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: CONGRUENCE BETWEEN MORPHOLOGY-BASED SPECIES AND BARCODE INDEX NUMBERS (BINS) IN NEOTROPICAL EUMAEINI (LYCAENIDAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: The manuscript is overall clean and clearly written, but some light editing is needed throughout, e.g., "These phenomena seem to be particularly common in high Andean genera such as Rhamma and have important effects on species identification based on genetic sequence analysis" - compare with the original sentence.

I suggest that some sections of the manuscript could be improved by clarifying some issues and providing greater background information and references to relevant literature, especially more recent studies. The Introduction could clarify two of the main goals of developing DNA barcode libraries and go on to say why both of these are relevant to the study group. More information could be presented about the background of the study group. And, throughout the manuscript, there are a number of studies, especially published in the last 10-15 years, that would be relevant to cite in relation to Neotropical butterfly biogeography, e.g. on various groups of Melitaeini, Ithomiini, Adelpha, Heliconius, Pronophilina, Papilionidae, Riodinidae.

The authors are to be commended on their thorough approach to making all relevant data publicly available through BOLD.
Experimental design: Overall, the methods used and the data used are very well described and justified, with the exception perhaps of a little more detail about the parameters in RESL analysis, and why alternative delimitation approaches were not used could be better justified.

As mentioned above, the knowledge gap being filled (existing library of eumaeine sequences and understanding of eumaeine taxonomy) could benefit from greater clarification in the introduction. I also suggest that the authors consider conducting RESL analyses for subsets of their data, e.g. split into 'lowland' and 'montane', to see whether methodological issues such as trying to fit parameters to a highly variable dataset are reducing the effectiveness of the approach in montane groups. Furthermore, an analysis of the extent to which failures of the analysis to discriminate species result from low divergence between monophyletic groups vs parar/polyphyly would be very useful in directing subsequent discussions about the causes of these analytical failures. Finally, and throughout the manuscript, there should be a constant awareness about the importance of geography in explaining barcode divergence and affecting the apparent success or failure of species delimitation, both in this and other studies.
Validity of the findings: This is a massive and very significant dataset for a highly complex and understudied group, yet I think the authors could still better emphasize and quantify this contribution.

The conclusions are largely sound, except a bit qualitative, and there is a strong emphasis on discussing recent diversification and incomplete lineage sorting, whereas it is not clear how many cases of failed delimitation are due to these issues. The discussions of Andean butterfly biogeography are sometimes a bit generalized and lacking in references to recent publications, as mentioned above.
Additional comments: Overall, this is a really remarkable study which will provides a huge dataset of great value to Neotropical butterfly researchers. I made some more specific comments below.

Background
I am not sure infraspecific variation in Lycaenidae can be considered 'great', in comparison with many Nymphalidae, for example. The significant point is that geographic infraspecific variation can appear more substantial than variation between species, which is often very slight.

Methods
It seems inaccurate to say that it is the BIN system that delimitates MOTUs, rather MOTUs are delimitated by the RESL algorithm and each is labeled with a Barcode Index Number (BIN).

L 57: 'enumeration' is a bit vague, DNA barcodes can provide an estimate of how many species are in a sample without needing to put names on any of them (and hence without needing a reference library).

L 73: Elias et al 2007 is not a very appropriate reference here, instead it would be better placed on line 76 as a counter-example of the usefulness of barcodes. Better examples from Lepidoptera, in which descriptions of taxa were actually facilitated by DNA barcodes, can be found, such as Dias et al. (2019, "DNA barcodes uncover hidden taxonomic diversity behind the variable wing patterns in the Neotropical butterfly genus Zaretis (Lepidoptera: Nymphalidae: Charaxinae)") or Espinoza et al. (2017, "17 new species hiding in 10 long-named gaudy tropical moths (Lepidoptera: Erebidae, Arctiinae").

L 80: "The usefulness of DNA barcodes for describing biodiversity patterns depends on a strong correlation between morphologically and genetically delimited species" – again, this is a bit unclear. DNA barcodes have two common and distinct applications – 1) as a supplementary set of characters alongside morphology, ecology, other DNA markers, or anything else that can be used, to help delimit species and build a classification; 2) as a tool for identifying unknown samples by matching them to a library of identified sequences. These two points are mentioned in L 115-116, but I think they could be made more clearly at the start of the Introduction, to lead on to which of these aspects, or both (as is likely), will benefit from this study and dataset.

L 94-96 need a bit more elaboration for those not familiar with the group; what is meant, for example, by 'hard sampling' or 'restricted geographic distribution'? A few sentences about their biology might also be nice, e.g. behavior, hostplants, longevity, broad geographic patterns of community diversity etc. Furthermore, a couple of sentences about the state of Neotropical eumaeine taxonomy would be useful too – how many undescribed species are known, how frequently are new undescribed species uncovered, what are the principal morphological characters that have been used to build the current classification, are there large genera so in need of complete revision that their current classification is really rather tentative?

L 109-110: Or, in many cases, the decision is to gather further information, such as additional genetic markers or add samples to fill in geographic variation, to help resolve the situation.

L 117-118 (and L 167-169): this hypothesis presumably refers back to the empirical observation mentioned in L 102-103, but if this is an important goal of the paper it could merit a bit more discussion, e.g. theoretically why might high elevation species be less easy to delimit/identify? Lower gene flow because of linear ranges and fragmented populations, and therefore greater infraspecific geographic variation in barcodes? More recent speciation because these habitats are geologically younger? Habitat filtering which tends to lead to less phylogenetically diverse communities at higher elevations (e.g., Chazot et al. 2014, "Mutualistic Mimicry and Filtering by Altitude Shape the Structure of Andean Butterfly Communities")?

L 126-127: would seem more accurate to me here to say identification of samples, rather than species.

L 136: 541 species identified a priori based on the existing classification, presumably?

L 145: Whenever I mention 'Neighbor-joining' it brings down the wrath of most reviewers, so some justification as to why you used this method to construct trees (vs a ML or Bayesian approach, for example) would be good. What nucleotide substitution models were used, and what about other kinds of settings (e.g., dealing with missing data)?

L 155-156: what is beneficial about a 'deterministic' method, and what does that mean anyway? Also, being faster is not necessarily a reason for using a method, what other reasons support use of the RESL algorithm over other species delimitation approaches, e.g. GMYC, PTP?

L 185: not clear why the 'congruence analysis' contains only 1597 of the 1839 x >500 bp sequences obtained.

L 192-195: This is unclear, how can species that 'cannot be discriminated by DNA barcodes' have 'diagnostic barcodes'? Furthermore, what is this 'taxonomic decision', presumably you did additional genitalic dissections or analysis of other characters and found consistent differences between samples that coincided with DNA barcode differences, supporting the need to re-evaluate species limits? This retroactive revision should be described in the methods.

L 198: I may have missed this in the Methods, but what does this percentage indicate? How exactly was it determined whether an individual species 'had' a barcode gap? Ah, I see it is described rather well in L 273-278, I suggest moving this section to the Methods.

L 209: I'd suggest opening the Discussion with a summary statement about the main outcomes of the study, e.g. 'In this study we obtained X new DNA barcodes for Y species of Neotropical Eumaeini, increasing by Z% the number of species for which there are barcodes now available in this group. We found such divergence corresponding to etc etc.' Then move on to the details.

L 214-215: this is a bit of a woolly statement, who is to determine whether a particular taxonomist is 'competent'?

L 219: 'particularly', rather than 'mainly'

L 222-225: some references to support these assertions would also be useful. Furthermore, some discussion about the critical aspect of geographic distance between samples would be useful. A 2% divergence between samples from the same site is likely significant, but a 2% divergence between samples from Mexico and SE Brazil is almost meaningless.

L 228: sympatric morphospecies, or allopatric? If allopatric, their 'species' status is subjective.

L 234-236: Would it be worth exploring analyses of smaller subsets of the data, e.g. if you repeated the analysis for just 'montane' genera, would different parameters be selected and a better agreement between MOTUs and traditional species be found?

L 237: "A relatively small number of cases of BIN sharing represent paraphyly or polyphyly that could result from incomplete lineage sorting…", some introductory statement is needed to tie this to the previous paragraph.

L 250-253: Also, critically, as mentioned above, intraspecific divergence can reflect geographic separation, e.g. 6% divergence in Amiga (formerly Chloreuptychia) arnaca between eastern and western Ecuador.

L 256-257: Presumably, you mean conspecific sequences from distant localities were more likely to be divergent, but this sentence could also mean that DNA divergence between samples of species 1 from sites A and B is greater than in species 2 also occurring in sites A and B, when overall species 1 has a broader geographic distribution than 2. So, some clarification is needed.

L 259-260: again, do these BINs correspond to geographic clusters, or do they occur in sympatry?

L 285-286: "The Andes rose much later (about 25 MY ago)" is a bit of an oversimplification, with the southern Andes being much older than the northern Andes, and the 'age' of the mountains throughout being strongly dependent on the elevation above sea level that you are discussing. Furthermore, the statement "and were populated by butterflies originating in the eastern parts of the continent'" is also a large over-generalization.

L 287: Andean butterflies? Butterflies in general? Neither statement is generally correct though, since peak diversity in the eastern tropical Andes is usually around 700 m, and in the western Andes a bit higher, and obviously it differs greatly depending on the group, I imagine Riodinidae peak at around 400-500 m for example, in the eastern Andes. More informative here would be a statement about where Lycaenidae diversity peaks.

L 292: 'intrincated' should be 'intricate', plus inclusion of some references to support this statement would be good, e.g. papers on Adelpha, Heliconius, which include broad maps of Neotropical butterfly diversity.

L 293: 'endemic to the valley system' – not really clear what you mean by this. Few species are endemic to individual valleys.

L 295-296: again, some references would be useful, e.g. papers on pronophilines.

L 296-303: more references needed, including more recent references.

L 304-305, reword to clarify and avoid repetition.

L 314-317: but these phenomena explain paraphyletic and polyphyletic species, whereas some monophyletic species with low separation may have been lumped as an artifact of the broad scale of the analysis. It would be nice to quantify how many of the problem BIN sharing was due to these two different phenomena.

L 318-319: very important to make sure you are comparing similar scenarios. For example, a study of barcodes in a single site is more likely to find barcode gaps than one based on the same taxa across a broader geographic area. A study of barcodes in one less diverse site is more likely to find gaps than one in a more diverse site (where sister species are more likely to co-occur). Finally, a study of barcodes in genus may also be more likely to find barcode gaps if sampling within each species is geographically very limited.

L 322-325: this is an interesting hypothesis that could be better supported if it was known how many cases of BIN sharing resulted from hybridization vs low divergence between clades (i.e., biological vs methodological issues, as mentioned above?.


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

Input:
Title: CONGRUENCE BETWEEN MORPHOLOGY-BASED SPECIES AND BARCODE INDEX NUMBERS (BINS) IN NEOTROPICAL EUMAEINI (LYCAENIDAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is overall clean and clear. However there are spelling errors and in the manuscript the references are out of chronological order. All of them are marked in yellow in the attached file.
More importantly, in lines 115-116 they say 'A specific objective of this paper is to test the hypothesis that the ability of DNA barcode BINs to discriminate morphologically delimited species decreases in high elevation lineages', but this it is an objective, not an hypothesis. The hypothesis is a proposed explanation made on the basis of some evidence, as starting point for further investigation, so I recommend that the authors make the hypothesis and explain why the DNA barcode decreases its ability to discriminate in high elevation linages.

The results are relevant and deserve to be published after minor corrections and clarifications.
Experimental design: The data and findings of this paper are valuable. This group of butterflies are one of the most complicated for several reasons, and the authors explore the effectiveness of this barcode tool as a first approach to the problem.

They added some statistical test; however, perhaps they should prefer other tests to find out whether or not the variation is significant between their comparisions. I suggest ANOVA or Kruskal Wallis test
Validity of the findings: This contribution is important. The massive data that the authors generated will be important for future research. This is a first approximation to the usefulness of the barcode as a tool to identify species that are first morphologically discriminated. The conclusions are appropriate. Also, this can still improve on the statistical part as suggested above.
Additional comments: It is an important contribution for the taxonomic group and for the high diversity area.

Even so, there are still errors that need to be corrected and the statistical part can still be improved with the suggested tests. The same with the hypothesis in lines 115 and 116, since they are not an explanation of the facts that you observed. All of them are marked in yellow in the attached file.


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: CONGRUENCE BETWEEN MORPHOLOGY-BASED SPECIES AND BARCODE INDEX NUMBERS (BINS) IN NEOTROPICAL EUMAEINI (LYCAENIDAE)
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed most of my comments from the previous submission, I just added a few new comments or requests for clarification in the manuscript file.
Experimental design: More detail was added about the RESL analysis, but still it is difficult to understand what a priori settings are needed for this analysis, and since the paper refers to these settings in a couple of places, I think a more detailed description would be useful of how the analysis works and what a priori settings decisions need to be made.
Validity of the findings: The authors have added some additional references and discussion about Andean butterfly biogeography, and I still think that the conclusions are largely sound, if a bit qualitative, but it is the authors' decision whether or not to delve further into their findings in this paper, or leave it for a future study. I am happy with either one.
Additional comments: Overall, this is a really remarkable study which will provides a huge dataset of great value to Neotropical butterfly researchers, and I think the revised version addresses most of my concerns/queries, except for notes in the attached manuscript.


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

Input:
Title: CONGRUENCE BETWEEN MORPHOLOGY-BASED SPECIES AND BARCODE INDEX NUMBERS (BINS) IN NEOTROPICAL EUMAEINI (LYCAENIDAE)
Review round: 3
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: The authors have addressed my comments from the previous review and I feel the manuscript should be accepted as it is.


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: PAIN AND SATISFACTION: THE CASE OF ISOLATED COVID-19 PATIENTS OF PAKISTAN
Review round: 1
Reviewer: 1

Output:
Basic reporting: - The paper needs proofreading and grammatical editing as grammar issues have been prevalent throughout the manuscript. I would recommend rewriting various sentences and subject/verb agreement issues are widely present all over the text.
- As for the article, it is well structured in general.
- The abstract needs to include numbers.
- Old references in the discussion need to be updated.
- Add an implications section
Experimental design: - The research design is not well described.
- Chronbach's alpha for the questionnaire was not assessed or mentioned.
- Statistical analysis was undertaken as appropriate.
- Add response rate
- Add the number of ethical approval
- Add sampling methodology and related reference
Validity of the findings: - Add frequencies to table 1
- Results are adequate. However, more tests are advised to enrich the findings and giving a higher impact to the paper.
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: PAIN AND SATISFACTION: THE CASE OF ISOLATED COVID-19 PATIENTS OF PAKISTAN
Review round: 1
Reviewer: 2

Output:
Basic reporting: The study is timely and deserves merit.
Experimental design: The methodology employed is valid but few clarifications are required. (see comments below).

Also, do include a model that shows the hypotheses tested.
Validity of the findings: Findings are well presented.
Additional comments: Pain and satisfaction: the case of isolated COVID-19 patients of Pakistan
Below are comments to improve the manuscript
Include tool used for analysis in the abstract (STATA and Smart PLS-3).
Elaborate more on the issues and need for the research in the introduction section.
Also, provide research questions to be examined in the study in the introduction section.
Add a paragraph of the summary of the sections in the last sentence in the introduction section.
Include literature review section (related works) section as section 2, that reviews prior COVID-19 studies such as
•	Amol Soin, M. D., Srinivas Vuppala, M. D., Gregory Surfield, M. D., Ricardo Buenaventura, M. D., Mark Malinowski, D. O., Rajaratnam, A., ... & Chandoke, A. (2020). Ohio response to COVID-19 and its impact on interventional pain management practices. Pain physician, 23, S439-S447.
•	Anthony Jnr, B. (2021). Implications of telehealth and digital care solutions during COVID-19 pandemic: a qualitative literature review. Informatics for Health and Social Care, 46(1), 68-83.
•	Song, X. J., Xiong, D. L., Wang, Z. Y., Yang, D., Zhou, L., & Li, R. C. (2020). Pain management during the COVID-19 pandemic in China: Lessons learned. Pain Medicine, 21(7), 1319-1323.
•	Anthony Jnr, B. (2021). Integrating telemedicine to support digital health care for the management of COVID-19 pandemic. International Journal of Healthcare Management, 1-10.
Also, do include a model that shows the hypotheses tested.
The methodology is well written but provide more information on the sampling technique employed.
Provide full meaning of STATA and Smart PLS-3 and why was STATA and Smart PLS-3 employed and not SPSS?
There are results on factors affecting pain level. How was the factors derived and do present these factors in a less vague approach as this is one of the contribution of the study?
Include standard deviation for descriptive analysis.
How was reliability and validity measured?
Include a section on implications of the study.
Provide discussion on the limitations and future works in the conclusion section.


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: PAIN AND SATISFACTION: THE CASE OF ISOLATED COVID-19 PATIENTS OF PAKISTAN
Review round: 3
Reviewer: 1

Output:
Basic reporting: Well presented as the manusrript was revised.
Experimental design: Acceptable
Validity of the findings: Can be improved as some results are missing.
Additional comments: Pain and satisfaction: the case of isolated COVID-19 patients of Pakistan
Below are comments to improve the revised version of the manuscript

Also, provide research questions to be examined in the study in the introduction section. The research question should be in bullet point

Include literature review section (related works) section as section 2, that reviews prior COVID-19 studies such as
• Amol Soin, M. D., Srinivas Vuppala, M. D., Gregory Surfield, M. D., Ricardo Buenaventura, M. D., Mark Malinowski, D. O., Rajaratnam, A., ... & Chandoke, A. (2020). Ohio response to COVID-19 and its impact on interventional pain management practices. Pain physician, 23, S439-S447.
• Anthony Jnr, B. (2021). Implications of telehealth and digital care solutions during COVID-19 pandemic: a qualitative literature review. Informatics for Health and Social Care, 46(1), 68-83.
• Song, X. J., Xiong, D. L., Wang, Z. Y., Yang, D., Zhou, L., & Li, R. C. (2020). Pain management during the COVID-19 pandemic in China: Lessons learned. Pain Medicine, 21(7), 1319-1323.
• Anthony Jnr, B. (2021). Integrating telemedicine to support digital health care for the management of COVID-19 pandemic. International Journal of Healthcare Management, 1-10.
Also, do include a model that shows the hypotheses tested.
How was Normality  tested in the study if SPSS was not used.
Provide full meaning of SPSS
There are results on factors affecting pain level. How was the factors derived and do present these factors in a less vague approach as this is one of the contribution of the study?
Include standard deviation for descriptive analysis.
How was reliability and validity measured?
The section implications of the study should be placed before the conclusion section


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: PAIN AND SATISFACTION: THE CASE OF ISOLATED COVID-19 PATIENTS OF PAKISTAN
Review round: 4
Reviewer: 1

Output:
Basic reporting: The author has incorporated the needed changes according to comments.
Experimental design: The author has incorporated the needed changes according to comments.
Validity of the findings: The author has incorporated the needed changes according to comments.
Additional comments: Dear author,
Thank you for your 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: PAIN AND SATISFACTION: THE CASE OF ISOLATED COVID-19 PATIENTS OF PAKISTAN
Review round: 4
Reviewer: 2

Output:
Basic reporting: The paper is grounded on the literature. Also, the research questions employed has been provided.
Experimental design: The methodolofy employed is well presented. Besides, the sampling method has been presented.
Validity of the findings: Data  analysis was performed with STATA and SmartPLS-3. Results are valid.
Additional comments: The manuscript was revised as recommended.


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

Input:
Title: EVALUATION OF STRUCTURAL AND ULTRASTRUCTURAL CHANGES IN THYROID AND PARATHYROID GLANDS AFTER NEAR INFRARED IRRADIATION: STUDY ON AN ANIMAL MODEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: English language should be revised in some sections of the article, mainly: Seldom the word “on” is used when “in” should be applied, some phrases are long and unclear, the word “also” is inappropriately used in some phrases.
Abstract: row 24, 26-30, 35-38, 42
Intro: 54, 66-71, 77-78, 84-88, 91
Materials and methods: 178 there is a double “buffered in”
Results: 203 the word respectively should be dropped
Discussion: 220-224
Conclusions: 240-241
Intro & background to show context.
The introduction and background of the paper are adequate. The authors present the issue with great detail and accuracy. Row 69-73 state: “In the literature, works on the effects of utilization of near infrared radiation for diagnostics and localization of thyroid and parathyroid glands are scarce, existing however some publications about the effects of this radiation when used with therapeutic intention on dental medicine”. In my opinion, this phrase is misleading. There are scars to nill literature on the effects of NIR on the structure and function of tissues. However, several papers are dealing with the use of this method of parathyroid identification in the clinical setting and its impact on clinical practice. I think that this should be highlighted. (kim et al. surgical oncology 2020, Solorzano et al. Surgery 2021, etc)
Literature well referenced & relevant. - yes
Structure conforms to PeerJ standards, discipline norm, or improved for clarity. - yes
Figures are relevant, high quality, well labelled & described. - yes
Experimental design: Original primary research within scope of the journal.
In my opinion, the papers fit well with the scope of the journal.
Research question well defined, relevant & meaningful.
The study investigates the impact of NIR on parathyroid and thyroid tissue to assess its safety. Even though this method of parathyroid identification is FDA approved and in current use, I didn’t find articles analyzing the possible impact of NIR on parathyroid tissue. It makes this paper original and relevant.
It is stated how the research fills an identified knowledge gap.
As pointed out above, the knowledge gap filed with this study should be more accurately presented, and the fact that there is scarce literature about structural changes with the use of NIR highlighted.
Rigorous investigation performed to a high technical & ethical standard. - yes
Methods described with sufficient detail & information to replicate.
 There should be a reference or an explanation after row 160-161 (times of exposure).
Validity of the findings: Meaningful replication encouraged where rationale & benefit to literature is clearly stated.
The paper cites Lerma et al. regarding thyroid structural changes. The authors found minor temporary changes on conventional histology and some minor persistent changes on electron microscopy. In the current study, no structural changes were found. Although the methods varied between the two experiments, the authors should encourage further investigation.
All underlying data have been provided; they are robust, statistically sound, & controlled.
Additional comments: I am honored to be granted the opportunity to review this article. I congratulate the authors on a fascinating and inspiring study. The topic of the investigation is original, and the literature on the matter is scarce. The paper lacks major issues. On my account, English should be revised as above. Some minor issues are present and should be addressed.


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

Input:
Title: EVALUATION OF STRUCTURAL AND ULTRASTRUCTURAL CHANGES IN THYROID AND PARATHYROID GLANDS AFTER NEAR INFRARED IRRADIATION: STUDY ON AN ANIMAL MODEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Scientific work must be formal and impersonal. Because of this, the construction of the sentence in the first or third person must be avoided.
Discussion - lines 232, 234
“We dedicated special attention to features on parathyroid mitochondria and secretory granules, without achieving any differences between irradiated and non-irradiated tissues.
We also couldn’t find nuclear or rough endoplasmic reticule alterations.”
Experimental design: It would be interesting to mention the power density of the LED that was used and the irradiated area, as the focus was approximately 20 cm from the cervical region.
A figure could be added in the text.
Validity of the findings: The discussion is very short and was not  explored. It would be interesting to include that a possible alteration of the glands may be associated with the excessive use of light for consecutive days, as we can see in the study by Parrado et al (1989). The glands were irradiated for 15 consecutive days, for example. Evaluation of changes in the densities of epithelial, colloidal and follicular volumes and the activation index revealed that the laser produces changes in the thyroid parenchyma. It was noted that there is a direct relationship between the severity of the injury and the energy of the applied irradiation.
Azevedo et al (2005) - A statistically significant hormonal level alteration between the first day and 7 days after the last irradiation was found, but no changes were found in the histological structure of the glands.
It is important to remember that the penetration of the laser is deeper than LED, even if it is the same wavelength. The LED is neither collimated nor coherent.
Additional comments: It is a very interesting study, but it could be better analyzed and explored in the discussion


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

Input:
Title: EVALUATION OF STRUCTURAL AND ULTRASTRUCTURAL CHANGES IN THYROID AND PARATHYROID GLANDS AFTER NEAR INFRARED IRRADIATION: STUDY ON AN ANIMAL MODEL
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article should be reviewed by an English speaker for proper grammar.  For instance, the authors should avoid contractions such as "couldn’t" and should use "could not" instead.

The literature references provide sufficient background and context to the studies.
Experimental design: The experimental design is good and the research question is well defined.

The numbers of rats in the study are adequate.
Validity of the findings: The finding show no structural differences in the parathyroid and thyroid glands of mice undergoing near infrared exposure for autofluorescence versus control mice.

However, the authors should include a paragraph in the Discussion section regarding the limitations of the study.  For instance, although there were no structural differences between groups, the authors did not assess thyroid or parathyroid function in the rats and therefore cannot comment on the the possibility of functional alterations of the glands from radiation exposure.
Additional comments: No additional comments


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

Input:
Title: EVALUATION OF STRUCTURAL AND ULTRASTRUCTURAL CHANGES IN THYROID AND PARATHYROID GLANDS AFTER NEAR INFRARED IRRADIATION: STUDY ON AN ANIMAL MODEL
Review round: 2
Reviewer: 1

Output:
Basic reporting: Self-contained with relevant results to hypotheses.
Experimental design: Methods described with sufficient detail and information to replicate
Validity of the findings: Conclusions are well stated, linked to original research question and limited to supporting results
Additional comments: I would like to congratulate you for reviewing the text. This is a study of great relevance.


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: EATING FOR NUMBING: A COMMUNITY-BASED STUDY OF TRAUMA EXPOSURE, EMOTION DYSREGULATION, DISSOCIATION, BODY DISSATISFACTION AND EATING DISORDER SYMPTOMS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall the manuscript is well-written, concise, and easy to read. The literature references appear sufficient, given the wide range of topics included in this study (trauma, eating disorders, dissociation, emotion regulation). The raw data is included and shared. The tables and figures are helpful to include. The hypotheses are clearly defined and described within the text.

Please consider the use of person-first language throughout the manuscript; for example, instead of "obese participants" please consider revising to "participants with obesity". Also please add labels to BMI descriptors (e.g., kg/m2). Finally, there is an incomplete sentence listed in line 374.
Experimental design: Please consider updating the manuscript to address the following:

1. How were missing data handled?
2. Some of the EDE-Q data in the SPSS file includes responses such as "7". These are not standard item-level scores for the EDE-Q measure (responses are on a 0-6 scale). Were these scores adjusted in some way? Did the authors intend for these codes to represent missing data? Please be certain these were not accidentally included in total scores for this measure. There are no 0's on this measure for any participant, which seems unlikely. It is very easy to have a 0-6 scale output 1-7 in data software. Yet, this will artificially inflate scores and pathology.
3. The DERS and EDE-Q have multiple subscales. Did the authors just use the total/global score for each of these measures? What scores was included in the analyses? Please clarify within the text to reduce confusion and enhance reproducibility.
4. How was BMI divided into three groups? Which cut-offs were used for each category? Please add additional detail about these analyses.
5. Do the authors have any further information as to the "other" category of traumatic experiences, as this was endorsed by 56.38% of the sample?
Validity of the findings: Once the data has been verified as accurate (see concern regarding the scoring of the EDE-Q measure as noted above), it may be too far of a speculation to ascertain that "eating for numbing" may explain the results of this cross-sectional study. The study did not collect any information about eating patterns or eating for numbing, outside of the EDE-Q which does not directly assess for this behavior. The title of the article implies that the researchers found a relationship with the construct of "eating for numbing" and trauma exposure, emotion dysregulation, dissociation, and body dissatisfaction. Please consider tempering conclusions or editing the title of the manuscript to more accurately represent the study's findings.
Additional comments: No additional comments


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

Input:
Title: EATING FOR NUMBING: A COMMUNITY-BASED STUDY OF TRAUMA EXPOSURE, EMOTION DYSREGULATION, DISSOCIATION, BODY DISSATISFACTION AND EATING DISORDER SYMPTOMS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. My main concern pertains to how the rationale for the current study is conveyed. Given that the current study is examining the role of factors in eating disorder symptomatology, I suggest framing the rationale for the current study from the perspective of eating disorder symptomatology (i.e., introductory paragraph should be outlining the problem of elevated eating disorder symptomatology and the following paragraphs describe the potential vulnerability factors).
2. It would be valuable to include any relevant theoretical models that provide basis for the proposed hypotheses.
3. In order to facilitate a more coherent and logical flow of ideas in the Introduction, for each paragraph, I recommend implementing clear opening and concluding sentences.
Experimental design: 1. The gaps in the literature are not clearly identified. For example, it needs to be more clearly conveyed that prior research implicates the role of trauma exposure in eating disorder symptomatology, however, it remains uncertain the mechanisms that might underlie this relationship, before then proceeding to describe these mechanisms (i.e., dissociation, emotion dysregulation). I also recommend making clear inferences that then give rise to your hypotheses (e.g., given research showing that X is related to Y and Y is related to B, this gives rise to the possibility that X indirectly predicts B through Y).
Validity of the findings: There are several sections of the results, which need to be clarified. I address each of these below:
1.	“To explore the effects of BMI on the various indices, we divided our sample into three groups according to their BMI…” – please clarify the number of people per group. Additionally, it is not clear to me which variables the groups differed on exactly and which variables they did not differ on.
2.	Regarding the first hierarchical analysis, please clarify the reasoning behind adding variables at the different steps (e.g., why was BMI added in second step, frequency of exposure to traumatic events & dissociation in the third step, etc.). Furthermore, when describing these results I suggest using the terminology “uniquely associated with” e.g., “As can be seen from Table 4, BMI, dissociation, emotional dysregulation, and BD were all uniquely associated with ED symptoms”
3.	Regarding the SEM model, please clarify the indirect associations. That is, you mention that exposure to traumatic events predicted ED symptoms through dissociation, emotional dysregulation, and BMI. However, it is unclear to me whether this was a sequentially mediated relationship or whether these reflect individual relationships (e.g., exposure trauma  dissociation  ED symptoms).
4.	Regarding the moderation analyses, as presently described, it is currently unclear to me the observed pattern of results. Given that BMI is the moderator, the pattern of relationships should be clearly described in terms of high/low BMI. For example, the relationship between trauma exposure and BD/ED symptoms was significant only for people with high/low BMI. Please clarify.
Additional comments: Here are some additional minor comments for the authors’ consideration.

-	Method, Description of EDE-Q: Please specify whether the measure of ED symptoms was the “global EDE-Q score” and how this was computed (e.g., average of the four subscales).
-	Related to one of my previous comments, in the discussion, please clarify the nature of the results which concern the moderating role of BMI, ensuring that these are clearly described in terms of high/low BMI.
-	I recommend authors expand on the implications of the rather interesting finding that trauma exposure was found to indirectly associate with ED symptoms e.g., it suggests that trauma exposure leads to greater dissociation, which in turn might lead to greater ED symptoms….
-	Additionally, I recommend authors expand on the theoretical and clinical implications of the obtained findings.
-	On a final note, another limitation worth noting is that, given the cross-sectional nature of the current study, we are unable to draw conclusions about the causal nature of the observed relationships.


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: EATING FOR NUMBING: A COMMUNITY-BASED STUDY OF TRAUMA EXPOSURE, EMOTION DYSREGULATION, DISSOCIATION, BODY DISSATISFACTION AND EATING DISORDER SYMPTOMS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Please remember to upload the correct data set in the supplementary materials. The only data set I can see through the review portal still has the incorrect scoring for the EDE-Q (1-7 instead of 0-6). It is possible that I may not be able to see the updated data set, in which case please ignore this comment.
Experimental design: Thank you for adding additional information regarding missing data, updates to the statistical analysis section, and clarification around the use of total scores rather than subscale scores for the measure. The EDE-Q global score is now clearly defined. One small note - recommend referring to the EDE-Q score throughout the manuscript and tables/figures as "global" and not "total" score. The use of "global score" is more consistent with the eating disorder literature.
Validity of the findings: The updated theoretical rationale and discussion edits have improved the manuscript. The additional limitations added into the discussion are helpful in clarifying the results from this study.
Additional comments: The authors were quite responsive to feedback. The edits have strengthened the manuscript and provide additional clarity where warranted. No further edits beyond what was mentioned above are requested at this time.


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: EATING FOR NUMBING: A COMMUNITY-BASED STUDY OF TRAUMA EXPOSURE, EMOTION DYSREGULATION, DISSOCIATION, BODY DISSATISFACTION AND EATING DISORDER SYMPTOMS
Review round: 2
Reviewer: 2

Output:
Basic reporting: Overall, the authors have done a good job in revising the manuscript and strengthening the rationale for the current study.

I have only one minor suggestion concerning the final sentence of the opening paragraph (i.e., "the current study focuses on..."). Given the importance of this sentence in setting the scene for the current study, I suggest breaking it down into two sentences to improve clarity. e.g.,

The current study seeks to test one potential account of disordered eating hypothesising that exposure to trauma, the absence of effective regulation of negative emotions, and or in the presence of dissociation, contributes to disordered eating behaviours. Such behaviours might serve to numb the unprocessed distress which arise from the trauma the individual has experienced.
Experimental design: No comment.
Validity of the findings: The nature of the results revealed by the moderation analyses are still not clear to me. These analyses seeks to tests whether BMI moderates the relationship between frequency of traumatic events and ED symptoms/BD. Thus, the result should be framed in terms of nature of the relationship between frequency of traumatic events and ED symptoms/BD at low/high levels of BMI (e.g., the relationship between x and y is and positive at high levels of the moderator). To clarify the moderation (interaction) effect, I suggest conducting follow-up analyses to estimate the conditional effects of the predictor variable on the outcome variable at low and high values of the moderator, including relevant statistics (t, p, and b value).

Furthermore, I am concerned that the authors do not fully understand the SEM results, i.e., the statement in the discussion “our model reflects what we believe may be a sequentially mediated relationship”. Please confirm whether the data supports indirect effects.
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: SIGNATURE IDENTIFICATION OF RELAPSE-RELATED OVERALL SURVIVAL OF EARLY LUNG ADENOCARCINOMA AFTER RADICAL SURGERY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall, the manuscript writing is clear and easy to understand, except some examples where the language could be improved for better comprehension, e.g. line 71-72, “We found that some patients…or patients with multiple…”; line 75-76, “on the other hand…after the first operation”.
Experimental design: I have a question on the selected TCGA RNA-seq dataset for differential expression (DE) analysis. Are the LUAD patients developed recurrent LUAD after their 1st operation? If not, why the DEGs between LUAD vs. normal lung tissue could be used for identifying prognostic signature? My understanding is, the DEGs should come from comparing the post-operative LUAD patients who showed recurrence vs. the post-operative LUAD patients who did not. Although, if the authors could provide more evidence that the LUAD vs. normal DEGs have direct correlation with prognostic recurrence, it’d be more logical to doing DE analysis between LUAD and normal lung tissues. A side note: I do notice that the authors later used a more relapse-related GSE5081 dataset for further screening. This dataset fits better to the signature identification target.
Validity of the findings: w.r.t external validation, is the high/low risk cut-off value selected in the validation set or the training set? Most common practice for biomarker identification is, these types of hyper parameters were determined in the training set for independent prediction model validation. If there is batch effect between the training set and validation set, I would recommend the authors to consider using the R package sva and removing the batch effect before model validation.
Additional comments: In the linear function of risk score, gene “PRR11” has >2x weight than the other three genes. I’m curious if the authors have dived deeper to explain why PRR11 is the driver gene among the 4 genes?


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: SIGNATURE IDENTIFICATION OF RELAPSE-RELATED OVERALL SURVIVAL OF EARLY LUNG ADENOCARCINOMA AFTER RADICAL SURGERY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Overall, the design and analysis of the study is sound, with clear results and relatively straightforward conclusions. Regarding reporting, I recommend the following to improve readability of the manuscripts:

Line 62:  the sentence "accelerated the pace of national physical examination" is bit unclear. It if is referring to a specific nation, then it should state it more explicitly. Recommend rephrasing.

LIne71- Line80 this seems like a very lengthy discussion about second operation, which isn't directly relevant to the focus of the paper. Recommend streamlining.

Line84 - Line96 Similarly, this part seems like a very lengthy introduction of the methodology. This part could be streamlined or details moved to methods section.

Line197: Description of the text does not match the figure. The text is referring Fig2E.

Line202: Description of the text does not match the figure. The text is referring to Fig2D. Fig2D and Fig2E should be switched according to the text.

Line214:  Fig3A-C y axis title can be modified to reflect overall survival, relapse-free survival and overall survival respectively.

Line218-219 Readers could benefit from having more context on what GSE50081 study has already done.

Line228: Fig3D and Fig4B should each be labelled with a title to allow reader to more easily distinguish the difference. One is for overall survival, the other one is for recurrence.

Line248: the axis ticks, labels are all very small and hard to read. Recommend enlarging text size.

Line248-249: It would be very helpful if the authors can guide readers on these figures, and provide more descriptive details on how they interpreted Figures 5D-5I before reaching conclusions.

Most of the figure legend main titles could benefit from being more descriptive. Recommend adding key information and takeaways for figure legend main titles. Especially for Fig3-Fig8.
Experimental design: Line175: In general, the software/packages versions should also be reported for reproducibility. Which R version is used?

Line176: Is the p value referred here after correction for multiple testing? If so what method (FDR, BH etc.)

Line187: the font size of the pathway text are very small and hard to read. Recommend increasing text font size.

Line208 and Line280: In the supplementary text1, the authors showed the normality test and p values, however it would improve clarity if they can also show how they determined risk score cut-off values based on normality test.

Line203-line204: Could the authors explain how the four genes were screened out from the K-M analysis. How and why were these four genes chosen for downstream signature analysis? There seems a missing methodology step here.
Validity of the findings: Overall well done study from the authors. The data analysis and results justifies the conclusions.
Additional comments: The prognostic and recurrence biomarkers has great values, and I believe this study will be of interest to scientists and clinicians involved in lung adenocarcinoma research as well as cancer biology in general. There are some areas the authors could modify to improve readability and clarity, as outlined above.


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

Input:
Title: SIGNATURE IDENTIFICATION OF RELAPSE-RELATED OVERALL SURVIVAL OF EARLY LUNG ADENOCARCINOMA AFTER RADICAL SURGERY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Much improved readibility and flow of paper.
Experimental design: The updated and optimized study design has made the study more sound and rigorous.
Validity of the findings: no comment.
Additional comments: Great work and much improved compared to the original manuscript after editing. I recommend it for publication.


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

Input:
Title: GLOBAL WARMING AND NEUROLOGICAL PRACTICE: SYSTEMATIC REVIEW
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is Clear and unambiguous, professional English used throughout.
Only minor English grammatical error noted
Line 371 - were did not ----- to delete were
Line 400 Also, to change to In addition,
Experimental design: Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.
Methods described with sufficient detail & information to replicate
Validity of the findings: Conclusions are well stated, linked to original research question & limited to supporting results.
Additional comments: Weldone. Thank you for the wonderful effort to look into the impact of temperature of climate to various neurological illness. Future directions or discussion in subsequent papers can be look into the effect of temperature to mitochondrial function. Possible explanation for the outcome could be related to mitochondrial dysfunction following climate change.


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: GLOBAL WARMING AND NEUROLOGICAL PRACTICE: SYSTEMATIC REVIEW
Review round: 1
Reviewer: 2

Output:
Basic reporting: The Introduction would benefit from review of the major points relating to the subject at hand, which can be found in Reis et al. Climate change: What neurologists should know. In Chopra J, Sawhney MS, Neurology in Tropics, 2nd ed, Elsevier, 2015, pp. 923-932. The Introduction should include a summary of the influences of temperature change (re global warming) and poor nutrition (re migrants) on the human nervous system. The discussion on MS would benefit from a brief summary of the established differential geographic distribution of this disease (see also: https://neurosciencenews.com/climate-change-brain-15806/).
Experimental design: Experimental design and data validity is adequate except that “quantitative statistical analysis” was not employed, which stands in contrast to the major conclusion that significant heterogeneity exists across studies with respect to methodology, outcome measures, confounders and study design.
Validity of the findings: The main objective of the study is to identify how ambient temperatures influence the epidemiology and clinical aspects of major [groups of] neurological disorders. The second stated objective is to investigate the prevalence and incidence of neurological disorders in human refugee and migrant populations. To the extent that the authors investigated certain disorders (Alzheimer’s and non-Alzheimer’s dementia, epilepsy, headache/migraine, multiple sclerosis, Parkinson’s disease, stroke, and tick-borne encephalitis), the review is adequate.

Acceptable, but see below.
Additional comments: There are four major reservations: (a) there is no statement of the country/continent of origin of the studies (except for tick-borne encephalitis); (b) whether the studies analyzed are generalizable because they are/are not representative of high-income, middle-income and/or low-income populations; (c) why nutritional disorders are omitted given that both climate change and migration will impact diet; for example,(i) greater dependence in sub-Saharan Africa and South Asia on environmentally tolerant plants that harbor toxins with neurotoxic potential and (ii) increased contamination of seafood from algal blooms that are increasing worldwide in association with global warming; and (d) why neuroinfectious disorders are limited to tick-borne encephalitis when data for more important disorders that involve the nervous system, such as malaria, must exist.

English usage is correct, with some exceptions: “United Nations has” (not “United Nations have”; “data are” (not “data is”) , and “Japanese migrants” (not “migrated Japanese”).


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: GLOBAL WARMING AND NEUROLOGICAL PRACTICE: SYSTEMATIC REVIEW
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have made some edits but more work is required before this manuscript meets an acceptable standard.

The major issue is the limitation in the applicability of the findings to high-income countries. There is obvious and unfortunate irony in recognizing that global health is and will continue to be impacted by global warming in commencing an analysis of the subject that essentially is limited to globally non-representative high-income populations: (https://www.pewresearch.org/global/2015/07/08/mapping-the-global-population-how-many-live-on-how-much-and-where/)  Similarly, neurological health among tiny numbers of international migrants is analyzed relative to global estimates of the number of international migrants, which amounted to 3.5% of the world’s population in 2019, with annual estimated growth rates of 3-4% [IOM World Migration Report 2020]. To a large extent, the authors will have been forced down these narrow pathways by the equally limited scope of published studies, but failure to recognize these limitations is very disappointing. At a minimum, these study limitations, plus the fact that the present findings are relevant to neurology in high-income countries, should be recognized in the Abstract, Discussion, and Conclusion.

The authors have added a reference (#6) and summarized in the content thus: ”The effects of environmental factors on neurological practice are multifaceted and complex, involving climate, temperature, biodiversity, toxins and differences in human habitation and many others.” Food availability, quality and nutritional status should be added. Note that the authors “excluded commentaries, summaries, reviews, editorials, animal studies, toxicological studies,…

Several small edits are still needed.  Prominent among these are (a) imprecise statements such as “worsen” and (b) use of “hot”, “warm” and “warmer” to define “temperature”, which should be with defined stated limits as: “high”, “higher”, “low”, “lower”, etc.

Lines
213 deficits measured by a walking test slightly worsened  [meaning?]
353 worsen cognitive symptoms  [meaning?]
215 reported worsening of their symptoms  [meaning?]
223 worsening of symptoms in MS patients during warm periods. [meaning?]
180, 365, 489 hot temperature  [see above]
313  9 mio  [?]
318 the younger [the] age
340 Although the United Nations have [has]
288, 353 warmer temperatures  [see above]
365 hot tmperatures  [see above]
360 As to [for] epilepsy
29, 275, 380 warm temperature  [see above]
418 “Most studies did not account for factors that may influence temperatures such as humidity, air pollution and geography.”  Is this correct?  Certainly, these factors influence air quality and oxygen content, but do they “influence ambient temperature”.
Reference citations employ style differences in use of initial upper/lower case in titles: e.g. 123 cf 124; 117 cf 118, and 108 cf 109
Experimental design: See above
Validity of the findings: See above
Additional comments: see above


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

Input:
Title: GLOBAL WARMING AND NEUROLOGICAL PRACTICE: SYSTEMATIC REVIEW
Review round: 3
Reviewer: 1

Output:
Basic reporting: Acceptable
Experimental design: Acceptable
Validity of the findings: Acceptable
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: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 1
Reviewer: 1

Output:
Basic reporting: -	The article is written in clear language, and the structure of the submitted article conforms to the suggested format. The introduction clearly states the objectives of the study and places the study in context. The following specific lines/tables are noted for minor corrections, which would improve the paper’s clarity.

75-76: There is a statement here that should be referenced.

123: If known, the concentration of Lasalocid in the final TMR should be given, rather than the concentration in the pre-mix.

131: The use of D9-Clenbuterol in this study should be explained (is this a laboratory control?)

198-200: The number of cuts made is not easy to understand based on these sentences. Were there five cuts made on the darted side and one on the non-darted side?

249, 252 and Table 5: These both mention the testing of striploins. Were striploins tested or bottom rounds? There is no mention of collecting striploins in the experimental design.

273-274: A mean value is given (0.13mcg/mL) that is not within the range of individual values given (0.160-0.238mcg/mL). Using the table it appears that the min should be 0.016 and max 0.241.

Table 4: For the values L*, a*, and b*, it appears that the same asterisk used for KPH applies to these values, a different symbol should be used for KPH.

Table 7: Is the “dart” category just one value (that of the darted steak) or an average of all 5 steaks on the darted side?

Figures 1-3 and Tables 1-3: These all state in the title “See text for explanation of dosing”, yet an explanation of dosing was not easily found in the text. A specific page, line, or section reference should be made.

No specific location:
-	The format for reporting p values should be consistent throughout
o	Italicized vs normal text
o	Spaces between the p, the sign, and the number vs no spaces
o	Same number of significant digits throughout
-	Would it be possible to combine tables 6 and 7, maybe even table 5?
-	In the tables the treatments are inconsistently labeled (some spelled out, others abbreviated)
Experimental design: -	The research question is clearly defined, and is relevant and meaningful. The knowledge gap being investigated is identified.

-	The methods are consistent with ethical standards in bovine veterinary research; however, the illegal use of CCFA should be more clearly addressed as to why it was included in the study, reiterate the fact it’s illegal, describe whether these animals entered the food supply, etc.

-	There are some areas of the methods section where more detail is required, both to provide confidence in the technical standard of the research, to promote reproducibility, and allow readers to appropriately interpret the results.

o	Overall, there appears to be sufficient detail for reproducibility in the laboratory methods (both pharmacokinetic and post-mortem carcass quality and tenderness assays), however, more detail in the methods section regarding the live animal phase of the experiment would be useful. It would be difficult to reproduce the live animal phase from the current text.

o	The description of the blocking of animals and assignment to treatment groups is well done, however it is unclear to the reader how the final numbers in the pharmacokinetic study were reached. It is evident that more animals were randomly assigned to treatment groups than were analysed - forty steers were randomly assigned, yet only 5-6 per drug treatment, for a total of 17, had blood samples measured. It is unclear how this subset of animals was selected for analysis; were they randomized? Additionally, 3 animals were injected with ceftiofur at a later time, but the reason is not explained in the manuscript. More details should be provided in the methods and results section outlining the criteria used to select animals for analysis, and details should be provided about the origin of the additional 3 animals in the ceftiofur group, as well as the rationale for adding them to the study.

o	For drug delivery, some details are provided on the range, needle type, and intended target, but more detail would be required to reproduce the experiment. A good example of a methods section that provides this level of detail can be found in Coatzee et. al. (referenced in the study). For examples of some questions while reading the manuscript: What was the gun’s model and what pressure was it charged to? Were darts recovered and checked for discharge of their contents? Did any darts hit the animal in an unintended location, and if so, was this animal still used for either pharmacokinetic or meat tenderness analysis? Did any needles remain lodged in the tissue of an animal, and if so, for how long, and what was the procedure for removing them? If available, providing these details would aid reproducibility and improve clarity. If not available, the authors should state that they are unknown or not recorded.

o	Drugs and drug doses, while listed in the respective tables, should also be listed in the methods section. Rather than saying “Each drug was administered to 10 steers…” on line 114, the authors need to actually describe the experimental groups as that is fundamental to understanding the study.

o	With regards to identifying the injection site for analysis of the meat (line 197), no mention is made of the observer being blinded to the treatment. If the observer was blind to the treatment group, this should be stated to improve confidence that no bias was inadvertently introduced in selection of the meat samples. If the laboratory personnel processing the meat and performing the shear force testing were blind to the treatment group, this should also be stated.
Validity of the findings: -	The conclusion of the study is: “Delivery of ceftiofur crystalline free acid, tildipirosin, and tulathromycin to cattle using pressure-adjustable pneumatic gas-powered dart gun may result in altered pharmacokinetics and therefore altered meat withdrawal times and altered therapeutic efficacy. Delivery of tildipirosin and tulathromycin to cattle with dart gun may also result in detectable decreases in tenderness of harvested steaks” (lines 363-367). This conclusion is reasonably supported by the data, however, it is recommended that the following points are addressed in both the discussion and conclusion sections:

o	The experiments are not controlled in a way that allows the pneumatic dart delivery method to be compared with conventional injection methods for the same drugs, since there are no conventionally injected animals in this study to serve as positive controls. The authors do acknowledge that there are several reasons, including dose and sampling time, that could account for the differences between their research and existing published studies (lines 297-299), but more information should be provided in the discussion section about the reason(s) for not including these controls in the study, the limitations of comparing data obtained in this study with that of previous studies, and opportunities for future research.

o	Recommend more information in the discussion section comparing the methods used in this study with those of the previous pharmacokinetic studies that are referenced in this manuscript.

o	Since the study design does not allow for direct comparison between conventional and dart delivery injection methods, the conclusions should avoid comparative terms such as “altered”, or should state that meat tenderness/pharmacokinetic data may be altered when compared with existing data from previous studies.

-	Since the injection site recommended by the BQA is in the neck, more information in the discussion section about the rationale for performing injections in the bottom round would be very informative. For example, was this site selected because it is this a common site of injection with pneumatic darts among producers, in the authors’ experience?

-	A brief discussion of how the results from the pilot study informed the design of the main study would also aid the reader’s understanding of this paper, particularly with respect to the use of flunixin. Without discussion, it is not clear to the reader why this drug was used. A short section on the pilot study should be included in the discussion.

-	Related to the above point, the second objective of the study (evaluation of injection site reactions at 13 days) seems to have been completed in the pilot study only. Therefore, a discussion of how these results informed the design of the main study would be very informative.
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: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is overall well written and worthy of publication (after corrections and improvements). Adequate background information is given to demonstrate that the information conveyed in this manuscript is valuable to the beef industry and veterinary profession. There are several areas where clarity could be improved, some portions where additional information is needed, and at least one notable typographical error.

Specific suggestions or points of concern:
Line 72: Would suggest clarifying “The ANTIMICROBIAL products used in this study….”
Line 75: “….previous studies have shown that…..” Provide citation for these studies.
Line 79: “… urge that all injectable products should be administered subcutaneously rather than intramuscularly” (I suggest adding “if permitted by drug label”).
Line 115: Device is described as “pressure adjustable.” Please describe how the pressure to be used was determined, and whether it was consistent across the study.
Line 138: Reference made to SPE prior to the acronym being defined (line 141).
Line 229: It is unclear why results for drug concentrations are not reported in results, but rather primarily included in the discussion.
Line 273-274: Statement is made that mean Cmax is 0.13, with a range of 0.160 to 0.238. The mean isn’t within the reported range. Looking at table 1, it appears the range should be 0.016 to 0.238.
Line 277: Statement of …”24 hrs but samples only collected at 2 and 24 hr….” is confusing.
Line 297: It is stated the other study sampled more intensely early in the dosing period. Did Cmax occur during those early sampling intervals?
Line 330: “…but sampling times were different (in the present study?)….”
Various: Interpretation of “similarity/dissimilarity to other publications” and “widely variable” seems subjective and perhaps inconsistent. For example, line 274: “…wide range of individual values.” This reviewer doesn’t view that range as particularly wide. The assessment that median Tmax was similar to other reports is open to interpretation- 8 hours seems notably different than 18 hours (Washburn) or 24 hours (Witte- although it is noted that sampling was only done at 2 and 24 hours). This is apparent again in lines 314-317, where tulathromycin is in the range of previous reports, where previous reports range from 277 to 1820. Such a range seems quite broad, and thus it is expected that the current study would fall within this range. If 0.160 – 0.238 is considered a wide range of individual values (line 274), then 277 to 1820 should likely be identified as extremely broad, and effort made to explain why such a range should exist, and the implications of such for the current study [as noted above, the range of 0.16 to 0.238 is apparently in error].

Table 1: Various terms in the legend don’t correspond to the terms in the table. For example, table includes the term “AUClast”, which would seem to correspond to the description in the legend for “AUC0-obs.” Similarly, there is no description of the term “AUCINF_obs”, which is used in the table. This occurs for nearly all of the terms.
Discussion information for ceftiofur is reported as mcg/ml in the text, but as ng/ml in the table.
Experimental design: It is unclear what inclusion of the pilot study contributes to the manuscript. Of particular concern is the appropriateness and relevance of administering 10 ml flunixin meglumine. This is a very large volume (exceeding even the high end of the label dose) of a well-known irritating compound. Also questionable is the use of 5 ml of CCFA at a dose at or below label for these animals. Additional information and context should be provided if the pilot study is to remain part of the manuscript.
Validity of the findings: My greatest concern regarding the entire paper is in the statistical methods. I consider the failure to adjust for multiple comparisons inappropriate and misleading. Specifically, under “shear force data analysis,” it is stated that a t-test between means was done (presumably only for those where a p<0.05 was found for the group, although this should be stated). Using a t-test for multiple comparisons greatly increases the likelihood of a type I error. As an example, the p-value for dressing percentage is 0.05. It is difficult to explain why the treatments would notably impact dressing percentage, and this p-value is “marginal.” It would be expected in such cases that post-hoc pair-wise comparisons would not find a significant difference between most (or possibly all) pairs. That isn’t the case when an unadjusted t-test is used, resulting in (likely capricious) “significant differences” between the various groups. It is unclear, but possible that an even more notable statistical error was made regarding table 7. If the overall p-value is 0.08, then no post-hoc between treatment comparisons should be done. Post-hoc pairwise comparisons should only be done when initial analysis finds a statistically significant difference between treatments, and under that circumstance, pairwise comparisons must account for the increased possibility of an error due to multiple comparisons.

It is unclear what the definition of “outlier” is, and thus exclusion of individual animals from various analyses seems arbitrary. This should be clarified, and ensure that consistent standards are applied in all cases. Particularly given the complexity of pharmacokinetic data, it is difficult for a reader to determine whether outliers truly represent an anomaly they are unlikely to deal with, or whether the outliers occurred due to unknown variability with the delivery method. If it is the latter, then the practitioner should know that darting can produce such variability, and that values obtained in practice will share this variability.

Reporting of results in the table also gives me concern. I am not familiar with previous studies examining ceftiofur metabolism, and thus I am uncertain of number of subjects and range of variability commonly seen. However, given the extreme variability and the relatively small sample size in the present study, I question the appropriateness of presenting mean values for Cmax, and comparison of those values to ones reported previously. There is a similar concern would exist for AUMC parameters (where the coefficient of variation is >50%). Indeed, a median value is reported for Tmax, but no explanation is provided as to justification/rationale for reporting median here but not the other parameters. While I concede that inclusion of individual animal data helps the reader to interpret some of this, I believe it would likely be worthwhile to include median values. Discussion should also be offered as to the appropriateness of relying upon mean values as opposed to medians (particularly vis a vis the expected variation, given results from other studies).
Additional comments: I am in favor of acceptance of the manuscript, if authors address concerns noted above. Of particular concern are issues related to statistical methods, and reporting of results.


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

Input:
Title: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript is considerably improved from the first version; however, there are still 5 outstanding items to address )Items 1, 2, 3, 5, and 6), as well as one new comment (Item 4). These are described here and/or in the following sections as appropriate, with the Original Reviewer Comment, the Author Reply, and then New Comments from the Reviewer.

1. Original Reviewer Comment - Since the study design does not allow for direct comparison between conventional and dart delivery injection methods, the conclusions should avoid comparative terms such as “altered”, or should state that meat tenderness/pharmacokinetic data may be altered when compared with existing data from previous studies.

Author Reply - This is a fair point, so we’ve revised the conclusion.

New Comments from the Reviewer - The abstract should contain a similar edit, as it is unchanged from the original version. In the reviewer’s opinion, a more appropriate conclusion would be to simply state that this study describes the pharmacokinetic properties of four drugs administered by a pressure-adjustable pneumatic gas-powered dart gun, thus avoiding comparative statements entirely.

2. Original Reviewer Comment - The format for reporting p values should be consistent throughout
o Italicized vs normal text
o Spaces between the p, the sign, and the number vs no spaces
o Same number of significant digits throughout

Author Reply - Changes made as suggested.

New Comments from the Reviewers - Some changes have been made but there are still some inconsistencies. P-values are sometimes given as an exact number (p=0.003, line 271) and sometimes given as a “less than” or “greater than” (p>0.05, line 268). Either format is appropriate, but it should be consistent. It is generally accepted to report exact p-values for all results except those below 0.001, in which case p < 0.001 is appropriate.
Sometimes the p-value is capitalized (line 258) and sometimes it is lowercase (line 268). This should be consistent. Sometimes the symbol p is directly adjacent to the greater-than sign (line 268) and sometimes there is a space between them (line 274). The format should be consistent.

3. Original Reviewer Comment - Would it be possible to combine tables 6 and 7, maybe even table 5?

Author Reply - I combined the tables and hopefully these look better.

New Comments from the Reviewers - Most of the data from Tables 6 and 7 have been added to Table 5 – it looks much better. Table 7 is now redundant and should be removed from the manuscript. Table 6 has some data on position of the bottom-round steaks that can also be moved to Table 5, eliminating the need for table 6 as well.

The column names “Control” and “Darted” in Table 5 appear to be associated with wrong W-B SF values below them (i.e., they have been switched during reformat of the tables).

The explanation of the positions of the bottom-round steaks on the darted and non-darted side in the methods section has improved greatly from the original version, as has the position of the top loin steaks from the anterior edge (presumably referred to as positions 1, 2, 3, and 4 in Table 5). It would add clarity to eth manuscript to include reference to positions 1, 2, 3, and 4 in the materials and methods, as well as footnotes to Table 5 to clearly define/describe the positions of both the bottom-round and top loin steaks, including that “Dart Side” actually refers to the presumed dart entry site on the dart side.

4. New Comment form the Reviewer - Some drugs/equipment sources are not cited completely (the company name is given, but not the city and state where the company is located).
Experimental design: 5. Original Reviewer Comment - The methods are consistent with ethical standards in bovine veterinary research; however, the illegal use of CCFA should be more clearly addressed as to why it was included in the study, reiterate the fact it’s illegal, describe whether these animals entered the food supply, etc.

Author Reply - Language about legality has been added to the methods and discussion sections.

New Comments from the Reviewers - The revised manuscript adequately emphasizes that CCFA use is illegal when administered by an unapproved route in the United States, but we still have the following concerns:

-	The authors have stated that the animals given CCFA did not enter the food supply in their rebuttal letter, but we were unable to find any point in the revised manuscript where this was stated. This should be clearly stated in the Abstract, Materials and Methods, Discussion, and Conclusions.
-	It is still unclear to the reader why CCFA was included in the study. Anecdotal reports that CCFA is sometimes delivered by dart gun illegally in the US are, in our opinion, not good reasons to study the pharmacokinetics and effects on meat quality of the drug when administered by this route. Data obtained from this study on CCFA cannot inform decision-making by producers or veterinarians in the US because the use of the drug by that method would be illegal anyway. The authors should discuss potential applications of their research into this route of administration for CCFA.
-	It is important to state precisely what is prohibited by the regulatory restrictions on the use of CCFA. The statement on lines 107-108 that “cephalosporins may not be given in an extra-label manner in the US” is not quite correct. According to the article cited, the FDA order allows for the extra-label use of cephalosporins to treat or control a disease condition that is not present on the label, for example. The relevant prohibition is specific to the use of cephalosporins by an unapproved dose level, frequency, duration, or route. The statements on illegal use of cephalosporins in the manuscript should be revised to make sure they are consistent with this reference.
-	The cited article on line 108 should be cited as (Federal Register, 2012) or (US Food and Drug Administration, 2012). Citing it as Anonymous makes the source appear less authoritative.

6. Original Reviewer Comment - The description of the blocking of animals and assignment to treatment groups is well done, however it is unclear to the reader how the final numbers in the pharmacokinetic study were reached. It is evident that more animals were randomly assigned to treatment groups than were analysed - forty steers were randomly assigned, yet only 5-6 per drug treatment, for a total of 17, had blood samples measured. It is unclear how this subset of animals was selected for analysis; were they randomized? Additionally, 3 animals were injected with ceftiofur at a later time, but the reason is not explained in the manuscript. More details should be provided in the methods and results section outlining the criteria used to select animals for analysis, and details should be provided about the origin of the additional 3 animals in the ceftiofur group, as well as the rationale for adding them to the study.

Author Reply - Methods section has been revised to clarify.

New Comments from the Reviewers - The authors should clearly explain the reasons for the apparent removal of data points between revisions. The first draft reads as though 10 animals were blocked and randomly assigned to the CCFA group at the beginning of the study. Five of these animals were included in the pharmacokinetic analysis, and three additional animals were administered ceftiofur at a later time (lines 119-120 of original version) and were also included in the pharmacokinetic analysis. A total of 13 animals appear to have been injected with CCFA in the original version of the manuscript, but only 10 are discussed in the revised version. The apparent exclusion of animals in this section between revisions requires explanation. If additional animals were not added to the study at a later time and the original manuscript was in error, the reason for the error should be clearly explained in the rebuttal letter. If additional animals were added to the study, then the details of the origin of these animals and the rationale for including them in the study should be explained in the revised manuscript.
Validity of the findings: Please see above comments in sections 1 and 2
Additional comments: Please see above comments in sections 1 and 2


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

Input:
Title: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 2
Reviewer: 2

Output:
Basic reporting: Minor recommendations for changes- see below.
Experimental design: No comment.
Validity of the findings: Line 392: It bears acknowledgement that pharmacokinetics, withdrawal times, and clinical efficacy may differ for all three products when delivered via dart as compared to conventional delivery (as you’ve done). Nonetheless, I find it notable that tildipirosin had much less extreme variability in several of the pharmacokinetic parameters, compared to that seen with CCFA and tulathromycin.  This is most clear for Cmax and Tmax. I suggest you consider addressing this in the discussion or conclusions. If I had to make a decision on drug selection based upon this study, I would choose tildipirosin. Is that decision defensible?
Additional comments: Line 94: Provide manufacturer info. for Zuprevo
Line 99: How was the determination of need for 104 psi made? Is that based upon dart manufacturers’ recommendations, as determined by volume? Or from a previous report? Or based upon trial and error process?
Line 116: standardize phrasing and punctuation for the groups (there’s a dash after “ceftiofur, but not after tulathromycin).
Lines 254-255: “…dressing percentage and L* values were significant sources of variation within treatment.” This would generally mean that, when inserted as a covariable, dressing percentage and L* values altered the impact of treatment on a third variable. From examining table 4, I do not believe that is what is meant by this sentence. My interpretation of table 4 is that “…dressing percentage and L* values were significantly different between treatment groups.” Or “…dressing percentage and L* values differed significantly across treatment groups.” Or “….significant differences in dressing percentage and L* values were found among treatments.”
Line 256: I’m surprised that an ANOVA value for the full group comparison can be exactly equal to the value found after post-hoc correction for multiple comparisons. It is also surprising that the p-value is exactly the same for two post-hoc comparisons (SALN vs. CCFA and SALN vs. TILD, particularly considering the mean dressing % varied by 0.5% between CCCFA and TILD. Please confirm these values prior to publication.
Line 296: Eliminate double period punctuation.
Line 392: It bears acknowledgement that pharmacokinetics, withdrawal times, and clinical efficacy may differ for all three products when delivered via dart as compared to conventional delivery (as you’ve done). Nonetheless, I find it notable that tildipirosin had much less extreme variability in several of the pharmacokinetic parameters, compared to that seen with CCFA and tulathromycin.  This is most clear for Cmax and Tmax. I suggest you consider addressing this in the discussion or conclusions. If I had to make a decision on drug selection based upon this study, I would choose tildipirosin. Is that decision defensible?


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: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 3
Reviewer: 1

Output:
Basic reporting: No further comments (please see "general comments" below).
Experimental design: No further comments.
Validity of the findings: No further comments.
Additional comments: Thank you very much for your changes and explanations in the letter. The manuscript is markedly improved from its earlier versions and is suitable for publication with some recommendations, outlined below. We appreciate the improvements to the methods section and clarification of the experimental numbers. These are much clearer in the manuscript’s current version.

We understand the authors’ concern about excessive repetition of the point about the illegality of extralabel use of CCFA, however it is important to include a short statement in the abstract, since some readers will only read this section. A sentence in the methods section of the abstract, perhaps on line 32 (tracked changes document), that states: “Due to the prohibition of extralabel injection routes for CCFA in the United States, the animals injected with this drug via dart gun did not enter the food supply.”, or a similar sentence to that effect, should be used.

We also understand the rationale for using P>0.05 in some cases rather than the exact P-values. For consistency, they should be written as P>0.050 (with the same number of significant figures as the exact P-values reported elsewhere in the text).

Some brief copy-editing items are as follows. The line citations refer to the tracked changes document:
o	Line 222 “top round”: We think this should read “bottom round”.
o	9 animals were injected with ceftiofur according to figure 1, but only 8 were injected in this group in the text and in table 1.
o	6.6mg/kg dose of ceftiofur stated in figure 1/table1, but only 6mg/kg was stated in the text.
o	Line 39 on table 5 should read “text” not “test”.
o	Table 6 and 7 are still cited in the discussion section after their removal.
o	The P-value on line 279 is differently formatted than the rest of the P-values in the manuscript.
o	The citation of Anonymous (the first reference) has changed in the reference section as per our recommendation but the in-text citations throughout the manuscript have not changed.

Thank you very much for your attention to these edits.


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: EFFECTS OF DELIVERY VIA PRESSURE-ADJUSTABLE PNEUMATIC GAS-POWERED DART GUN OF THREE ANTIMICROBIAL DRUGS (CEFTIOFUR CRYSTALLINE FREE ACID, TILDOPIROSIN, AND TULATHROMYCIN) ON DRUG DISPOSITION AND MEAT QUALITY IN CATTLE
Review round: 3
Reviewer: 2

Output:
Basic reporting: Line 76: “…important contributor to the consumer….” seems like an odd (and inaccurate) phrasing. Suggest: “…. Important contributor to consumer satisfaction…” or “important characteristic for the consumer….”
Line 117: “….were administered ceftiofur…” Specify the formulation was CCFA. Also specify route- was this administration per label, or administered in the round (to better compare to that administered via dart)?
Line 271: Move “(P=0.73)” from after “affected” to between “…loin” and “affected….”
Line 307: Should this be “similar” rather than “equivalent?”
Line 379: “…the ones that did…” could be modified to state “….the ones that penetrated the skin….”
Line 275 states “all steaks (from darted and non-darted sides) from cattle treated with ceftiofur and saline were more tender than from cattle treated with tulathromycin or tildipirosin.” Could this not possibly indicate a difference in the cattle themselves more so than being attributable to treatment (considering it was also seen in the non-darted sides)? This possibility isn’t mentioned (although admittedly, the discussion states that the difference may not be clinically significant).
Table 1: Tmax mean has a value of 8*, with the asterisk indicating median rather than mean. It would seem more appropriate to me to not include any value there (simply an asterisk saying “median is preferred” or similar) rather than putting an inaccurate value with an asterisk that it isn’t what it states it is.
Table 1: The legend/key refers to t1/2, but the table has t½λz. These should be the same.
Table 5: I would suggest changing layout for the p-values to be positioned elsewhere, but will defer to the editor.
Experimental design: The number of cattle enrolled in the ceftiofur treatment is inconsistent. The abstract states 8 (line 30); materials and methods states 5 (line 115) immediately followed by “Three additional steers….” (so total of 8? Although it isn’t clear that these three were handled the same way). Figure 1 legend states “Mean serum concentrations of ceftiofur…. In 9 animals”  and table 1 shows results from 8 individuals.
Validity of the findings: No comment
Additional comments: No additional comments