done in the first place. Perhaps a brief mention of the CXE motif structure characteristic to the different protein subclasses might help clarify the usefulness of this information.
2.	The authors mention RSEM transcript abundance estimation in the materials and methods however, the results from this analysis are not used in the results and discussion section.  If this analysis is not relevant it should be removed from the Materials and Methods section for clarity. I note that the FPKM values are reported in Table 1 but these results are not addressed in the text of the results and discussion. How do these results compare to quantitative PCR? How are they relevant to the analysis overall?
3.	Consistency in writing: in some instances, the authors provide the version of the program or the manufacturer of the reagent but in many cases, they do not.


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: PUTATIVE CARBOXYLESTERASE GENE IDENTIFICATION AND THEIR EXPRESSION PATTERNS IN HYPHANTRIA CUNEA (DRURY)
Review round: 1
Reviewer: 3

Output:
Basic reporting: There is a lack of clarity of the reporting of the materials and methods that preclude full analysis of some of the data being presented. This must be rectified before the manuscript can be adequately reviewed and assessed.

In general the language of the text is satisfactory. However, there are sufficient many grammatical errors that I recommend the authors ensure the manuscript is edited for language prior to re-submission.

Furthermore there are several minor errors in the presentation of the text and figures that indicate that the authors need to take greater care in attention to detail and these errors must be corrected before re-submitting. Specific examples are given below in the general comments to the authors.
Experimental design: This research study fits within the aims and scope of the journal.

Methods are not described with sufficient detail and information to replicate. Critical information is missing. Specific comments are provided in the general comments for the authors section below.
Validity of the findings: Underlying data has been provided, however, it is not clear in the supplemental data what the underlying data represents. Clarification is required. Specific comments presented below in the general comments to the authors.

It is difficult to assess the validity of the RT-qPCR data because MIQE guidelines for reporting aPCR studies have not been adhered to. Critical information is missing in the materials and methods that render it difficult to accurately assess the data.
Additional comments: Abstract.

Line 29. “previous H. cunea transcriptomic data”

The authors should clearly indicate here the source of the data. Which tissue. Later on in the article, it is mentioned that this is antennal transcriptome, but that should be made clear to the reader in the abstract.

Introduction

Line 53-54. “which may lead to disorders of the nervous system”.

This is vague and not clear what the authors mean. Greater specificity is required. For example, it could be said “which may lead to poor spatio-temporal resolution of the odor signal, and pose fatal hazards to the insects”

Line 66. Change “so as to” to “and”.

Line 97. “transcriptomic data”.

Again here, indicate that this is data from antennal transcriptome.

Line 99. “reverse transcription-quantitative PCR”.

Typically RT-qPCR refers to Real-Time quantitative PCR, not reverse transcription. Also abbreviations should be indicated in parenthesis foer both this and reverse transcription PCR, which follows.

Materials and Methods

Line 108. “Several body parts/tissues…”

It is essential to report how many insects were used for each tissue type. Also, how many biological samples were collected. This, especially, is important to know, with reference to the RT-qPCR study.

Line 131. “other reported insects were subjected”.

Which other insects. It is necessary in the Materials and Methods section to clarify which insects and motivate the choice of species, and specify where the sequences were obtained from (for example, from NCBI, or from directly from specific publications).

Line 131-132. “were subjected to multi-sequence alignment on Clustal…”
Line 133. “constructed using MEGA5.0…”

For Clustal, Mega, and all other bioinformatics software, it is necessary to indicated exactly which parameters were set before running the program to facilitate repeatability within the scientific community.

Line 138-139. “A total of 44 CXEs from…were used for identification”

Why were CXEs from these species chosen? Walker et al., 2019, BMC Genomics identified a larger repertoire of antennal expressed CXE genes in Spodoptera littoralis, for example. Using that as a reference set may provide more robust data.

Line 153. RT-qPCR and RT-PCR analysis section

How the RT-qPCR study, how many biological replicates were performed. From line 164-165, it is indicated that each reaction was replicated three times, but it is not clear if this is referring to technical replicates, or biological replicates. For this kind of study it is important to be clear in reporting.

Additionally, the authors need to consider MIQE guidelines for reporting on RT-qPCR studies. Bustin et al., 2009, Clinical Chemistry. The qPCR primers are mentioned, but there is no data shown on primer efficiency. In the materials and methods there is no mention of any control genes that have been used in the qPCR study. Control genes are essential. In the figure legend EF-1a is indicated, but in the supplemental data on the primers, both EF-1a and GAPDH primers are indicated. Was one of them used? Or both? According to MIQE standards, it is preferable to use more than one reference gene, but if only one is used, it must be shown that the reference gene shows stable expression across all tissue types.

Results

“shared a more recent common ancestor with the CXEs in D. melanogaster, S. inferens and S. litura”
Looking at Figure 1, it is not clear what basis the authors claim that the HcunCXEs shared more recent common ancestor with genes from D. melanogaster. The claim could surely be made for S. inferens and S. litura, but for every instance where homology is inferred for a D. melanogaster gene, the Hcun homologue is more closely related to genes from S. inferens and/or S. litura.

It also appears there is a taxa specific expansion of Hcun CXEs in the intra-cellular family. The authors should mention this more clearly in the results and discuss the implications of such expansions in the Discussion Section.

Line 210. “6-5-3-3-1-8-2-7-4”

Looking at Figure two, this looks like a typo. It seems there should only be one “3” here.

Discussion

Line 251-252. “Surprisingly, the number of CXE genes (n=10)…was much lower than those of other reported lepidopteran species:…”

Perhaps the authors should specify here if any of these other species utilize TypeII pheromones. Also should comment on why they think they may have found less ODEs….because of biological/ecological reasons or because of methodological reasons?
In looking at the Zhang et al., 2016 transcriptomic article on which this study is based, the numbers of ORs, GRs and IRs is indicative that the antennal transcriptome is fairly complete. However, to provide greater confidence in the completeness of the transcriptome, and thereby suggesting most of the CXEs in the transcriptome have been identified, I recommend the authors run BUSCO analysis on their transcriptome. gVolante server is a good source for this kind of analysis. https://gvolante.riken.jp/index.html

Line 291. “S. litur”

Should be “litura”?

Figure Legends and Figures

Line 518. “(Hyph)”

What is “Hyph”? Should be “Hcun”?

Figure 2. “Homolohous CXEs”

Should be Homologous. Also some of the Numbers of CXEs with each pattern is wrong. In the numbers column in the lower half of the figure. For the first row. (14) is indicated, but on the “Homologous CXEs column, it appears there is only 11. Likewise for the fourth row (16) is indicated, but it appears there is 15.

Supplementary Material 1 Raw Data
In the excel sheet it is not clear what the raw data numbers represent. This should be made clearer. What are those values indicating? Also the worksheet tab name is written in a Chinese script. For publication in an international journal, it should be written in English.

Supplementary Material 2 Raw Data

For the RT-PCR original gels, the columns need to be labelled so that the reader can understand what they are looking at. Which tissues were used as source of cDNA for amplification. This needs to be clear with labels on each column.


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: PUTATIVE CARBOXYLESTERASE GENE IDENTIFICATION AND THEIR EXPRESSION PATTERNS IN HYPHANTRIA CUNEA (DRURY)
Review round: 2
Reviewer: 1

Output:
Basic reporting: Looks much better now. However, there are pending revisions to be made, typos and statements to be changed for clarity. These are the ones I noticed:

Introduction
Please change "The molecule-bound odorant receptors (ORs) then convert the chemical signals into electrical signal that is... " by "The molecule-bound odorant receptors (ORs) then convert the chemical signals into electrical signals that are"

Please notice this sentence is twice right now: We found that HcunCXEs displayed a either antennae-  or leg/wing-biased expression. We found that HcunCXEs displayed either antennae- or leg/wing-biased expression.

M&M
The sentence: "Genes related to the ODEs CXEs of H. cunea and other reported insects of Seasamia..." must be changed to "Genes related to the ODEs CXEs of H. cunea and other reported insects (Seasamia.....)"

Please indicate after "Mafft" that the most suitable evolutionary model was calculated with "X" program and then the phylogenetic tree constructed

Discussion
I do not understand this sentence: “HcunCXE2, 3, 4, 5, 6, 8 and 10 were homologous to this (e.g. DmelCG10175) in D. melanogaster”

Change “In the current study, nine out of 10 HcunCXEs were found to express in the larvae (Fig. 4)” for “In the current study, nine out of 10 HcunCXEs were expressed in the larvae (Fig. 4)”

Figure legends
Genrally secreted enzymes, substrates include hormone and pheromones; B: Generally intracellular enzymes, dietary metabolism/ detoxification functions; C: JHE; D: Genrally secreted enzymes, substrates includes hormone and pheromones; E: Nerouligins; F: ACHE.------------------------------correct “Generally”
Experimental design: Ok
Validity of the findings: Ok
Additional comments: Ok


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: PUTATIVE CARBOXYLESTERASE GENE IDENTIFICATION AND THEIR EXPRESSION PATTERNS IN HYPHANTRIA CUNEA (DRURY)
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors sufficiently addressed previous comments on the language.  The readability of the paper had improved greatly. I have suggested a small number of a single word or sentence structure changes in the attached pdf which may improve the overall clarity and readability of the paper however, these changes are rather minor.

Figure 1.
For clarity the bootstrap values should be represented as percentage or fraction rather than whole integers. For example see PNAS November 12, 1996 93 (23) 13429; https://doi.org/10.1073/pnas.93.23.13429
Is there significance associated with the thickness of the lines for the phylogenetic tree? If yes it should be addressed in the figure description, if not all lines should have the same thickness.
The bootstrap reference should be removed from the figure description or modified.
Experimental design: The manuscript meets all the criteria set out by PeerJ Review.  The authors have addressed all issues listed in the previous review.
Validity of the findings: The study is valuable and provides interesting results on the antennal CXEs in H. cunea.
The study provides interesting insights for both the species under investigation as well as the basis for future comparative studies for degradation of Type 1 and Type 2 pheromones.

The conclusions based on the gene expression analysis can be further improved for clarity.
The authors need to combine their results from phylogenetic analysis and expression studies to clearly delineate the candidates for pheromone/volatile degradation and candidates for basic metabolic functions. Please see General Comments section for details.
Additional comments: Overall, this is a very nice paper and a valuable foundation for future work on CXEs.  The highlighted portions of the manuscript pdf have associated comments.

I would recommend the following changes to improve the content of the discussion section of the manuscript.

1. Phylogenetic analysis
For A. polyphemus and M. brassica antennal CXE a function as pheromone degradation enzyme has been previously established. If you included the sequences corresponding to these CXEs in your phylogenetic analysis it would be clear that CXE1 and CXE9 are obvious candidates for pheromone/plant volatile degradation.  This combined with your gene expression analysis can provide further insights into roles for each of these proteins based on their sex-specific expression patterns.

2. Gene expression analysis
Line 256: You state that "although the sex-biased expression is not
statistically significant, there is a clear numerical difference between expression level in the sexes." However, Figure 2 shows the statistical difference between mean values for CXE1, 3 and 10 and not CXE 9.  It is worth to note that the most abundant CXE genes in the antenna are NOT the extracellular  CXE  that likely participate in the volatile odorant degradation. The most abundant CXE's are likely involved in primary metabolic activities and it would thus make sense their expression is much higher than for the other specialized CXEs in the antenna. Furthermore, some of the CXEs in the metabolic class such as CXE are also highly expressed in other tissues. This is related to your comments in the line 322 where you state that the most highly expressed CXE coding sequences are likely involved in pheromone degradation which is misleading.
Line 273: 1). "We speculated that these H. cunea CXE genes mainly degrade sex pheromone components and host plant volatiles." This is inconsistent with your phylogenetic findings. See note in edited pdf.  Only 3 out of the 10 CXE are likely candidates for pheromone degradation. The statement should be revised to reflect that only some of the CXE may putatively participate in pheromone degradation.
Line 278: It is not clear what the authors mean by this statement. It has been previously shown that a single CXE can degrade a number of different pheromones/volatiles and the number of CXEs vary among different insect species.
Line 316-317: It would be beneficial to specifically state here which of the CXEs are antenna specific.
Line 321-325: Need clarification or rewriting.


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: ONTOGENY OF HIGHLY VARIABLE CERATITID AMMONOIDS FROM THE ANISIAN (MIDDLE TRIASSIC)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English is very good. I pointed out some minor problems in the pdf.
I suggested one additional paper to cite.
The structure is clear, methods well-explained and state-of-the-art.
The content is well intelligible.
Interpretations are backed up by data.
Experimental design: -original, i.e. this was not carried out in the same way before with ammonoid data
-research question is well-defined
-thoroughly gathered data
-methods well described in great detail
Validity of the findings: The ms does address an interesting topic. It has been suspected that diversity is artificially inflated in the fossil record (ammonoids and other groups). This hypothesis was tested and falsified for a Triassic ammonoid family here.
From a method and statistical point of view, I have no complaints.
Additional comments: The main thing you could improve is writing out the taxon names in figures 7,8, 10 and 11. There is always enough space and the reader immediately knows where the data is from.


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: ONTOGENY OF HIGHLY VARIABLE CERATITID AMMONOIDS FROM THE ANISIAN (MIDDLE TRIASSIC)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is scientifically written and clear and unambiguous, professional English is used. Most appropriate literature is cited with some rare exceptions (see annotated PDF).
Experimental design: The methods are appropriate – I would like to see a larger discussion/comparison on the use of weighted or unweighted approaches (this could be done by producing a table). Also as far as I understood you did pairwise comparisons of trajectories in developmental morphospace but I guess this could also be supplemented by doing pairwise comparisons of variance of individual species within the ontogenetic "trajectory" morphospace. Also the names are a bit confusing as developmental more or less is a synonym of ontogenetic (maybe use ontogenetic trajectory space versus developmental space).
Validity of the findings: The interpretations are appropriate. The underlying data and code are provided and robust. Their name should however be corrected as they are currently addressed as "Field study approval documentation" (already in use for the actual field study approval documentation).
Additional comments: I greatly enjoyed reading your paper and seeing such novel analyses. In additional to those remarks, i have 3 additional points
1)	Some relevant references are missing (see annotated pdf)
2)	It would be good to differentiate in most graphs the landmarks and semi-landmarks (e.g., Fig. 9-10) for clarity and completeness sake.
3)	A more philosophical point: are continuous differences in ontogenetic development sufficient to separate species? This is rather an interpretation which might be appropriate but not necessarily true. Who is to say that time-averaged assemblages belonging to the same species within beds which likely did not overlap during their lifetimes where not exposed to different environmental conditions during their ontogenies and therefore look morphologically different/had differences in ontogenetic development.  I would therefore suggest to express yourself therefore slightly more carefully that “traditionally defined species also differ in ontogenetic trajectories which might support the idea that might be different species”.

These and additional suggestions can be found in the annotated pdf.


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF ADP-RIBOSYLATION FACTORS ASSOCIATED WITH BIOTIC AND ABIOTIC STRESS IN WHEAT (TRITICUM AESTIVUM L.)
Review round: 1
Reviewer: 1

Output:
Basic reporting: English language and style are fine/minor spell check required

It is necessary to improve the resolution of the Figure 2 (A; B and  C) and the Figure 5A
Experimental design: No comment
Validity of the findings: no comment
Additional comments: ADP-ribosylation factors (ARFs) belong to Ras superfamily of small GTP-binding proteins (GTPases), which was reported to function in diverse physiological and molecular activities. These low molecular weight (21–24 kDa) proteins was classified into ARFs and ARF-like (ARL) proteins based on their functional characteristics and sequence homology.  ARFs are highly conserved proteins (>60% sequence identity), while ARLs are highly divergent (40–60% identity). ARFs was identified in several plant specie (Arabidopsis, rice, tomato, potato, maize, carrot, wheat, tobacco and barley), in which they play a relevant role in a broad-spectrum biological process, including growth and development, senescence and senescence-dependent recycling, tolerance to biotic and abiotic stresses.In the present manuscript, the role of ARF genes was explorated in development of wheat (Triticum aestivum L.). The wheat is one of the most important crops for human, provding staple food globally.
The authors conduced a systematic phylogenetic analysis of TaARFs and analyzed their expression profiles at different development stages and under different biotic and abiotic stresses.74 wheat ARF genes were identified and clustered in 7 sub-groups. TaRFA1 subgroup genes are strongly conserved and induced in response to biotic and abiotic stresses. TaRFA1 proteins seems to interact with protein phosphatase 2 (PPC2), a key protein in the abscisic acid (ABA) signaling patway. This evidence would seem to support the hypothesis that TaRFA1 plays a role in the wheat stress response.In my opinion, the manuscript is interesting and provides comprehensive insights into the structure, organization, evolution, and expression profiles of the TaARF gene family in wheat.
The title is clear and informative about the research carried out. The abstract includes all the main findings reported in the manuscript.The introduction provides sufficient background and include all relevant references. Experimental approach is correct and methods are adequately described. The results are clearly presented, only the resolution of the Figures 2A; 2B and 2C) and the Figure 5A should be improved. The discussion is supported by the results.
English language and style are fine/minor spell check required. Based on my assessment, I am in favour of the publication of the manuscript, whether the authors will provide an accurate revision of the English language and make clear the Figures 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: GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF ADP-RIBOSYLATION FACTORS ASSOCIATED WITH BIOTIC AND ABIOTIC STRESS IN WHEAT (TRITICUM AESTIVUM L.)
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript by Li et al is well written, clear and sufficient background and introduction provided. The manuscript is well structured, the data are solid and raw data provided.
Experimental design: The manuscript is focused on analysis of ADP-ribosylation factors in wheat, one of the most important crops, and provide data and tools for research community to improve crop tolerence to various stresses. The authors made throughout phylogenetic analysis of TaARFs as well as gene structure, location and expression analysis of TaARFs.
Methods are described with sufficient details and information. The results are clearly presented in figures.
One major concern about the experimental design is qRT-PCR data obtained under cold, salt, and ABA stress. According to the MIQE guidelines the use of at least two reference genes is nessesary for a strong and reliable normalisation in RT-qPCR experiments. The authors, however, use only one reference gene in the study. So, it should be performed according to the MIQE standards.
Validity of the findings: no comments
Additional comments: The only weakness of the paper is qRT-PCR data as indicated above, which should be improved before acceptance of the paper.


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF ADP-RIBOSYLATION FACTORS ASSOCIATED WITH BIOTIC AND ABIOTIC STRESS IN WHEAT (TRITICUM AESTIVUM L.)
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors modified the manuscript according to the requests of my first review


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

Input:
Title: GENOME-WIDE IDENTIFICATION AND EXPRESSION ANALYSIS OF ADP-RIBOSYLATION FACTORS ASSOCIATED WITH BIOTIC AND ABIOTIC STRESS IN WHEAT (TRITICUM AESTIVUM L.)
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: The authors have improved their manuscript by adding one more reference gene for qRT-PCR data normalization. As it was a major concern during first review, I recommend now publication of the paper.
Validity of the findings: no comment
Additional comments: The required additional experiments were done as suggested, the manuscipt can be now 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: TROPHIC NICHE DIFFERENTIATION AND UTILISATION OF FOOD RESOURCES IN COLLEMBOLA IS ALTERED BY RAINFOREST CONVERSION TO PLANTATION SYSTEMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript written by Susanti et al. studied trophic niche differentiation of Collembola in different forest types in Indonesia. The manuscript is well-written and can contribute to the trophic ecology of soil organisms.
Citing papers are enough, but need to be confirmed. E.g. Potanov et al. 2019 was cited in the text, but not found in references. Instead, Potanov et al. 2018, 2018a, and 2018b were found in references, which need to be identified. There are some other mistakes in forms, like (Korotkevich, Potapov, Tiunov, & Kuznetsova, 2018), (Josef Rusek, 1998) etc.
There are some small corrections needed as isotopic point of view (see General comments).
Experimental design: The manuscript used the study system that has already been used in several papers. The results are nearly the same as the previous papers, and the results seem to be rigorous.
Cite references to show the validity of using glycerol:water solution and 70% ethanol for the preparation of isotope measurement.
Validity of the findings: The authors studied Collembola species/genera among the vegetations. The isotope values are generally understandable among the vegetations. Results provide interesting information on tropical ecology.
Additional comments: There are some minor comments to the manuscript.
Line 170 The factor 1000 is an extraneous numerical factor and should be deleted (Coplen, TB (2011) Rapid Commun. Mass Spectrom. 25: 2538–2560).
Line 172 Vienna PD Belemnite is a strange expression. I suggest either Vienna Pee Dee Belemnite or VPDB.
Line 173 Multiple working standards should be used to calibrate isotope values (Skrzypek (2013) Anal Bioanal Chem 405: 2815-2823).
Lines 216, 314 and several places. The authors should not use “enriched” in “δ13C” of “δ15N”. Instead, I suggest either “All Collembola species were enriched in 13C relative to leaf litter…” or “All Collembola species had higherδ13C values relative to leaf litter…  (The authors used correctly in line 276 and several places.)


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: TROPHIC NICHE DIFFERENTIATION AND UTILISATION OF FOOD RESOURCES IN COLLEMBOLA IS ALTERED BY RAINFOREST CONVERSION TO PLANTATION SYSTEMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript was well structured in clear English. Introduction may need to be revised. I checked the raw data was supplied. Figures seemed sufficient.

I agree with the authors’ claim that there are few studies investigating Collembola food attributes in tropical ecosystems. However, I could not understand  what is the difference of soil ecosystem between Tropical and Temperate forests. Is there some difference of the soil systems for Collembola between tropical rain-forests and temperate forests? Please explain it.

Are there some problems of plantations in tropical forests, in ecological or economical sense?
Experimental design: Do you have species abundance data of the community?
Did the measured species number in Figure2-3 reflect to real species richness or community compositions?
Stable C-N isotope mapping at community level should have been biased in species selection processes, although the authors explained that species which were measured isotopes were covered 70% individuals (L159). If authors have the abundance data in each land-use, you can avoid the bias because of the direct evidence. Table of each plot only with dominant species abundance and total number may be sufficient for this purpose.

L130: Did herbicides directly decline some soil Collembola?

I think that pesticide, herbicides or fertilizer in management forests should disturb the C-N isotope ratios. Please explain it. (see also L323)

L183-207: Authors used the statistical analyses in three LMM models for land-use, species, taxonomy and lifefrom. Which was the most important factor for stable C-N isotope ratios?
I think that species identity should not be used as random effect in the first analysis, because species effect on stable isotope ratio should be important for the community isotope ratios.

L192: The range of delta C and N should be quantitatively evaluated using some criteria, such as the difference between minimum-maximum, functional diversity index (such as Rao’s quadratic entropy) or coefficient of variance (C.V.).
Validity of the findings: Interpretation of the results were described in intrinsic reasons, such as taxonomic difference and life-form difference in Collembola to land-use. However, as Figure 4 & 5, lifeform mapping and taxonomic mapping were not consistent with each other land-use type, and seemed not so robust the discussion about life-form and taxonomy in line of Potapov et al. (2016 Soil Biol Biochem).
I think that the changes by land-use in organic matter composition as habitat structure for Collembola, such as L, F, H compositions, may be important for Collembola community.  The variations of isotopes in taxonomy and life-form (Potapov et al. 2016 Soil Biol Biochem) may be context dependent. Please explain how the isotope variations changed by land-use in regards to life-form and taxonomy.
Collembola food items, represented to C-N isotopes, partly reflect to the humification gradients of the habitat substrate (Hishi et al., 2007 Soil Biol. Biochem), suggesting organic matter accumulation may relate with the ranges of the Collembola feeding items. Therefore, the ranges of C-N isotopes might be due to the range of humus gradient. If authors have the data, try to explain the variations of isotopes from accumulation of forest floor of each land-use.

Overall, data of this study of tropical Collembola was well structured, and useful for broad readers.
Additional comments: I agree with the authors’ claim that there are few studies investigating Collembola food attributes in tropical ecosystems. However, I could not understand  what is the difference of soil ecosystem between Tropical and Temperate forests. Is there some difference in soil system for Collembola in tropical rain-forest?


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: TROPHIC NICHE DIFFERENTIATION AND UTILISATION OF FOOD RESOURCES IN COLLEMBOLA IS ALTERED BY RAINFOREST CONVERSION TO PLANTATION SYSTEMS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall the manuscript is well written, in clear and professional English throughout - for minor corrections see general comments below. The article is well structured, figure 1 took me a while to understand the interpretation, but that is because it is a violin plot rather than a normal box plot - I do not think the authors should change it, but it might be worthwhile to include a more in-depth description of how the figure is to be interpreted, so greater understanding can be drawn without resorting to google! Figures 2 and 3 are poor picture quality and a little blurry - this is likely due to a low resolution inclusion when uploading, but again should be checked in proofs. Also the order of the individual species should be the same position across the landuses - Pseudosinella for example should be at the end for all, as in all habitats.
Experimental design: Experimental design is rigorous and suitable for publication. More emphasis should be included on the number of treatments (land-uses) within the abstract, as well as replication within the results/figures.

However, I am unsure why only "dominant" species were chosen for presentation of isotopic analysis - is this due to biomass of samples? Looking at the raw data, it looks like all Collembola had their isotopic composition assessed?

I also think the results would be strengthened if abundance of Collembola was included - the numbers are there in the raw data. What is the difference between the different landuses - there looks like there is a similar abundance, this is interesting and could be discussed. Also looking at the individual species may provide interesting discussion point - there are quite large differences in abundance between the different landuses for Pseudosinella for example.

Did species found in different habitat (e.g. Pseudosinella) have different isotopic results between habitats - any significant differences? I didn't see this stated in results.
Validity of the findings: This is an interesting and novel study that looks at tropical locations for isotopic analysis of Collembola species, with comparison to temperate systems, and other soil fauna. Although I am surprised they did not consider the work of Illig that focused on tropical montane rainforests and isotopic composition of the soil fauna (or the tropical papers that have cited this)?

The differences in Collembola life forms was a novel way of analyzing the data and provided a greater understanding of habitat differences. It was well described within the text.
Additional comments: A useful study that provides additional information regarding the trophic structure of the soil food web, focusing on Collembola within different habitats in the tropics.

Minor comments are as follows:
Abstract - it is not clear how long after conversion the oil and rubber plantations have been growing. Also it is unclear that there are four landuses being investigated.

line 58 - life not live
line 81-82 Further, food resources and trophic levels also vary among taxa, for example, Poduromorpha...
line 85 - sparse not virtually lacking
line 97 - add monocultures of after and and before rubber and oil palm


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: TROPHIC NICHE DIFFERENTIATION AND UTILISATION OF FOOD RESOURCES IN COLLEMBOLA IS ALTERED BY RAINFOREST CONVERSION TO PLANTATION SYSTEMS
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I have checked that authors properly responded to my comments. I have agreed with their explanations.


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: TROPHIC NICHE DIFFERENTIATION AND UTILISATION OF FOOD RESOURCES IN COLLEMBOLA IS ALTERED BY RAINFOREST CONVERSION TO PLANTATION SYSTEMS
Review round: 2
Reviewer: 2

Output:
Basic reporting: In response to the authors statements regarding earlier reviews comments:
They have completed the majority of changes suggested by reviewers. However I am unsure why they did not reorder the species of Collembola in figures 2 and 3 - these figures are supposed to indicate the variation in isotopes of the different species of springtail across the different habitats. The order they have the species in "based on mean values across land-use systems" highlights that all land-use systems have springtails with a similar range of isotopic compositions. Whereas keeping each species in the same position on the graph for each of the land-uses would indicate the difference in isotopic composition for that species.
Experimental design: In the rebuttal letter the explanation for use of "dominant species" is clear, understandable and a valid reason, however this is not made clear within the manuscript.
Validity of the findings: Surprised they have separated abundance and isotopic composition into two papers, rather than one strong one. The authors state they have included abundance data in the supplementary data, this doesn't really add to the paper if it's not going to be discussed within the paper alongside the isotopic results.
Additional comments: Minor comments are as follows:

line 122 first mention of Latin names of rubber / oil plantations, should be included in abstract and earlier in MS
line 139 use the chemical name for the product rather than the commercial
line 151-154 are acknowledgements not methods and should be deleted
line 372 typo


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: FORMALIN-INDUCED PAIN PROLONGS SUB- TO SUPRA-SECOND TIME ESTIMATION IN RATS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The journal requires to submit ARRIVE 2.0 checklist as a supplemental file.
https://peerj.com/about/policies-and-procedures/#animal-research
Please check and submit it.

Follows are point to point comments.

L(ine). 72
The study by Meck (1983) should be deleted here, because the study was for rats.

L. 73
"Involvement of different mechanisms in time perception" might be "Involvement of different mechanisms *in the modulation of* time perception". The assumed mechanism underlying the time perception is SET (scalar expectancy theory) or information processing theory by Gibbon (1984) in both short and long duration, isn't it? So, the "mechanism itself" is same in both durations. The difference is "in short duration, but not in long duration, the arousal effects on internal clock". Right?
Similarly, in line 78, the mean of "from" different aspects is unclear. What author's want to say is, as above, in short duration, but not in long duration, the arousal effects on internal clock"?
Taken together, the topic (first) sentence of the second paragraph might be such as "Considerable number of evidences suggest that the perception of short but not long duration is modulated by negative emotional stimulus" This is a phenomenon. After that, the description moves onto the mechanism, namely, the arousal modulates clock only in long duration.

L. 79
"attention and arousal are involved" is seems to be false. The Gibbon's model did not include these components and not refer to the effect of arousal or attention. Alternatively, Treisman (1963) --- Psychological Monographs: Gneral and Applied, 77, 1-31. --- refers to it. Treisman is more suitable here because the model by Gibbon is basically based on animal studies, whereas the Treisman's one is a traditional model for human timing and refers to the effect of arousal on pacemaker. So, bothe of Treisman and Gibbon should be cited for stating that the accumulator-pacemaker model, and Treisman and Noulhiane et al. are cited for stating the effect of arousal.

L. 86
One piece of logic seems to be skipped between the sentence "As a complex ... Freitas et al 2015)" and "A notable ...". The former paragraph said that arousal modulates clock only in short duration. The first sentence of this paragraph said that pain increases arousal. So the missing piece seems to be "Therefore, pain might modulate time perception (or clock) only in short duration." After that, "However, a notable problem is ...". I believe this revision will fulfill the chain of logic.

L. 89
If the paragraph is finished such as "Therefore, it is important to clarify the effect of pain on long as well as short temporal range", the connection to the next paragraph seems to be smooth.

L. 90
Again, the chain of logic between the first and second sentence is unclear. The first sentence means "human use counting in long temporal range". In natural way, "by contrast" to this, "the animal did not use counting". Nevertheless, the manuscript said "animals are often trained to perform the time perception task in a long temporal range". The sentence such as "In order to know the effects of arousal on longer temporal range, using animal is helpful because animal does not use such strategy" is needed.

L. 108
Please clarify the procedure of the handling (e.g., duration per day, number of times).

L. 110
Please clarify the extent of water deprivation. That is important information for animal welfare.

L. 115
You have declared the approved No. is H16036 on the on-line check at the submission of the manuscript and confirmed it. You uploaded the approval document as a supplemental file, but I could not find the number on it. Could you evidence the approved No. of this experiment is actually H16036? If you can evidence it, please show the No. in Method (The guidance require it, if the No. is available).

L. 117-123
The section of "Temporal bisection task" consists of only two sentences and contains not so much information.
I recommend delete this section and that the first sentence is moved onto the introduction (around L. 96?) and the second one to the procedure.

L. 119
I recommend to cite the first study using temporal bisection task too (Church & Deluty, 1977, J Exp Psychol Anim Behav Process, 3, 216-28.  doi: 10.1037//0097-7403.3.3.216.)

L. 124
Please enlarge the font size of the heading "Experimental apparatus" as the other heading such as "Animals".

L. 139
Although the term "training" is distinguished from "testing" in line 125, "test" is used to mean "session" in line 139. It is confusing. Please replace "test" in line 139 with "session".

L.168
The heading of "Experimental procedure" should be moved after the section of "Experimental apparatus". After that, the Headings of "Temporal discrimination training" and "Temporal bisection testing" should be put on the layer under the heading of "Experimental procedure" (namely, without new line), in order to clarify the flow of the experiment.

L. 144
Please explain the "anchor-duration".

L. around 144
Please clarify the ratio of short:long trials. Maybe 1:1?

L. 160
Please clarify not only the average but also the range of ITI.

L. 179
Is the number of paw licking and paw lifting counted within the drug-injected bisection task? With video-recording or real-time counting? The count was done with blinded? Please clarify.

L. 184
Please clarify the mean of "long response (PL)".

L. 185
Temporal bisection "task" is temporal bisection "session"? Please clarify that the curves were fitted to both individual rats (for analysis of PSE and WF, Fig1 F, G etc) and averaged psychophysical function (Fig.1D, E, etc).

L. 201
Here, change in the PSE and Weber fraction (WF) only were referred. Not only them, mean PSE and WF were calculated. Please describe them too.

L. 208
Many studies claim that Duncan's method is not valid as a multiple comparison (e.g., Hsu, 1996 attached file, etc.). Moreover, as far as I know, the standard way as a post-hoc analysis after the interaction is significant in ANOVA is test for simple main effect, lower parts of ANOVA (e.g., attached file). Please consider to use the simple main effect as a post-hoc analysis.

L. 209
0.05 (5%) is a criteria for significance. P is the calculated value from data. So α=.05, not p=.05.

L. 209
"Temporal bisection task trials with ..." is "The data from trials with..."?

L. 220
Please check the data of Fig. 1C. I calculate them and got 123.00 and 13.63 for formalin and saline, respectively. The graph looks unmatched to the values. Also, please check t value. I got 4.433.

L. 222
"goodness of fit" is more generalized term, so please use "R^2" or "coefficient of determination" here.

L. 222
0.94 is group mean? or minimum value? Please clarify.

L. 223
When is the "two days"? Baseline and drug challenge days? Pleases clarify.

L. 228
"Accuracy" has a specific mean in a context of timing study (that is, the discrepancy between the duration of to-be-timed stimulus and estimated "subjective" time or the length of the subjective time itself). Please replace it with "correct response rate".

L. 232-242
Please rewrite these parts. The Fig. 1D, E is separated by days (baseline and injection) and compare saline vs. formalin in each day. However, the text compares between days in each group. The reason why the control (saline) group is prepared is to compare it with experimental group. So, the important comparison is between groups. Therefor, two-way anova should be done with group by duration in each day. I think three-way anova is not so important here (in line 223). Do you delete it? For PSE and WF, post-hoc analysis should be done with simple main effects, if the interaction of two-way ANOVA (day vs. group) was significant, as mentioned above.

L. 315
Please write the values of statistical tests also for no significant difference in three days.

L. xx
In discussion, please refer to the persistence of pain throughout the session using Fig. 1A-C and Fig. 2A-C.

L. 351
The authors say that "believed to be ... motivation, attention, and decision-making" are influenced by pain, but insist that in this experiment, "motivation, the ability to discriminate these anchor duration and physical activity or attention were still normal" ... "we ruled out these factors". As the authors say, motivation and attention might be ruled out. However, how about "decision-making"? In the latter sentences, the term "decision-making" disappeared. Instead, "ability to discriminate these anchor duration", "physical activity" are introduced. Please clarify the relationship between "decision-making" and "ability to discriminate these anchor duration", "physical activity".

L. 384
What is the "distinct physiological characteristics"? In the manuscript, have the distinct physiological characteristics between short and long duration been referred until  here?. Also after this sentence, cognitive but not physiological explanation starts. Could you delete this sentence (, which may reflect ... stimuli.)?

L. 389
Should the animal studies addressing that attention modulates time perception  be cited? For example, Buhusi & Meck, 2006, J.Exp. Psy. Animal. Behav. Proc. 32, 329-338.; Buhisi & Meck, (2009) Phl. Trans. Royal. Soc. B,364, 1875-1885.) etc.

L. 389-391
What for is the evidence of the increase in latency here? Please clarify.

L. 396
Please give more detailed explanation for the term "pure".

L. 409
Kamada & Hata (2018; 2019) repeatedly reported that conditioned fear induced underestimation of duration. If you have some ideas to bridge this discrepancy, please mention it. If no idea, at least, please cite them like " ... , but see Kamada & Hata, 2018; 2019."

L. 424
What will occur if you continue the after-injection sessions? You implicates that the effect is conditioned response (CR) around line 407-409. If so, it is expected that prolonged after-injection training extinguish the CR. The last part of this paragraph seems to finish suddenly. I believe that the paragraph can be finished neatly If you add what will occur when the prolonged extinction sessions are given.

Caption of Figure 1
"Fitting curves for the proportions of ..." is "mean and their fitting curves"? (also the caption for Fig. 2).

That's all.


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: FORMALIN-INDUCED PAIN PROLONGS SUB- TO SUPRA-SECOND TIME ESTIMATION IN RATS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall, the paper is clear and unambiguous. They have sufficient references and background context. The article structure is professional, and self-contained.

There are a few small exceptions, most notably the Results section is difficult to follow, and the introduction/discussion could use a more careful description of the theory behind the paper, and the implications the findings have for the theory.
Experimental design: The experimental design fits within the aims and scope of the journal. The research question is (mostly) well defined, relevant, and meaningful. The work was does with high technical standard, and the methods were (mostly) described with sufficient detail.

There were a few small questions remaining about the methods section, noted in the review.
Validity of the findings: The conclusions are well stated, and the authors shy away from too much speculation. I did not see access to the data, but I may have simply missed it.
Additional comments: Overall, it's a fine paper that should be published in PeerJ in some form. I have a handful of comments for the authors in my longer review, attached 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: FORMALIN-INDUCED PAIN PROLONGS SUB- TO SUPRA-SECOND TIME ESTIMATION IN RATS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comments.
Experimental design: No comments.
Validity of the findings: No comments.
Additional comments: I highly appreciate for the authors exhaustive and reasonable reply.  Most of them convince me except for below minor three points.

1) Please forgive me for my poor explanations.

My original comment:
L. 201
Here, change in the PSE and Weber fraction (WF) only were referred. Not only them, mean PSE and WF were calculated. Please describe them too.

Reply: Accordingly, we described the mean PSE and WF in the Results section:

My reply to the authors reply:
My intension was as follows: Around L 197 of the revised version, the authors wrote only "we calculated the change in the PSE" but not about the mean PSE (and now I recognized also about Weber fraction). But I think the mean PSE is more basic index than change in PSE. In fact, the mean PSE was shown in Fig. 1 & 2. When I see the Fig.1 at first, I was wondering why change in PSE was not shown here even though the author wrote they calculate change in PSE. So, my requirement was "please write that the authors calculated the mean PSE (and also Weber fraction) as well as change in PSE here, around L197". On the other hand, I agree with the addition of the mean values of them for the Fig.4 in the RESULTS section.

2) About the ANOVA of (treatment) x (time)
I agree with the addition of the ANOVA that is correspond to figures. However, in L236, "(Fig. 1D)" and "(Fig. 1E)" is remained. These figures do not show " For each group, the differences of PLs between baseline day and injection day (L235-236)". Please delete the term "(Fig. 1D)" and "(Fig. 1E)" here (not the Figures themselves!)

3) L 280
When an interaction is not significant, we cannot move onto the test of simple main effect. Here the interaction was not significant so that the description in the text and asterisks at 5.04-s in Fig2E should be delete. Alternately, the main effect of treatment was significant on the injection day. Therefor, an asterisk may be added between the legend of the symbols (between " --o-- Saline" and "--o—Formalin").

That's all


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: PREDICTORS OF TREATMENT FAILURE DURING THE FIRST YEAR IN NEWLY DIAGNOSED TYPE 2 DIABETES PATIENTS: A RETROSPECTIVE, OBSERVATIONAL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this well-written, retrospective cohort study, Sia and colleagues investigated the clinical predictors of first-year treatment failure in 5579 Asian patients newly diagnosed with type 2 diabetes mellitus (T2DM). As demonstrated in the UKPDS trial, achieving early glycemic control can significantly reduce the long-term risk of microvascular and macrovascular complications in newly diagnosed T2DM patients, however, many uncertainties and controversies surround the possible predictors of poor glycemic responses in real life scenarios and further evidences are required in this field. Despite its interesting subject matter, unfortunately, the present investigation has major methodological flaws that would inflate the validity of the findings and predictive modeling.
Experimental design: 1. The study participants were enrolled across a very long-time window (Jan 2002-Dec 2017, 16 years), thus implying that available first-line pharmacological treatment options (e.g. metformin, insulin secretagogues, DPP4 inhibitors, SGLT2 inhibitors, acarbose, TZDs, GLP1RAs, insulin) varied considerably, depending on whether these newly diagnosed T2DM patients initiated treatment earlier or later in the study window. Other than a different glycemic efficacy, the occurrence of first-year treatment failure may reflect either a specific medication intolerance, that often prevents patients maintaining their therapy, or therapeutic inertia, which is a non-negligible problem among diabetes care providers. Failing to control for these confounding variables, and their combined effects, may have distorted the associations, and should be regarded as an important study limitation.

2. Beside pharmacological treatment options, also glycemic targets have varied considerably during this wide study window, so that the definition of treatment failure used (“never achieving post-treatment HbA1c <8% at 3, 6, 9, or 12 months after initiating treatment during the first year”, lines 94-95) appears misleading. It is true that before 2004, the American Diabetes Association (ADA) recommended that HbA1c not be allowed to exceed 8.0%. However, since 2004, the ADA dropped the traditional 8.0% “action threshold” in favor of a general recommendation to treat most patients to <7.0%, especially those newly diagnosed with T2DM and expected to get clinical advantage of metabolic legacy. Few exceptions to this general recommendation are represented by frail, older adults (> 65 years of age), in which less stringent HbA1c targets are recommended (8-8.5%) since late 2000s. I would suggest narrowing the study window to ensure more homogeneous criteria for treatment and outcomes of the study participants, redefining treatment failure, and excluding elderly patients with high Charlson comorbidity index scores from the primary analysis.

3. Although legitimate, backward stepwise regression modeling is particularly subjected to multicollinearity problems when the predictive variables are intercorrelated. The methods section should explain how multicollinearity was handled.
Validity of the findings: See the points above.
Additional comments: No additional comments


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

Input:
Title: PREDICTORS OF TREATMENT FAILURE DURING THE FIRST YEAR IN NEWLY DIAGNOSED TYPE 2 DIABETES PATIENTS: A RETROSPECTIVE, OBSERVATIONAL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: This paper focuses on an interesting topic. However, there is a major concern that must be addressed.
The authors stated that “Participants with at least one of the four post-treatment HbA1c levels <8% were categorized as non-TF (reference group)”. This critical choice must be strongly supported, otherwise it appears arbitrary and invalidates the study conclusions. Why didn’t they choose only the HbA1c value at the end of the observation? In my opinion this value is more appropriate as cut-off between TF and non-TF. In any case, please, show in table the HbA1c values at the four assessments.


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: PREDICTORS OF TREATMENT FAILURE DURING THE FIRST YEAR IN NEWLY DIAGNOSED TYPE 2 DIABETES PATIENTS: A RETROSPECTIVE, OBSERVATIONAL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: In their replies, the authors have satisfactorily addressed the major issues raised by this reviewer. The manuscript has been significantly improved and is now worth of publication. I do not have any further comments.
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: PREDICTORS OF TREATMENT FAILURE DURING THE FIRST YEAR IN NEWLY DIAGNOSED TYPE 2 DIABETES PATIENTS: A RETROSPECTIVE, OBSERVATIONAL STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: My concerns have been addressed, thus the paper is now susceptible for publication.