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: HIGH VARIABILITY OF DUNG BEETLE DIVERSITY PATTERNS AT FOUR MOUNTAINS OF THE TRANS-MEXICAN VOLCANIC BELT
Review round: 1
Reviewer: 3

Output:
Basic reporting: Improving the English used throughout the paper would greatly improve clarity. In particular the syntax of certain sentences make comprehension difficult:
•	Lines 24-26: Reword
•	Lines 31-32: Reword
•	Line 54: “refugees” should be “refuges”
•	Line 71: Reword this sentence
•	Lines 74-77: Reword this sentence to be more concise and make the overall point clearer
•	Line 82: missing degree symbol from “19”
•	Line 83-84: I’m unclear on the meaning of this sentence. Does “peak elevation” refer to the height of the eight highest volcanoes, the height of all mountains in the region, or the highest point of the highest point of the high plateau? To me “peak elevation” suggests the highest point of the plateau, but the fact that a range is given indicates on of the other two options.
•	Lines 94-96: Reword
•	Line 100: should be “Temperate”
•	Lines 100-115: This whole section could be reworded to make the overall point clearer
•	Lines 116-120: Reword this as its unclear just what makes this study unique. Is it the first study to look at dung beetle diversity solely in the TMVB? Is it the first to look at diversity between 2700 and 3400 MASL? Is it the first to look at diversity at these four mountains? Or is it some combination of these?
•	Line 135: should these elevations be 2700 and 3400 MASL?
•	Line 141: “…patches where trees are at an early….”
•	Lines 142-148: Does this paragraph refer to the sites in the TMVB?
•	Line 204: “…nearly the same across altitudes…”
•	Line 209: “…mountains…”
•	Lines 212: This sentence can be removed as the content is covered in the subsequent paragraphs.
•	Line 242: “…O. ch. Chevrolati…”
•	Line 264: “…These mountain communities…”
•	Line 294: “…O. ch. Chevrolati…”
•	Line 296: “…O. ch. Chevrolati…”
•	Line 201: “…O. ch. Chevrolati…”
•	Line 316-317: Reword this sentence

Minor notes
•	Lines 54-57: Provide references for this statement
•	Lines 100-102: move references to the end of the sentence
•	Lines 112-115: Provide references
•	Line 129-134: Including the elevations of all of the mountains would be good
•	Lines 142-148: Provide references
•	Line 165: reference Larsen & Forsythe, 2005 for trap spacing?
Experimental design: The methodology of the study is generally well described however the addition of some minor details would be appreciated.
•	Line 164: What months of the year does the rainy season occur?
•	Line 165: What type of dung was used?
•	Line 165: I’m a little unclear on how the trapping was repeated. Was trapping undertaken twice at each site, per year, e.g. in June 2001 traps at each of the four sites on MA were baited and left for 48 hours twice?
o	How far apart were these two sampling dates?
Validity of the findings: The content of the discussion is predominately speculation. While this speculation is relevant to the questions asked and well referenced there should be greater emphasis on addressing the stated aims of the study.
•	Lines 276-277: While this statement may be true without some more context regarding the number of pholephile species found outside of mountain areas it doesn’t seem to be borne directly from the results.
•	More discussion concerning the effect, or lack-thereof, of the ecological variables
•	More discussion on the biogeographic affiliation of the species and patterns of dispersion, given that these are emphasized throughout the paper
•	Some discussion on the temporal variation found would be welcome especially considering the large differences in the abundances of some species in 2011 compared to subsequent years seen in table S1

Minor notes
•	Lines 203-204: these percentages add up to 101%
•	Lines 206-207: can the specific volcanoes be given
•	Line 207: the fact that 13 species were found at only one of the four volcanoes is repeated later and can be removed.
•	Line 237: The abstract states that once O. ch. Chevrolati was removed from the analysis there was a positive relationship between distance and CS however in the results it states that CS decreased with distance.
Additional comments: The paper presents a number of new and interesting findings, is well referenced and the figures are clear and informative, however it suffers due to poor clarity and a lack of focus. In particular greater emphasis on describing and discussing the aims as stated in the paper would be good.


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: HIGH VARIABILITY OF DUNG BEETLE DIVERSITY PATTERNS AT FOUR MOUNTAINS OF THE TRANS-MEXICAN VOLCANIC BELT
Review round: 2
Reviewer: 1

Output:
Basic reporting: The clarity of the text has greatly improved from he previous version. I congratulate the authors for the effort.
Several references have been added to support some statements and the overall importance of the study as filling a knowledge gap.
Figures, tables and raw data comply with the journal standards.
Results are relevant for the aims and predictions.
Experimental design: Aims of the study meet the scope of the Journal.
This second version defines the aims of the study much better than the first version, fulfilling one of the main weaknesses pointed out by the three reviewers.
Methods are explained with enough detail for the study to be reproducible.
Validity of the findings: Data is statistically sound and perfectly controlled with several tools, perhaps even to much after reviewers demand.

Speculation in the discussion (a point in which the three reviewers commented) is much better supported with literature and results in the study.
Additional comments: NOT to be taken as a fault in this study but as a scientific thought:
I continue to wonder if other sampling sites could have been chosen (or could be studied in the future if more funds are available). This idea comes to my mind specially thanks to the greatly improved figure/map including elevation of the whole Volcanic Belt. The map clearly shows that there are at least 7 isolated mountain ranges in the Volcanic Belt, of which only 2 are sampled in this study. Moreover, these are the most eastern ones, leaving 5 mountain ranges not sampled.
I wonder if the patterns found in this study can be generalized to the whole Volcanic Belt or even if a more homogeneous pattern would be found (dropping the idea of the "high variability" mentioned throughout the manuscript and even the title).
I reinforce the idea that my suggestion does not lessen the value of this study, it just expresses my eagerness to see this research continue with further sampling in the other mountain ranges in Mexico (and elsewhere).


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: HIGH VARIABILITY OF DUNG BEETLE DIVERSITY PATTERNS AT FOUR MOUNTAINS OF THE TRANS-MEXICAN VOLCANIC BELT
Review round: 2
Reviewer: 2

Output:
Basic reporting: I acknowledge the efforts of the authors to include in the new version most part of my former suggestions and criticisms. In the current version I only have some minor proposals now included as notes in the PDF version of the manuscript.  If the authors agree to them, I suggest the acceptance of the manuscript without the need for a subsequent revision.
Experimental design: I now only suggest alternative definitions of vertical and horizontal colonizations
Validity of the findings: OK
Additional comments: I acknowledge the efforts of the authors to include in the new version most part of my former suggestions and criticisms. In the current version I only have some minor proposals now included as notes in the PDF version of the manuscript.  If the authors agree to them, I suggest the acceptance of the manuscript without the need for a subsequent revision.


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: HIGH VARIABILITY OF DUNG BEETLE DIVERSITY PATTERNS AT FOUR MOUNTAINS OF THE TRANS-MEXICAN VOLCANIC BELT
Review round: 2
Reviewer: 3

Output:
Basic reporting: Some minor typos:
•	Line 88: reference
•	Line 277: missing “of”?
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The readability and flow of the paper is greatly improved. The introduction is well referenced and provides a good overview of the concepts underpinning the research as well as the important geographical context. Holes in the literature are identified and the importance of filling these holes well justified. The results of the study lend support to many of the hypotheses put forward and demonstrate the importance of sampling effort when surveying montane communities. The high degree of variability between communities combined with the dominance of O. ch. chevrolati was extremely interesting, particularly as it was found in both forests and pastures.


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: PSYCHROPHRYNELLA GLAUCA SP. N., A NEW SPECIES OF TERRESTRIAL-BREEDING FROGS (AMPHIBIA, ANURA, STRABOMANTIDAE) FROM THE MONTANE FORESTS OF THE AMAZONIAN ANDES OF PUNO, PERU
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript has good flow, professional English and is clear. But, the Figures (especially 1 and 3) must be improved, in Fig. 1 I suggest that the legend of species be included within of the Figure and reduce the size of the elevation legend. In Figure 3 is need improve the quality of photographs, the edge of fingers and toes are poor quality and between Toe II and Toe III is possible observe noise it looks like webbing.
Experimental design: As mentioned in the manuscript (attached) for me the major problem is the generic assignment, I suggest that the authors perform a phylogenetic analysis with the genetic data that they obtained and with those available in GenBank for Psychrophrynella and Microkayla, especially for demonstrate that the new species is not Microkayla
Validity of the findings: Linked to the generic assignment the finding of the authors is weakened due that is unclear if the new species is Psychrophrynella or Microkayla.
Additional comments: Is clear that the author have a new species in hands and that it need be described, but the manuscript should be improved in several aspects before of be published.


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

Input:
Title: PSYCHROPHRYNELLA GLAUCA SP. N., A NEW SPECIES OF TERRESTRIAL-BREEDING FROGS (AMPHIBIA, ANURA, STRABOMANTIDAE) FROM THE MONTANE FORESTS OF THE AMAZONIAN ANDES OF PUNO, PERU
Review round: 1
Reviewer: 2

Output:
Basic reporting: The English usage is fairly good.

The structure, literature cited, methods, results and discussion are all properly arranged, comprehensive, appropriate, and clear. Figures are relevant, necessary, and properly labelled.
Experimental design: Being a basically descriptive work, no experimental design was required. Methodology is appropriate and pertinent standards for its different sections (morphometry, genetics, bioacoustics) have been followed.
Validity of the findings: Besides the intrinsecal interest in the finding of a new species, the extension range of the genus Psychrophrynella to the Cordillera de Carabaya points to this region as a hotspot for Holoadeninae diversity and radiation, which represents one of the most interesting contributions of this paper.
Additional comments: The current manuscript presents an uncomplicated description of a new species of frog in the diverse  clade Holoadeninae, which is showing an astonishing increase in species descriptions in the last years. The English usage is fairly good, although I (a non native English speaker either) have made some corrections and suggestions in the MS itself, to improve it. This, together with some comments, I recommend to address in order to fix some unclear things and improve the MS's general comprehension.

Regarding the References section, I wonder whether citation of Pie et al. 2017 is accepted by PeerJ as a valid reference, considering that it is not a publication but a non peer-reviewed piece of text posted at a website.


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: PSYCHROPHRYNELLA GLAUCA SP. N., A NEW SPECIES OF TERRESTRIAL-BREEDING FROGS (AMPHIBIA, ANURA, STRABOMANTIDAE) FROM THE MONTANE FORESTS OF THE AMAZONIAN ANDES OF PUNO, PERU
Review round: 1
Reviewer: 3

Output:
Basic reporting: This is a very good species description. I have only found a few minor issues that are marked in detail in the attached pdf file.
Experimental design: This is not an experiment driven research, but a descriptive and comparative study. The research is rigorous, sound, and following required ethical and legal aspects.
Validity of the findings: The authors present good quality and unambiguous evidence that they have discovered a new species. Conclusions fit well with data and analyses. The only exception is the advertisement call. The authors state the recorded a single call from an unvouchered specimen but they fail to explain how they assigned the recorded sound to the species. Furthermore, at times, the authors talk as if they have recorded more than one call.
Additional comments: Congratulations. Very good manuscript. My comments are mainly cosmetic.


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: PSYCHROPHRYNELLA GLAUCA SP. N., A NEW SPECIES OF TERRESTRIAL-BREEDING FROGS (AMPHIBIA, ANURA, STRABOMANTIDAE) FROM THE MONTANE FORESTS OF THE AMAZONIAN ANDES OF PUNO, PERU
Review round: 2
Reviewer: 1

Output:
Basic reporting: As previously I noted the manuscript has good flow, professional English and is clear. Nevertheless, I suggested review the figures 1 and 3. The authors rejected review the figure 1 arguing that "the suggested improvements to represent personal preferences of the reviewer", maybe it is true however also indicates that the authors don`t like improve the manuscript and they prefer use a template as figure 1 that is identical to the used in Catenazzi et al. 2017. ZooKeys 685: 65-81 except in the symbols of the species. Also, the authors say "We have improved the quality of Figure 3" but only erased the noise between Toe II and Toe III, really they did not take new photos.
Experimental design: As mentioned previously for me the major problem is the generic assignment, I suggested that the authors performed a phylogenetic analysis with the genetic data that they obtained and with those available in GenBank for Psychrophrynella and Microkayla, especially for demonstrate that the new species is not Microkayla. I am not expert in these clades but consider that the phylogenetic tree improve the manuscript, I agree with the authors in that they have a new species of Psychrophrynella but is really important that they perform the phylogenetic analysis. If the authors reject perform the tree they should eliminate the genetic distances as generic assignment because the genetic distances are only adequate to match specimens to groups, not for species delimitation, and even less for trying to assign specimens to clades. Phylogeny and divergence is not the same. You can be very divergent from your sister group, and yet be the sister group, or can be very similar to a very distant relative.
Validity of the findings: Linked to the generic assignment the authors could improve the manuscript if perform a phylogenetic tree. Also, add a phylogenetic tree establish a precedent in the knowledge of these genera and maybe for the widespread of phylogenetic trees in future taxonomic works for these genera.
Additional comments: I consider that the author have a new species in hands but is important that the authors improve the manuscript, all the comments in review were for improve the manuscript and never negatively but the authors really not make a significant effort for add the comments and suggestions.


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: IMPACT OF PARENTS’ PHYSICAL ACTIVITY ON PRESCHOOL CHILDREN’S PHYSICAL ACTIVITY: A CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Using a regression analysis, the current study examined the associations of PA level (i.e., sedentary behavior, total physical activity, and moderate-to-vigorous physical activity) between parents and their pre-school children based on continuous time spanning five weekdays and two weekends, using an ActiGraph GT3X accelerometer. Results revealed a significant correlation between mothers’ and girls’ moderate-to-vigorous physical activity (MVPA) and total physical activity (TPA) on weekdays, while no such effect was observed between fathers’ and children’s TPA. In addition, Father’s MPVA was found to be correlated with those of boys’ and girls’, with paternal influence appearing to be stronger than maternal influence. The results indicate that parents’ PA may be associated with their preschool children’s physical activity level, with maternal influence stronger during the weekdays and paternal influence stronger on the weekends. Manuscript was written well with professional English throughout. Most of the literature references are provided sufficiently. However, the reviewer was expecting a more informative literature review on how the ActiGraph GT3X has been used and considered as a better tool compared to physical activity questionnaire and in-person observation. A paragraph to justify the measurement will be suggested. Specifically, after the brief description on the ActiGraph GT3X, on Line 93-94, the authors swiftly concluded their introduction and literature review without an adequate hypotheses.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The data collected from ActiGraph GT3X seems accurate and promising. However, the reviewer was curious if the working style from parents (i.e., sedentary versus non-sedentary working style) was entered as a controlled variable in the regression model. With the parents wearing the accelerator from 6 am to 11 pm every day, would the amount of physical activity measured simply reflect the nature of the parents' working style? Would a non-sedentary working style result in a more sedentary behavior after work for the parents?  In addition, recording data from 6 am to 11 pm seems insufficient to the reviewer as parents might have diffident work time, while a 9 am to 5 pm time period for data collection may provide a more standard time frame for measuring physical activity and total activity amount. The reviewer was hoping that the authors could provide an explanation on how the time frame was determined in the current study.


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

Input:
Title: IMPACT OF PARENTS’ PHYSICAL ACTIVITY ON PRESCHOOL CHILDREN’S PHYSICAL ACTIVITY: A CROSS-SECTIONAL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Generally, this is an interesting paper and the ideas flow logically. But the introduction needs more detail and I suggest that you improve the description at lines 73-80 to provide more justification of the background information (adding the literature references).
Experimental design: The topic is timely and will be of interest to the readers of the journal. But the study design was not new and the statement of how this research filled the knowledge gap was not made clearly enough. I suggest that you provide more justification for the study design.
Validity of the findings: The authors did not justify whether the assumptions for linear regression was met before applying the model and this may result in Type I or Type II errors. In addition, although some confounders were controlled in the model, other factors like the environment of the neighborhood were not on the list. This may affect the conclusion of this research. Besides, the effect size was not reported.
Additional comments: Overall, the topic is timely and will be of interest to the readers of the journal. But numerous statements are made that reflect the authors’ opinion but they are portrayed as fact with no references cited or not sufficient evidence provided. I suggest that the authors improve the strength of the statistical analysis before making the final conclusion.


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: IMPACT OF PARENTS’ PHYSICAL ACTIVITY ON PRESCHOOL CHILDREN’S PHYSICAL ACTIVITY: A CROSS-SECTIONAL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have successfully addressed the issued that the reviewer provided. The manuscript now is in a good shape and congratulate for the authors for the accomplishment!
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: REGENERATIVE AGRICULTURE: MERGING FARMING AND NATURAL RESOURCE CONSERVATION PROFITABLY
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript is clearly written with simple appropriate language. The references are adequate throughout  and the figures, raw data and structure are all reasonable.

I would not that in figure 2 there is no description for those data represented by the white bars
Experimental design: This research is field based and down from various sources on a farm level. Given the nature of the data used, there is an inherent limit to what how much actual experimental design and manipulation is possible. Within those confines, the authors did a commendable job of gathering their data. The methods for collecting insects etc. are quite sound and the ability to locate 20 farms and gain the relevant information from each makes for quite reasonable sample sizes.
Validity of the findings: The findings of this study are quite sound and reasonable given the data and analyses. The conclusions are sound and within the scope of those data collected. There is some reasonable speculation, and while this study is limited in its ability to provide mechanism, the speculation is aimed at these mechanisms which is appropriate.
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: REGENERATIVE AGRICULTURE: MERGING FARMING AND NATURAL RESOURCE CONSERVATION PROFITABLY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Acceptable
Experimental design: The paired design is implied in Table S1 but not clear in the Methods. Please clarify statistical design. See comments on the manuscript (attached).

L 74 - how were fields selected on each farm? Why were 2 omitted?
L 81 - how were the sites elected for yield determination?
L 83 - how was revenue for livestock grazing determined?
L 86 - Presumably numbers of indiv from all spp are mentioned. Clarify.
How many of each sp? Did 1 sp do more damage than another? Was crop damage due to pests assessed?
L 116-17 - how was the value of other products assessed?
L 118 - Extra cost only for seed or also for direct applications of insecticide?
L 123 - how many regen farmers received an organic premium? Show in T S1
Validity of the findings: T S2 - more explanation needed in order for table to stand alone. Stats related to T S2? Not referred to in text
Fig 2 - see comment bubbles on Fig 2. It's not clear to me what this shows. It appears that some costs are missing.
Additional comments: This type of systems analysis is required and appreciated.
Please address comments above in order to help readers understand how data were collected and analyzed.


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: MERGING CRANIAL HISTOLOGY AND 3D-COMPUTATIONAL BIOMECHANICS: A REVIEW OF THE FEEDING ECOLOGY OF A LATE TRIASSIC TEMNOSPONDYL AMPHIBIAN
Review round: 1
Reviewer: 1

Output:
Basic reporting: The submitted manuscript presents an interesting approach of combining histological studies with computational biomechanical analysis using FEA. I find this approach intriguing as it can help validate results and inferences obtained from FEA studies. However, the way this is presented I this manuscript has several issues:
The introduction is detailed, but on many occasions unnecessarily long. In particular the “TESTING FEEDING” section, which spans eight pages, reads more like a review of the specific topics and would be suitable for a PhD thesis. Given that methods, such as FEA are now applied routinely in palaeontological studies, this is unnecessary and just distracts from the novelty of the methodological approach. I would recommend condensing the whole section to no more than two pages, bringing out the advantages of the presented combined methods based on how these techniques have been applied separately so far.
My biggest concern with this manuscript is the lack of detail regarding the methodology. This is particularly evident in the description of the FEA on which eight lines were spent (in comparison to the introduction of over eight pages as mentioned above). I do understand, that the author refer to a previous publication, in which the generation of the FE meshes, loading scenarios and boundary conditions are laid out in more detail. However, this study should be self-contained and the reader should not be forced to hunt down another publication to understand what has been done. Furthermore, four different loading scenarios are discussed in the results and the discussion, but only three are figured. It is very unclear what the differences in model setup between the unilateral bite and the lateral bite are, for example. Similarly, details of the model construction are not given. This is particularly important to understand, if the patters observed in the FEA models in relation to the microstructure and suture morphology are realistic or if the model has been simplified to a where these similarities are purely coincidental. As the authors state several times in the manuscript, model simplification and geometry are important factors. I would therefore expect the methods section to be very detailed.
The presentation of the whole study is OK. I have made suggestions in the attached PDF regarding spelling and grammar, but would strongly recommend to have the manuscript proof-read by a versed English or native speaker. In the results section, different tenses are often mixed throughout. Be consistent here.
Presented figures are for most parts informative and I really like how the results are presented in figure 4. I would recommend combining figures 2 (at least C and D) in some form with figure 3 so that the position and orientation of the thin-section can be recognised without going back and forth between both figures.

Overall, I think this manuscript has lots of potential and could make and important contribution as it describes a new methodological approach for biomechanical studies. That's why it is even more important to provide all necessary details to understand and possibly replicate the study design. This is not yet the case, however, and I have recommended major revisions. Fixing the raised issues shouldn't be too difficult, though.
Experimental design: The general research question is well-defined and clear. It offers a new approach to supplement biomechanical studies and to infer palaeobiological properties. Knowing the work of some of the authors I assume the experimental setup to have been performed rigorously. However, this is not visible from the described methodology (see detailed comments above). A thorough explanation of the model generation and FEA setup is very much required.
Validity of the findings: I find that the findings are mostly supported by the results. However, it is difficult to ascertain this in some aspects, as the methodology is not described in sufficient detail. This particularly relates to the setup of (apparently) four different loading scenarios, while only three are discussed and figured. This seems to be a bit cherry-picking in only showing the results that match with the histological analysis. As criticised by the authors, only a small number of different scenarios is usually tested in FEA studies. Therefore, please, figure and discuss al scenarios here as well.
Additional comments: A few further minor comments in addition to those in the attached PDF:

•	Page 10, lines 102-104: I think this Is not true. A number of (biomechanical) studies have pooled various lines of information and evidence to complement functional analysis.
•	Page 11, line 131: “very promising attempts” – what makes those studies promising? Did those studies fully resolve the question about feeding mode and behaviour? I would consider rephrasing this to something along the lines of “…a number of studies have used computational biomechanical analyses to address the question of…”
•	Page 11, line 138: “correctness” is strong and in my opinion inappropriate word here. The correctness could only be tested if a living temnospondyl was biomechanically examined (and even then it might be difficult). Rather use “replicate the in-vivo behaviour” or similar.
•	Page 20, lines 324-325: Provide institutional abbreviations directly on first occurrence. It is mentioned later in the text, but the reader shouldn’t have to search for it.
•	Page 20, lines 324-329: Provide details on CT scan settings.


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: MERGING CRANIAL HISTOLOGY AND 3D-COMPUTATIONAL BIOMECHANICS: A REVIEW OF THE FEEDING ECOLOGY OF A LATE TRIASSIC TEMNOSPONDYL AMPHIBIAN
Review round: 1
Reviewer: 2

Output:
Basic reporting: The introductions section is very long (11 manuscript pages). I have made some suggestions of background information that could be cut/reduced, for example information not directly relevant to this study (why 12 citations on sauropods and marine reptiles demonstrating they have no extant analogues?) and other sections (introducing FEA and overview of general factors impacting bone strength) what can be shortened.

On the other hand, some very relevant findings from previous studies involving sensitivity analyses in FE studies of other fossil taxa and/or validation studies on FE models of crocodilian skulls (similar in both shape and ecological niche to temnospondyls) are not mentioned at all. Similarly, although correlating FE results with histology is novel, doing so with cranial sutures is not (Line 137) – see extensive work by Rayfield, Moazen Reed and others in fossil and living reptiles and fossil tetrapods.

It is unclear to me (see comments Line 304) why an aquatic predator cannot be an ambush predator using a lateral strike, as this occurs in some extant aquatic predators.

I have made suggestions (see “Validity”) on one figure that can be modified slightly and one figure that could be added that would clarify results.

The English language needs to be improved throughout the manuscript to ensure your international audience can understand your text. I suggest having an English speaking colleague review your manuscript. I have made some corrections and highlighted difficult text (any highlighted text without additional comments are areas where the language is unclear) throughout the annotated PDF.
Experimental design: One of the issues with this manuscript is a lack of clear questions that will be addressed or hypotheses that will be tested (stating how results from FEA and bone microstructure would support or reject those hypotheses). You must add such a section at the end of your introduction.

I realize that the detailed methods for the model have been published elsewhere but basic information needs to be repeated here – see annotated PDF line 371. This is especially true if – as you suggest – you have made changes to your model since its 2017 publication. Any such changes need to be detailed in full to allow the study to be replicated.

Your text (Lines 529 – 531) suggests the lateral strike loading regime is not the same as a unilateral bite. If not, please explain (early on when you discuss results from your 2017 paper) what a lateral strike load regime actually involves and how it differs from unilateral biting.
Validity of the findings: One thing that would help tremendously in understanding the Discussion section “Bone histology vs computational biomechanics – how does bone structure correlate with Finite Element Analysis?” would be if you could overlay bone boundaries onto the screenshots of FE models showing von Mises stress (Figure 8). Readers could compare these with the labelled maps of the skull shown in Figure 2 to understand within which bones different stresses are occurring under different loading conditions.

Also – why not have contour plots of compressive vs. tensile stress to see whether your predictions on stress type based on sutural morphology corresponds with FEA predictions? Von Mises stress plots do not add information beyond stress magnitudes, whereas suture shape should correlate with the type of stress (tension, compression) being experience.

Be careful with the language you use in the final section of the discussion (new interpretation of mode of life) – although you have evidence to support your suggestions, they are still speculation. Some of the wording you use makes these ideas sound conclusive.
Additional comments: The study being proposed here – in terms of using bone microstructure as a reality check on the predictions of finite element models – is genuinely novel, interesting and worthwhile, and I would like to see a revised version of this paper in print. There are some findings that I see as particularly interesting that should be highlighted more - particularly the impact of the deep and extensive lateral canals on skull mechanics in Metoposaurus and bone microstructure/FE results indicating that some bones are resistant to stresses incurred during head raising (rather than biting), which is not a commonly tested load regime.

However, there are numerous issues (see above) with the manuscript that need to be overcome prior to 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: MERGING CRANIAL HISTOLOGY AND 3D-COMPUTATIONAL BIOMECHANICS: A REVIEW OF THE FEEDING ECOLOGY OF A LATE TRIASSIC TEMNOSPONDYL AMPHIBIAN
Review round: 1
Reviewer: 3

Output:
Basic reporting: The paper contains some interesting data on bone histology and how histological features change across the skull of the Late Triassic temnospondyl Metoposaurus. However, I found the main aim of the paper was difficult to determine. What is the hypothesis/hypotheses being tested here? Is it that cranial histology can be used to predict skull stress? Is it that increased bone compactness and increased bone thickness are related to increased skull stress? Is it that cranial sutures can be used to predict cranial strain? Is it to test the null hypothesis that Metoposaurus was an active hunter, or passive bottom dweller? The hypothesis / hypotheses being testing need to be clear from the start of the paper.

The introduction is very long. It would benefit from sharpening and tightening the text. This will be made easier if the hypotheses are clearer. Also there are sections of text that are basic background information that don’t need to be included in a manuscript – a good example is lines 192 – 227 on bone structure. It would be appropriate to include such information in a PhD thesis for example, but it’s just not necessary to include such detail on basic bone biology.

Unfortunately, although the introduction is long, the authors fail to cite some key papers in the field. On line 136 it is stated that “Thus in the present study is for the first time correlation of histological and cranial suture morphology results with computational biomechanics (FEA) is done”. This is simply not true. There are lots of papers that compare histology and suture morphology to the results of FE-analysis – work by Moazen, Porro, Jasinoski, Rayfield on extinct and extant taxa – there’s a long history of papers in the this area but different research groups and they are not cited at all here. Some examples are: Rayfield 2004, 2005; Jasinoski et al. 2010a,b, Jasinoski & Reddy 2010; Moazen et al. 2009, 2013; Porro et al 2013, 2014; Curtis et al. 2013; Jones et al. 2011) . These papers need to be acknowledged and the statement rectified.

The paper would benefit from a check on the English language by a native speaker – see quoted text above as an example. There are many sentences that require rephrasing. This is concerning because I’m not aware if PeerJ editorial team will make such changes in house.
Experimental design: The authors outline why they are focusing on Metoposaurus, but if the aim is to demonstrate how cranial histology can be used to predict skull stress and strain, then why not complete the study on an extant animal where cranial strain can be measured experimentally ex vivo or in vivo, and then compare these measurements to bone histology? Whether the authors need to address the issue of why extant datasets were not used depends on what this paper is trying to achieve. If it is to document histological features across the skull and compare these to FEA results, then fine. But if it is to introduce bone compactness and thickness as a proxy for predicting skull stress and strain, then the authors need to address why they have not considered used extant animals as a case study.

Bone thickness and bone compactness are measured. However, on what basis do these two variables, either independently or combined, act as useful indicators of bone loading? Is this based on common sense logic – bones should be thicker and more compact where stresses are higher in magnitude? Or is this based on quantified data from living animals and associated published work? Please state why you used these metrics, and why they are appropriate stress indicators?

FE models. The FE models used in this paper have be published previously. However this manuscript needs to include much more information on model construction. What were the boundary conditions of the model – loadings and constraints? What material properties were used? How many elements etc. Yes, a reader can refer to the previous paper, but this current manuscript needs to contain sufficient information to be a standalone publication.
Validity of the findings: Sutures. If all the sutures in the skull are recognised as interdigitated, then why go to the effort here to talk about different types of sutures? And how does suture morphology link to skull strain? Granted, the von Mises plots will show the location of peak strain in this homogenous and linear elastic model, but von Mises doesn’t tell you whether the stress and strain is tensile or compressive. Do interdigitated sutures in the skull of Metoposaurus follow the predictions for the types of strain experienced across interdigitated sutures? IE are all the strains compressive across the interdigitate sutures? If not, then where does this leave the role of sutures as predictors of cranial strain? Do we believe the signal from the sutures or the signal from the models?

I felt that similar ambiguity was present when discussing the results of the comparison between bone compactness and thickness and skull stress. Figure 8 nicely illustrates the spatial relationship between FE-derived stress and microanatomy-derived stress, but I was looking for a conclusion to this work – which of these two metrics is the better proxy for biomechanical behaviour? But again, this relates to my comments at the start of the review – what is the manuscript trying to achieve? Is it trying to compare these two methods for determining skull stress? If so, how do you reconcile when the two types of data give conflicting results. Or is it attempting to work out the feeding behaviour of Metoposaurus? The discussion focuses more on the latter issue.
Additional comments: My overarching comment to the authors is that they need to specify what this paper is trying to achieve. Once this has been determined, everything else will flow much better – superfluous information can be cut from the introduction and the results and the discussion can be targeted better. And the authors must acknowledge the large body of work that has already attempted to link suture morphology to FE-derived stress and strain patterns.


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: MERGING CRANIAL HISTOLOGY AND 3D-COMPUTATIONAL BIOMECHANICS: A REVIEW OF THE FEEDING ECOLOGY OF A LATE TRIASSIC TEMNOSPONDYL AMPHIBIAN
Review round: 2
Reviewer: 1

Output:
Basic reporting: I find the revised version very much improved in comparison to the original submission. Suggestions and comments from myself and the other reviewers have been incorporated into the revised manuscript making the whole study more accessible to the reader.
Experimental design: See above
Validity of the findings: See above
Additional comments: I have a few minor comments and remarks listed below: In addition I have made some grammatical suggestions in the attached document in the form of track changes. The language of the manuscript has considerably been improved, but if possible another check by a native English speaker would be recommended.

Abstract

•	“To test this scenario, the early tetrapod group of Temnospondyli is an excellent case-study” – Some brief explanation is necessary here to support that statement.
•	“However, microscopic data suggest that…” – What is meant by ‘microscopic data’ here and on other occasions in the text?
•	Results in the abstract are very much focussed on the reconstruction of the palaeobiology of Metoposaurus, but there is barely any word about the methodology and how reliably histology can replicate the FEA results and vice versa. The latter is, in my opinion, the more interesting subject of the manuscript and should be mentioned in more detail in the abstract.
Introduction

•	“For fossils taxa without modern analogues these variables moreover have to be assume based on the taxa with a similar Bauplan, but sometimes far phylogenetically (Anderson et al., 2012) as it is i.e. for Temnospondyli where commonly the Crocodylus is used as a proxy (see Fortuny et al., 2016 for a discussion).” – It is not clear what is meant by ‘far phylogenetically’. Consider simplifying and rep[hrasing that sentence.
•	“It drive to make assumptions regarding bone properties and moreover calculations are simplified (see Bright, 2014; Fortuny et al., 2015 and references therein for discussion ).” -  Not clear what ‘it drive’ means. Consider rephrasing.
Discussion
•	“A slight increasing of stress level in the FEA model of skull raising (Fig. 7D reinforces the support to this hypothesis” – Rephrase sentence, meaning not clear.

Figures
•	Figure 2 caption is missing indices (A, B, C) for the different scenarios. In addition more detail is necessary explaining the different parts of the figure (e.g. load forces and scenarios)


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: MERGING CRANIAL HISTOLOGY AND 3D-COMPUTATIONAL BIOMECHANICS: A REVIEW OF THE FEEDING ECOLOGY OF A LATE TRIASSIC TEMNOSPONDYL AMPHIBIAN
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have made substantial improvements to the manuscript since the first version and they are to be particularly commended on greatly reducing the overall length of the Introduction (especially the FEA section) and improving it’s focus, adding clear study goals at the end of the introduction, the much more detailed methods section – particularly the careful description of the different loading cases and Figure 2, which is excellent and very clear, which muscles were incorporated into the model, and the nature of boundary conditions and material properties – as well as the excellent new Figure 7.

There are still minor revisions needed prior to publication.

The introduction could have a more orderly flow – at the moment, the authors 1) introduce FEA and simplifications that must be made in fossil taxa such as Temnospondyls, 2) introduce bone histology and how it may be related to mechanical loads, 3) introduce the most common bone histology found in temnospondyls, 4) formally introduce temnospondyls, 5) review previous FE studies on temnospondyls, 6) discuss more bone histology in temnospondyls, and finally 7) discuss study goals. It may be more straightforward to rearrange the sections, for example: 1) introduce temnospondyls and uncertainties of their feeding modes, 2) introduce FEA, review previous FE studies on temnospondyls, and discuss assumptions that must be made for fossil taxa, 3) introduce the idea that bone histology and feeding loads are related and discuss specific histology (and what this might mean for feeding loads) in temnspondyls, and 4) finally bring it all together in the study goals – they will compare data from FEA and bone histology to test the accuracy of predictions from FEA and try to uncover feeding mode.

The language in the introduction, results and discussion still needs improvement to ensure your international audience can understand your manuscript. I have made corrections until the end of the Introduction section of the text.
Experimental design: On several occasions (in the Abstract starting line 34, Introduction starting line 136) the authors refer to the taxon as an “excellent case study”. This is somewhat of an exaggeration – as pointed out by Reviewer 3 during the first round of reviews, it would be ideal to test out this idea on extant animals first. Metoposaurus is interesting because it is very common (and thus there are specimens to be sectioned) and its feeding mode is debated – I think the language here just needs to be toned down a bit.

The FE methods sections is greatly improved. One important detail that is still missing are the magnitudes of the applied muscle forces. These could be stated in line with the text, for example at Line 225 before the end of the sentence you could add: “…this resulted in applied muscle forces of XX Newtons for the AMI, XX Newtons for the AME and XX Newtons for the AMP.” This is important data in order to replicate the study. Also, what were these magnitudes based on – reconstructed cross-sectional area?

One last question – wouldn’t the metoposaur need to also bite down (unilaterally) on its prey during a lateral strike, in addition to pulling the prey sideways? Maybe add a line to justify modelling a lateral pull without the animal unilaterally biting down.
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: CRYSTAL STRUCTURE CORRELATIONS WITH THE INTRINSIC THERMODYNAMICS OF HUMAN CARBONIC ANHYDRASE INHIBITOR BINDING
Review round: 1
Reviewer: 1

Output:
Basic reporting: The present manuscript analyzes the correlations between thermodynamic signature of CA inhibitor binding and the corresponding 3D-structures of the resulting CA inhibitor complexes. Pairs of protein-ligand complexes were analyzed and divided into two groups of similar and dissimilar binding poses. From this similar pairs were found to be primarily dependent on entropy changes as a consequence of desolvation thermodynamics. In contrast, in dissimilar pairs, the compounds bearing large substituents bound more enthalpy driven, which was explained by the formation of hydrogen bonds to the hydrophilic part of the active site. It was also hypthesized that additional hydrophobic groups could complicate and potentially slow down the dissociation process. The study includes 7 new crystal structures of CA-inhibitor complexes which - together with already known structures and thermodynamic parameters - form the basis for this comprehensive study of the relationship between structure and thermodynamics. The study is performed according to highest scientific standards providing a valuable and valid basis for the deeper understanding of molecular recognition. By this, the present studie extends the existing knowledge significantly.

The authors claim also consequences for the dissociation kinetics in their conclusions and even seem to intend to investigate the binding kinetics as stated in the introduction "Here we correlate the thermodynamics and kinetics of binding with the structures ...". Unfortunately, the authors do not provide kinetic data to demonstrate, for example, the expected consequences of more complicated or retarded dissociation kinetics.

minor issues:

-l209 f   "...The observed entropies were obtained..." , the statement should be more precise
 considering the Temperature which is multiplied with binding entropy in the underlying expression.

- Fig. 6: replace Phi- by latin letter.
  The header should refer more explicitly to the mechanism of ligand dissociation.
  The arrows in the figures are misleading, because they usually indicate mesomerism between
  resonant chemical structures. If dissociation is meant, the arrow should have only one head.

-l500 f   "... Moreover, the additional hydrophobic interactions can..." instead of
   "...Moreover, the additional hydrophobic can..."
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


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

Input:
Title: CRYSTAL STRUCTURE CORRELATIONS WITH THE INTRINSIC THERMODYNAMICS OF HUMAN CARBONIC ANHYDRASE INHIBITOR BINDING
Review round: 1
Reviewer: 2

Output:
Basic reporting: The present manuscript dissert over the interaction between carbonic anhydrases isoforms and ligands, by using a large dataset of crystallographic structures of such complexes, as well as thermodynamic data of such interaction from solution techniques. The dataset is vast and the possibility of interpretation is broad, and it seems that the authors have gathered the best of their evidences and effort in order to dissect and discussed extensively their ligand binding data and active-site interaction. While I fell that such work is carefully performed and brings a large set of valuable data enlightening the field of structure-ligand interaction, in particular CA, I believe that it deserves some revision in order to improve the report in the M&M, results and discussion section as follow.
Experimental design: -	Title – Authors should consider stating in the manuscript “human Carbonic Anhidrase (hCA or just CA)” instead of solely CA. This will be clearer to the general reader, and not only to those searching specifically to CA.
-	M&M - Authors should describe in details the experimental procedure for protein expression and purification (including construct / sequence / vector) or references for the same or origin of the protein
-	M&M - Authors should describe in details the source of the ligands (synthesis, purchased, providers, or reference of the same)
-	M&M - Authors should describe in details the experimental procedure for ITC and FTSA., including (but not limited to) protein /ligand / buffer components concentration, instrumental parameters, and so on
-	M&M - Authors should describe with further details the crystallization experiments (sitting drop plate catalogue #, well volume, crystallization temperature) and preferably whether these solutions were from crystallization kits (please disclose the manufacturer) or lab-made (please disclose chemical manufacturer/provider and cat #) – this is important giving the well-known reproducibility problems associated to the crystallization technique.
-	Results – This is a ligand-focused paper. However, the protein architecture and responses over ligand binding should be addressed in the results/discussion. Even at introduction (see line 74, “The thermodynamics of ligand binding depends on many factors, including…”, and also “(Energy) gain associated with the conformational changes of ligand and protein”) no statement is done in relation to the overall protein conformation which is observed in so many protein-ligand interactions reports. A comparative analysis of the effect of ligands on overall structure might help in understanding a broader effect of ligand binding on CA´s. ProSmart (CCP4) might assist in this context, also in understanding of the whole work since several structural details are brought to the attention and without data it becomes difficult to perform a proper appraisal of such work. There is plenty of room in supporting material. Authors are encouraged to provide (as supp material) the PDB and/or a PyMOL session which could assist the reviewer and the general reader.
-	Results – line 292 “These changes may also be explained by a change in the water structure”. Please detail the meaning of “water structure”.
-	Results - How is overall hydration / ASA of the whole proteins changed according to binding?
-	Results – As stated in the manuscript (line 270 on), analysis of water based on crystallographic modeling of such molecules are limited by resolution and technique artifacts. Perhaps an analysis based on excluded volume could assist in further interpretation of such analysis.
-	Results -  “We speculate that the effects associated with the differences in the hydrophobic surface of the meta-groups have more significant impact to the binding thermodynamics than the release of water molecules from the active site by the para-benzene ring that is similar for both compounds bound to CA II.”. How do authors would consider addressing this hypothesis (themselves or others) ?
-	Results – Line 349. “All molecular pairs of similar binders indicate that energies of desolvation processes significantly change the parameters of binding thermodynamics.”. Please re-elaborate this sentence.
-	Results – Line 350. “The favorable changes of desolvation entropy after an increase of hydrophobic surface…”. Where the data supporting the statement in this sentence can be found ?
-	Results – Line 353. “The unfavorable changes of binding enthalpy in pairs can be explained by the energy…”. Where the data supporting the statement in this sentence can be found ?
-	Results – Authors claims that “The Km by both methods (FTSA and displacement ITC) agreed nearly perfectly (Zubrienė et al., 2015a). The observed entropies were obtained by subtracting the Gibbs energies obtained by FTSA from the enthalpies obtained by ITC.” Why the dG used was from FTSA and not ITC ? I´m afraid I might have missed some point or, instead, authors should address this issue accordingly.
-	Results – I feel like some statements about the magnitude and order of interaction (“strengthened by aromatic-aliphatic interactions”, “formation of additional hydrogen bonds is more likely”, “explained by increase of translational and rotational mobility
-	(freedom) of water molecules which were more ordered near the hydrophobic surfaces of
-	compounds before desolvation”, among others) relies in the speculative realm. While hypothesis might rise from experimental work, authors should rely their interpretation preferably within the data-supported context of evidences.
-	Discussion ¬– Paragraph lines 468-476. “it seems that the desolvation process is significant and may be a main reason behind the phenomenon known as enthalpy-entropy compensation.”. Authors state, with proper caution, that interpretation of water data is of limited value for a number of reasons. Should not they restrict their discussion/conclusion to this same limitation instead of using incomplete evidence from water to push some, perhaps, biased conclusion ?
-	Results – throughout the manuscript authors use the expression “pushes”. There is no action being observed here. Unless they add dynamic data here (from mol dynamic simulation, Laue diffraction, apo-holo crystallographic data, NMR, among other techniques), within the context of crystallographic evidence, they are allowed to describe the “allocation” (or any other term) of the ligand in a given place/cavity, but refrain from discussion mechanical interpretation.
-	Table 1 - Please include the Wilson B-factor for each structure in Table 1
-	Table 1 does not include all the crystal structures associated to this manuscript. Please correct accordingly.
-	Figure 1 does not include all ligands from all crystal structures reported in this manuscript. Please correct accordingly.
-	Fig 3 and 4 – These figures are really a challenge to me as a reader. They contain to many elements (colors, chemical structures, 3D-graphic representation of PX data, and a bar-chart (in c panel). Too many layers, misalignment, despite the data quality it is not good from a visual design perspective. After so much effort involved in the data collection, interpretation and writing, I´m pretty confident that a final effort can be invested in a better final Figure. Just to exemplify, Fig. 4A contains four sub-panels, three in upper line and one in lower line: the four subpanels should be leaved side-by-side for the sake of comparison. All chemical structures (Fig. 2, 3, 4) have tiny fonts, which will become not-readable on a final, reduced figure upon formatting into final paper. Please consider improving this issue.
-	Fig. 3 and 4 – again the water issue. Does it worth discuss the role of solvation water from crystallographic data ? Resolution matters. Crystallographic (ordered, thermodynamically stable ligand-bound) water molecules not the same as solvation (bulk, unordered) water.
-	Fig 5 – authors should leave the X scale (ASA) in the same scale for both “similar and dissimilar binders” (i.e., 0 to 350 A). They are encouraged to use square panels instead of long rectangles which is currently resulting in compressed Y-axis.
-	Fig 6 - authors should include in the figure a summary for their representation, independent from the legend. This would bring more prompt interpretation to the reader. Example: draw the violet circle and beside it the description “Zn”, blue rectangle next to a “sulfonamide”, and so on (mere pictorial suggestion).
-
-	Manuscript - Please carefully spell-check the whole manuscript, typos and other minor issues can be found, including (but not limited to): Pigaku instead of Rigaku,
Validity of the findings: While this manuscript bring interesting results from a large dataset (crystal, solution thermodynamics), I feel not completely confident that a large presentation of the case for hydration can be inferred from such work due to several limitations already presented here by the reviewer and the authors in their manuscript. Please consider revisiting these issues, both by limiting the scope of the discussion on hydration or by introduction of further experimental (wet bench or computer simulation) evidences supporting the body of discussion throughout the manuscript.
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: PHARMACOKINETICS AND SAFETY OF ORAL GLYBURIDE IN DOGS WITH ACUTE SPINAL CORD INJURY
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript is an experiment report rather than a scientific paper.
There are several concerns when I was reviewing this manuscript:

1. The innovation and motivation of this study are not clear.
2. The background of this study is not well introduced.
3. The reason to choose dog but not other species to conduct this study is not clear.
4. How to define the dose applied in this study is not clear.
Experimental design: 1. In the "Method", the dose used in this study is 75 mcg/kg, but in the "Conclusion" (line 323-327), the dose is 75 ng/kg.
2. In line 176, the quantification range should be from 10 - 500 ng/mL, but in  Figure 2,  the concentration level at first time point is below 10 ng/mL.
3. In table 1, both non-compartment and one-compartment models are used to calculate the PK parameters, but the authors are not telling people that which one is better, and why.
4. For the blood glucose testing, we do not know that the dogs are fasting or being given food during the study.
5. If the dogs are given food, the study design for this manuscript is not sufficient, since both food and drug may have effect on glucose level.
Validity of the findings: Conclusion is not well stated.
Additional comments: The authors should be enhanced and be more professional in the PK area.


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: PHARMACOKINETICS AND SAFETY OF ORAL GLYBURIDE IN DOGS WITH ACUTE SPINAL CORD INJURY
Review round: 1
Reviewer: 2

Output:
Basic reporting: All criteria of basic reporting are fully met.
Experimental design: All criteria of experimental design are fully met.
Validity of the findings: The data re robust and the conclusions are appropriate
Additional comments: This is a study of glyburide safety and pharmacokinetics in dogs with spinal cord injury. The manuscript is very well written with appropriate citations, the rational for the study is clear, the methods are appropriately detailed, the results are clearly presented, and the discussion lists appropriate caveats and shortcomings. I unequivocally endorse publication.

I encountered only a single difficulty that can be addressed with minor revision: in two instances, there is reference to "negative" canine plasma. It is unclear to me what the term negative implies here.


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

Input:
Title: PHARMACOKINETICS AND SAFETY OF ORAL GLYBURIDE IN DOGS WITH ACUTE SPINAL CORD INJURY
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article was written clearly in professional English, well organized and easy to follow. The authors are describing clear background of their purpose of study with proper references. However, some information and raw data might be needed.

Line 95; ‘because these other interventions and the lesion itself may alter blood drug concentrations or risk of adverse effect’.
Please mentioned whether the authors did investigate severity of injury would alter because dogs with different severity are included in Study 2 (e.g. paraparesis and paraplegia).

Line 234~&249~; Why the authors described the details of patients including age, dog bleed and body weight in Study 1 which is not included in Study 2? It is not clearly explained the omission in Study 2. Please mention why did the authors put those information in study 1 and did not in study 2.

Table 1 and supplemental table, the authors provided concentration of blood glucose level of dogs in Study 1 but not supplied those data in Study 2. Therefore, I cannot see the relation between glyburide concentration and blood glucose level, though the authors described no relation in Study 2.
Experimental design: The study design is clear; the authors investigated i) safety of oral glyburide, ii) pharmacokinetics of the drug after administration, and iii) estimate pharmacokinetic distribution in dogs with spontaneously occurred acute SCI.

However the criteria of selection of dogs and dropping out are not clearly described.
2-i) Animals; Size of dogs
Line 119; Dogs of any size were accepted for inclusion,,,,
The supplemental file (consent form final) says dogs weighing at least 5kg in ‘Eligiblility for participation’,,,, for Study2. I can see Study 1 had dog with less than 5kg and consent form for safety is not including body weight restriction. Therefore the authors may want to explane why they changed the criterion.

2-ii) Animals;
Line 123; Dogs previously diagnosed with hyperadrenocorticism or diabetes mellitus were excluded.
I think these criteria would be reasonable because those diseases might affect to the pharmacokinetics of glyburide and blood glucose level. But I can see dog 5 in study 1 had relatively high blood glucose level, and in the result section (line 263) the authors described as ‘Blood glucose concentration ranged from 50 to 324 mg/dL,,,’ in study 2. The result of 324mg/dL seems to be abnormally high concentration of blood glucose.
My questions are
-	Why Dog 5 in study 1 had relatively high blood glucose?
-	324mg/dL of blood glucose would be quite high,,,, so it would be better to explain the reason.

2-iii) Methods; Level of blood glucose
The authors set different minimum levels of blood glucose in criteria of Study 1 (50mg/dL) and Study 2 (60mg/dL). Judging from the provided reference interval of glucose level in dogs (76-119mg/dL), 50mg/dL to be low level.
The questions are;
-	Why the authors set different minimum levels of blood glucose in study 1 and 2?
-	May better to describe the concentration around 50mg/dL of glucose level would be safe in canine patients.

2-iv) Methods; 2. Pharmacokinetic study
Line 162; Bllod samples (1mL) were obtained from,,,,, after drug administration. Before each blood sample was obtained an aliquot of 5ml blood was withdrawn through the catheter and discarded.
In this study, the authors evaluated the blood samples at 13 time points during 24 hours (is this correct?). At each time point 6ml of blood was taken from dog, and in total 78ml of blood was taken from one dog during 24 hours? Is this volume of blood loss safe for small dogs (5kg of body weight)?
Validity of the findings: The data in this manuscript are not assessed previously and have valuable impacts on future studies in this field. Discussion and conclusion are focused on their primary question and supporting their results.
Additional comments: Abstract
Line 34; paraparetic
I can see dogs in study 1 showed paraplegia and dogs in study 2 showed either paraparesis or paraplegia. I am not sure whether ‘paraparetic’ would be a best word to represent these conditions.

Materials & Methods
Line 104; they receive intravenous fluid infusions
Did all dogs have same fluid infusions at same rate which might be affecting to both glucose level and concentration of glyburide?

Discussion
Line 274
I can see some dogs had exceeded level (more than 50ng/mL) of glyburide concentration at peak (Cmax). Does this concentration have any side effect to dogs? If there is an established safety range of the drug it might be better to mention.

Line277
Though I am not sure what level of blood glucose would be unsafe in dogs, around 50mg/dl seems to be unsafe for me even if dogs did not show any clinical symptom. Therefore, the authors might be better to describe why the concentration is estimated as safe in dogs.


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: PHARMACOKINETICS AND SAFETY OF ORAL GLYBURIDE IN DOGS WITH ACUTE SPINAL CORD INJURY
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript was written clearly in professional English, well organized and easy to follow. The authors are describing clear background of their purpose of study with proper references. All clear now with newly added supplemental files. Thank you for making the files.
Experimental design: The study design is clear; the authors investigated i) safety of oral glyburide, ii) pharmacokinetics of the drug after administration, and iii) estimate pharmacokinetic distribution in dogs with spontaneously occurred acute SCI. In addition the authors have added details of methods in this revised version.
Validity of the findings: The data in this manuscript are not assessed previously and have valuable impacts on future studies in this field. Discussion and conclusion are focused on their primary question and supporting their results.
Additional comments: Dear Authors
Thank you for making great revison on the manuscript according to reviewers comments. I think the manuscript has been improved with clear objectcs, methods, results and discussion. I am pleased seeing revised version of the manuscript and do not have further concern. Thanks.
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: A STRIKING NEW GENUS AND SPECIES OF CAVE-DWELLING FROG (AMPHIBIA: ANURA: MICROHYLIDAE: ASTEROPHRYINAE) FROM THAILAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English is clear and the style concise, in general the manuscript is easily read and perfectly structured. Literature review complete and up to date.

I would recommend to subdivide the discussion, for example systematics and biogeography, evolution of reproductive biology and conservation.

The illustration of the paper is outstanding.

The authors provide a fasta alignment with the new sequences. In this case, using ribosomal genes, I would suggest to include the whole alignment to facilitate replicability of analyses.
Experimental design: The description of the new species is presented in a more general systematic and biogeographic scenario, highlighting the interest of the finding. The manuscript fulfills the research standards in the field.
Validity of the findings: The manuscript presents not only the finding of a new genus and species, but provides an important contribution to the knowledge on the biogeography of the region.

The discussion is based on solid results. Even if phylogenetic relationships are not resolved in every clade, the focus is set on a well supported part of the tree.
Additional comments: The authors have done a great job in converting the description of a new genus and species into a notable contribution to the evolution, systematics and biogeography of the Microhylidae. I think the manuscript is suitable for publication and have only a few minor comments and suggestions.

I think the most important part of the discussion is the biogeographic history of the subfamily. They provide a detailed scenario. However, I think it would be strengthened if the authors test it with estimates of times of divergence of the different lineages of the group. Perhaps some of the calibration points used in previous papers could be used. Regarding phylogenetic analyses it would be interesting to check for optimal partition schemes for the data.

In the introduction, when writing about subfamilies (lines 9-10), indicate the degree of coherence between morphological and molecular classifications.

I think the introduction should be closed highlighting the implications of the new species in the biogeography and evolution of the group, as these are the main topics in the discussion.

You sometimes write the new name "Siamophryne troglodytes Gen. et sp." (example line 668) and other times "Siamophryne troglodytes Gen. et sp. nov." (example lines 687-688). Please check and unify. Is it necessary to add " Gen. et sp. nov." every time you use the name after the description?

Lines 90-92, indicate the articles of the code supporting this point.
Line 122, a bit confusing. Did you use different primers for PCR and sequencing? If so indicate which ones for which process.

Lines 128-129, confusing. Did you use all the PCR product for electrophoresis? I suggest "PCR products were visualized using 1.5% agarose electrophoresis in presence of ethidium bromide".

Line 141, analyses

Line 142, "sequences from other"

Lines 150-170, check this paragraph, it seems that some genus names are missing their authors. These part is a bit messy, perhaps it would be better to reduce it and refer to Table 2.

Line 314, "Austrochaperina is paraphyletic...". Is it possible that they are misidentifications?

Line 320-321, I think the sentence should be rewritten: p-distances among and within gene fragments or among and within genera?

Line 328-329, "gene of the new species (Table 3)".

Line 348, minimum size should be 19 mm, not 20.

Line 374-375, " Asterophryinae genera which have symphignathine".

Line 423-424, "expanded and broad cultiform process of parasphenoid in..."

Line 457, I think numbers are provided for only 9 specimens.

Line 547, " are larger than males".

Line 793-794, just before this sentence you stated that nothing is known about the reproductive biology of Gastrophrynoides, so it sounds a bit speculative.

In table 2 and its caption, would not it be possible to use the full name of the new species? Same for table 3.

In table 3 caption it says: " The mean uncorrected p-distances for the ingroup are shown in the diagonal and shaded with grey.". Are these not distances within some genera? Check.


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

Input:
Title: A STRIKING NEW GENUS AND SPECIES OF CAVE-DWELLING FROG (AMPHIBIA: ANURA: MICROHYLIDAE: ASTEROPHRYINAE) FROM THAILAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper describes a very interesting new species of microhylid frog from Thailand, Generally the quality of the research and inferences made based on the data seem highly justified and reasonable.

This said, there is perhaps a missing opportunity to link in with other similar finds of high diversity, restricted or relict diversity in karst habitat, e.g see a number of papers on herps:
Glaw et al. 2006 Zootaxa, 1334, 27-43 Tsingymantis
Kohler et al. 2010 Journal of Zoology 282, 21-38.
Oliver et al. 2017 Diversity and Distributions, 23, 53-66
Grismer et al. 2017 eg latest in Biol. J Linn Soc (and many others)
And perhaps Clements et al. 2006, Bioscience 53,733-746 - for a more general ref

Likewise in lizards a least there is literature on morphological adaptations that characterise cave dwelling taxa, does this new species so any adaptitions typical of cave vertebrates – reduced pigmentations, longer limbs etc?

The discussion of biogeography seems a little long, it already pretty well established that Asterophryne has Asian origins, this new find supports and strengthens this interpretation, but less discussion of the alternative hypothesis (Gondwanan origins) is needed.

Results – suggest delete all discussion of relationships within the Melanesian Asterophyrninae (lines 302-314)– it is not relevant to this paper, the data upon which this discussion is based is not new, and more recent studies – e.g Rivera et al 2017 have better data for this job anyway
Experimental design: All fine.
Validity of the findings: Data, inference and overall findings seem robust.

Conclusions are all reasonable, however as noted above there are several paragraphs focused on relationships within the New Guinea Asterophyrnes that are recapitulations of data presented elsewhere, the data avaliable (12S and 16S) does a poor job, and better phylogenies have already been published. These sections should be cut.

As noted above, most speculation is reasonable, but I would also suggest that is a missed oppurtunity here to link into and emerging literature on a global story of karst herpetofaunas (endemism, relictualism and specialisation).
Additional comments: Minor comments

Line 17 – Gondwanan landmass = Gondwana?

Lines 35-45 While the placement of Gastrophrynoides in association with the Melanesian taxa is probably right, its placement within that clade is likely wrong (based as it was on small amout of heavily saturated mtDNA), and as alluded to later, so why report this result in so much detail?

Lines 49-51 – saying same thing twice, but in different ways

Line 72 – personal bugbear, but, please use decimal co-ordinates – the more people use these the easier processing this data will be

Line 118 – could add a few more taxonomic studies – e.g.
Hoskin 2004 Australian Journal of Zoology, 52, 237-269
Oliver et al. 2017 PeerJ 5,e3077
Conversely Blackburn paper was not focused on Asterophryninae

Line 150-162 – large bracketed section is very awkward, would easirer to read if broken up into several sentences

Line 262 – sequence alignment, how was this done, and presumably there were unalignable regions that were chopped out?
Line 374 – ‘thereunder’ = “thereby?”

Line 388 – s. lato = sensu lato (spell out fully, or abbreviate full)

Line 396 “can be distinguished from the new genus by reduced clavicles (vs. clavicles present)” – this does not make sense

Line 468 – ‘limbs very long’ – would be great to add some relative proportions measures to expand on this

Line 479-480 – this is not a trait that can be scored from specimens, so I would suggest removing

Line 486 – change “’,” to “;”

Line 496 – “more than two times shorter” = “less than half length”?

Line 537 – what colour is the ‘dark mottling”?

Line 668 – Comparisons (or somewhere) – if possible it would be nice to include an image of Gastrophrynoides to emphasis how different these taxa are, even though they are sister species. They provide another example of the ecological plasticity of microhylids – e.g Blackburn kaloula etc.

Line 687 – are there many similar caves nearby? Likewise is there exposed karst habitats that they could live amongst?

Line 752-776 – this paragraph restates other peoples results and can be deleted

Line 777-778  - delete first sentence – no one has ever found a microhylid tapdole in New guinea – they are all almost certainly all direct developers

Final para of discussion – see above major comments about a missing opportunity to link in with a broader literature on cave endemism in herps, and the conservation importance of caves

Line 821 – change to ‘only two’


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

Input:
Title: A STRIKING NEW GENUS AND SPECIES OF CAVE-DWELLING FROG (AMPHIBIA: ANURA: MICROHYLIDAE: ASTEROPHRYINAE) FROM THAILAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article is well written and results are clearly presented with the help of tables and figures that have been carefully prepared.
Experimental design: The article reports the finding of a highly divergent lineage of microhylid frog, which carries important biogeographic implications. The authors show the distinctiveness of this new lineage with mitochondrial DNA sequences, morphological (internal and external), and ecological data. The phylogenetic analyses are correctly conducted and presented and the description of the osteology is very comprehensive and informative.
Validity of the findings: While only mtDNA sequences are used to reconstruct phylogenetic affinities of the new taxon, the results clearly show its position as an early branching member of the Asterophryinae. The biogeographical implications of the results of these analyses are carefully discussed in light of previous hypotheses.
Additional comments: I congratulate the authors on a very thorough study on a very interesting finding. I only have a few minor suggestions for corrections in the text (see below):

Line 18: Madagascar

Line 146: in fact, it's 56 microhylids plus the outgroup

Line 164: Melanobatrachus

Line 181: Confidence in tree topology was evaluated/assessed rather than tested

LIne 223: add abbreviation (2-5 TOEL)

Line 230: ...those of Savage

Line 290: Asterophryinae consists of two major...

Line 293: this should read east of the Wallace line, not west

Line 317: Gastrophrynoides

Line 328: this should be 12S rRNA-16S rRNA

Line 337: inhabits

Line 412: lacks these structures

Lines 421 and 423: cultriform process

Line 534: refer to Fig. 5

Line 614: this would be Table 2, not Table 1

Line 717: Gastrophrynoides

Lines 727, 730, 731: rather than "had", I'd either use "would have" or delete "had" altogether in the three cases

Line 748: replace "initial origination and cladogenesis" by "origin and early cladogenesis"

Line 768: Liophryne

Line 776: replace "better" for "robust"

Line 785: replace "as well" for "either"

Line 814: relict

Line 820: Gastrophrynoides (check spelling throughout)

Figure 2 legend: GB accession numbers are listed in Table 2, not Table 1. Also, either explain that the A in the tree corresponds to the Asterophryinae node or delete from the figure

Figure 6 legend: check spelling for phalanges (mis-spelled twice). Also, although it is referred to in the legend, I could not find the prehallux in the figure

Figure 7 legend: although it is referred to in the legend, I could not find the vomer in the figure

Table 1: there are six primers listed to amplify the 12S fragment, perhaps you could clarify in the text whether these correspond to nested PCRs or some are sequencing primers. If this information is presented in Nguyen et al (in press) forget about this


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

Input:
Title: ASSESSMENT OF THREE RISK EVALUATION SYSTEMS FOR PATIENTS AGED ≥70 IN EAST CHINA: PERFORMANCE OF SINOSCORE, EUROSCORE II AND THE STS RISK EVALUATION SYSTEM
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The author tried to assess and compare the predictability of three risk evaluation systems  in patients aged≥70 , and who underwent coronary artery bypass grafting (CABG) in East China. Then they concluded"good discrimination and calibration in the entire cohort. In the elderly group, SinoSCORE achieved slightly better predictive efficiency than STS risk evaluation system". The manuscript can be improved significantly by performing the following:
1.	Though the cohorts are large, due to the single center and single ethnicity (East China) study make it impossible to make such a strong conclusion and should explicit for limitation in the discussion section.
2.	Please clarify your bias when select the population prior to your conclusion.
3.	L81-82, please state why and how you classified the subgroup? Why aged 70 not others?
Your paper may be conditionally accepted after “major revision” and language polishment.


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: ASSESSMENT OF THREE RISK EVALUATION SYSTEMS FOR PATIENTS AGED ≥70 IN EAST CHINA: PERFORMANCE OF SINOSCORE, EUROSCORE II AND THE STS RISK EVALUATION SYSTEM
Review round: 1
Reviewer: 2

Output:
Basic reporting: While the text was generally clear in its intention, there were a number of typos or instances of awkward wording. Careful proof-reading and editing for style are both necessary. Some suggestions for the authors to consider in this regard are listed below under “General comments”.

Important details are sometimes omitted from the manuscript, leaving the reader to guess at what has been done. In other cases, details are provided later than I would expect; for example, the abstract’s objective and method sections both fail to mention that mortality was the endpoint of interest (I appreciate that this is almost inevitable as the outcome but the reader should know this for certain rather than expecting it to be the case). AUCs are presented in the abstract results without indication that this is what they are. The elderly group is discussed in the abstract without being defined.

Some of the details of the statistical analyses were only entirely clear after I had looked at the data, which was in a pleasibly sensible format and I thank the authors for this. These details include the reported mortality statistics which appear to be mean scores from the risk equation systems along with standard deviations (which are potentially problematic given the substantial skew in the distribution) and 95% CIs for these means (again, skew being an issue that needs to be considered).

The introduction needs to be considerably expanded to establish the context of the study and to motivate the research (experimental design). I would suggest a discussion of each of the scoring systems for the unfamiliar reader (otherwise, it is not until the discussion when the numbers of component items and some examples of these items are discussed that a reader not already familiar with the instruments is given any clue about their structure or complexity).

Also, references are made to literature looking at homogeneous patient groups and a lack of literature looking at different ages (lines 59-60), but discussion of this existing literature is lacking.
Experimental design: The knowledge gap that this manuscript aims to address should become clearer with the extra context recommended above.

I am assuming that all patients had been discharged when data was extracted but a statement to this effect (if it is not already present) would be reassuring, along with information about length of stay post-op.

I am concerned about the clearly skewed variables being described using means and standard deviations in all cases. While the arithmetic mean can be appropriate for mortality (and could also be appropriate for length of stay if this was added), the standard deviations are greatly inflated by the substantial skew in risk scores, for example. For age, perhaps median and IQR would be more useful if these are skewed as I suspect?

Related to this, no description of model diagnostics for the t-tests or the basis for deciding between Fisher’s Test and the Chi-squared tests is given (lines 93-94). Mann-Whitney-Wilcoxon would be an alternative should the assumptions of the t-test not be satisfied based on raw or transformed data.

Hosmer-Lemeshow is a test of model misfit (lines 99-100) and so a non-significant test does not indicate a well-calibrated model, merely one where there is a lack of sufficient evidence for miscalibration. The wording in the text should reflect this interpretation. Also, marginal test results (such as those on lines 129) would still concern me when there are so few events and so limited power in the test to detect departures. Note also that the test does not require deciles to be used (line 99), although this is a common choice.
Validity of the findings: The abstract states that the objectives include comparing the three systems, but no formal comparisons appear to be included beyond the inclusion of what might be a comparison between the three AUCs in the figures. Given the apparent lack of statistically significant differences in AUC between the three models (based on these p-values), the interpretation that SinoSCORE is superior in any sense beyond a simple numerical comparison of values needs to be softened (in the results and in the discussion). While the empirical estimates for AUC for SinoSCORE are indeed higher than the other systems in two of the three models, and so that might make it a reasonable choice for an organisation not already using any risk evaluation system when looking at older patients or patients in general, the lack of evidence for differences provides no strong basis for an organisation already using EuroSCORE II or STS to switch to using SinoSCORE, for example. A similar point applies to the interpretation of the DCA results (lines 151-153).

Given that all three systems underestimate mortality by what I would consider to be an unacceptable degree, I think that this suggests overall problems with the modelling of risk and I have to disagree with the statements that “good” or “positive” calibration was achieved. Given the substantial misfit from the Hosmer-Lemeshow test, I would suggest investigating the inclusion of a quadratic term, for example. Another possible cause of this misfit could be an omitted main effect, or after the addition of such, possibly one or more interaction terms. However, the sample sizes of events is small and should constrain the complexity of the statistical models, especially the age stratified models (e.g., Peduzzi, et al. using their 10 EPV recommendation).

The authors correctly identify an issue with missing data where these cases will not involve mortality. Multiple imputation would be a useful way of incorporating this data back into the analysis, correcting for this bias. This could be done as the primary analysis with the complete data as a secondary analysis, or vice versa.

The stratification by age is interesting. Given the two hospitals providing data, I would like to see this as another stratification variable. This would also help to address the issue of clustering within hospitals. If generalisation beyond these hospitals is not required, and the authors did not want to stratify by hospital, a fixed effect for the hospital could be added to logistic regression models to allow for evaluating the scores (note the limits of model complexity discussed above though). I wonder if some of the other patient characteristics could be added to age (and perhaps hospital) in this way?

The use of the same patients who were used to develop SinoSCORE (lines 188-189) is a problem as this makes the estimated performance using this instrument overly optimistic. The analysis should exclude those patients used to develop this instrument to avoid this problem and thereby provide less biased estimates of the model’s performance.
Additional comments: This manuscript examined three risk evaluation scoring systems (EuroSCORE II, STS, and SinoSCORE) as predictors of mortality from 1987 CABG patients from two hospitals in China (51 deaths occurred). The three scores are considered overall and using two age strata (<70 years, 70+ years). Note that these categories contradict the manuscript title which refers to those aged 70 and above. There is considerable misfit from each risk prediction model and it appears that no statistically significant differences were found between the three scores, although SinoSCORE was empirically better fitting according to AUC for two of the three models.

The manuscript seems well-motivated from my (a statistician's rather than a clinician’s) perspective, but the issues of poor model fit (the calibration component) and over-interpretation/lack of discussion of what appears to be no evidence of superiority of any score over the others means that the data has not been used to its full effect.

A non-exhaustive list of suggestions for improving the wording includes (with new words and/or suggested replacements in upper case):

1.	“predictability” in the abstract (line 29) might be better as “predictive ability”.
2.	I would delete the first “statistics” from the abstract’s methods, or make both singular (line 35).
3.	Abstract: “1.51% in THE younger” (line 40), “best predictive value in THE elderly group” (line 41), “while ALL three risk evaluation systems had” (line 42).
4.	Use a more precise description than “They” (line 52).
5.	“systemS play an” (line 53).
6.	“THE Society” (line 56).
7.	“operations AND HAVE rarely evaluated their ability” (line 60).
8.	“were INVITED TO PARTICIPATE in the study” (line 68).
9.	The nature of the missing information (line 71) needs to be clarified.
10.	Delete “have” (line 71).
11.	“incomplete data” (line 72).
12.	“average” is presumably “mean” (line 74) and while plus/minus notation is commonly used to indicate standard deviations, this should be defined for the first such use.
13.	The number of decimal places (e.g. line 74) seems excessive to me.
14.	A flowchart showing the information about the patients invited and eventually analysed (lines 66-79) would be useful.
15.	Presumably, the level of significance is two-sided? (line 94)
16.	“acceptable” rather than “available”? (line 103)
17.	“OF A 45º line, wheRE” (line 106).
18.	For R packages, the actual name should be used (line 113) and I’d suggest this be a reference with a link to CRAN or elsewhere.
19.	“MEAN age” (line 117).
20.	Note that while SPSS reports p-values as “0.000” in some cases, the p-value is of course not actually zero and I would recommend reporting such small p-values as “p<0.001” instead.
21.	The number of diseased vessels (line 121) is presumably referring to the mean number of there? Given the discrete nature of this variable, I would have suggested Mann-Whitney-Wilcoxon instead. The number of grafts is a simialr case.
22.	Line 125 seems to be present mean mortality percentage estimates rather than rates.
23.	AUCs should be accompanied by CIs (e.g. lines 130-131). Note these appear to be included in the figures but are not labelled as such there.
24.	“in THE elderly group IS shown in” (line 141).
25.	“BOTH showed” (line 146).
26.	“systemS” (line 148).
27.	“for THE younger” (line 156).
28.	“in THE younger group IS shown” (line 158).
29.	“THE younger” (line 162).
30.	“THE EuroSCORE” (line 170).
31.	“world, HAVE BEEN well” (line 171).
32.	Lines 181-182 are presumably about confidence in the use of the system?
33.	Should “epidemiology” be something about patient characteristics, health behaviours, disease progression or similar?
34.	I would move the Discussion points about characteristics of the SinoSCORE patients to the results (lines 189-193).
35.	“performance” rather than “power”? (line 196)
36.	“ALL had” (line 197).
37.	“substantially” rather than “sensibly”? (line 198)
38.	“THE elderly” (line 199).
39.	“published RECENTLY, SincoSCORE” (line 228).
40.	“no compromise” needs rewording (line 230).
41.	For the figures, there appear to be 95% CIs in parentheses and these should be labelled as should the p-value (which I assume to be comparing all three AUCs).
42.	For the tables, I would suggest “n” rather than “N” for the sample sizes.
43.	Table 2 suggests the p-values are from Cox’s proportional hazards models, which doesn’t match the methods.
44.	For surgery status, I am assuming this was a three-level variable and so compared with one and not three tests. The overall p-value would be more appropriate than the three pairwise comparisons.
45.	Table 3’s headings could be clearer so the reader does not need to read the abbreviations.


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: ASSESSMENT OF THREE RISK EVALUATION SYSTEMS FOR PATIENTS AGED ≥70 IN EAST CHINA: PERFORMANCE OF SINOSCORE, EUROSCORE II AND THE STS RISK EVALUATION SYSTEM
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This revised MS could be considered to be accepted after the chages made.