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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 1
Reviewer: 1

Output:
Basic reporting: There is growing body of literature regarding the role of gut microbiome in the development and progression of diabetes mellitus.
The peculiarities of bacteria presence in subjects with overt diabetes mellitus and prediabetes had been shown.
Experimental design: In the manuscript gut microbiome was examined in the fecal samples in three cohorts of subjects with overt diabetes mellitus, prediabetes and control group, 60 subjects in each group.
Validity of the findings: The aim of the study and the presented data are of interest.
Additional comments: There are some questions, which I believe have to be addressed in the preparation of the final version of the manuscript.
In the Characteristics of patients studied, there are many data, which seems to be not necessary such as levels of education or income. However, there is no information about HbA1c levels and antihyperglycemic treatment if any was prescribed. This point is important as the first class of such medications, metformin, could affect gut microbiome.
The article can be shortened especially Introduction, which would be better taken by the reader.


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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 1
Reviewer: 2

Output:
Basic reporting: English is mostly clear. However, there are some typos left.
Structure of the article corresponds to guidelines, wide range of references is used. However, I would suggest to reevaluate the Introduction section - it appears to be almost too long, some of the information has been duplicated between paragraphs (for example, lines 65 - 71). References from previous studies on microbiome of the three cohorts are missing (line 93).
Experimental design: The authors have involved and characterized a decently sized cohort of three subgroups. However, several questions need to addressed and improved regarding the data analysis.
1. Was a correction for multiple testing performed? Could you please specify the method for this and depict the adjusted values in the manuscript? Was any cofactors included in the statistical comparisons?
2. Could you please specify the tools/softwares used for beta diversity analysis?
3. What was the chosen sequencing depth?
4. Was there any data preprocessing steps performed? What tools and databases were used for that? I see some filtering info in supplementary data but the thresholds and some steps are unclear.
5. How did you ensure the immediate freezing of the samples? Were they collected in hospital environment?
6. Could you specify what method was used to disrupt the microbial cells - enzymatic or mechanistic?
Validity of the findings: The presented results are interesting and somewhat compatible with data previously reported from other studies. However, the selected cohorts unfortunately are poorly characterized and various flaws have not been discussed. For example, prediabetes ir highly various groups of patients, therefore putting all preDM patients in one subgroup could prevent from detecting true differences in the gut microbiome profiles.
In addition, despite the fact that authors acknowledge the flaw of not having any information on medications or BMI, they do not fully discuss the possibility of repeating previously characterized misinterpretation of differences between T2DM, healthy and even preDM with effects caused by antidiabetic and other(PPI, statins) medications. Could it be possible to obtain the information on medications by using some health data registries?
*it is important to note, that not having these data and not using them as cofactors for statistical analysis can lead to significant misinterpretations in the results of microbiome data comparisons!

Although various statistical comparisons have been made, the presentation of them is unclear - more precisely, it is complicated to understand at what times comparisons are performed on all subgroups together and where are specific comparisons between just two subgroups. In addition, in results section, a lot of the text is duplicating the information presented in the tables.

There is some controversy in author comments on diet data, where they characterized the diets as similar and further describe the numerous differences. I would suggest that more detailed diet data analysis is needed, most likely by inviting some nutritionist.

In discussion, it seems that results representing some controversial data to those presented by authors are missing. For example, P.copri effects on T2DM have been shown to diet and strain specific, as described by Filippis et al 2019 (https://doi.org/10.1016/j.chom.2019.01.004).
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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 1
Reviewer: 3

Output:
Basic reporting: No comments on basic reporting
Experimental design: No comments on experimental design
Validity of the findings: A Table comparing the authors’ results with the previously published data will be very helpful and would allow to evaluate the interesting findings presented in this paper and their potential significance for the onset and escalation of the disease.
Additional comments: The present study revealed significant alterations in the gut microbiome between prediabetic (PreDM), diabetic (T2DM) and non-diabetic individuals. These interesting data suggested that early intervention in PreDM patients may have implications for gut microorganisms in transition to T2DM.
The authors cited numerous publications reporting on diabetes-related alterations in microbiomes that included some groups of microorganisms which were also analyzed in this paper and produced sometimes conflicting results (e.g., lines 212-216). While discussing significant changes found in the T2DM and pre-DM microbiomes, a Table comparing the authors’ results with the previously published data will be very helpful and would allow to better validate the interesting findings presented in this paper and their potential significance for the onset and escalation of the disease.


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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 2
Reviewer: 1

Output:
Basic reporting: The same as the in the original review.
Experimental design: The same as the in the original review.
Validity of the findings: The same as the in the original review.
Additional comments: In the revised version of the manuscript authors have addressed my previous 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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: 1. The information in the manuscript and the answer to my question presents controversial information regarding the method used for microbial DNA extraction: in manuscript is says - "Total DNA of microorganisms was extracted from all 180 samples using a QIAamp stool DNA minikit (Qiagen, Germany)" (lines 125-126), however in the rebuttal letter authors have stated "DNA was extracted using the FastDNA Spin Kit for Soil from mp biomedicals.". Could the authors please clarify!

2. I would suggest to provide the sequence preprocessing descriptions with all relevant thresholds used in a supplementary file, if there is a problem with providing this info in the main manuscript (due to word limitations) - such info should not be excluded for the sake of reproducibility!

3. The authors have added the following sentence to the abstract - "..and the intestinal bacterial community was quantitatively detected by real-time fluorescence quantitative PCR." - however, there is no detailed information on such approach in the methods section. Could the authors please clarify the needed info in the methods section as well.
Validity of the findings: no comment
Additional comments: I thank the authors for extensive changes and the clarifying answers provided.


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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 2
Reviewer: 3

Output:
Basic reporting: No comments.
Experimental design: No comments.
Validity of the findings: For most of the microbial groups examined here, the data presented in the manuscript and the results from other publications appear somewhat controversial, which should be addressed and discussed.
Additional comments: The previous comments were not completely responded to, although the Table_S1 was generated by the authors. Therefore, here are some notes for the authors:

1. Table_s1 was not referred to in Results nor it was discussed (e.g., in the paragraph l.244-251).

2. The title for Table_s1 is not appropriate and should be changed, e.g. for “Diabetes-associated differences in gut microbiome abundance: comparison with previous studies” (or “Gut microbiome abundance: comparison with previous studies”).

3. Please, make the Table more accurate and stick to a certain way of marking differences, i.e. either use arrows or “lower/higher”. Currently, the authors’ results and those by Palacios et al for Bacteriodetes, are missing from Table_s1, as well as the results by Ahmad et al., 2019 for Prevotella were not included in Table_S1.
Overall, for most of the microbial groups (except for Prevotella), the data presented here and  the results from various labs appear somewhat controversial, which should be addressed and discussed.

4. The authors have added in the Introduction two new publications that reported on the same three groups of people: those with newly diagnosed T2DM, PreDM and non-diabetes, as studied in this paper (AH et al. 2020; Zhong et al. 2019; line 103) which, however, were not discussed at all in this manuscript. It would be rather appropriate if the authors could include the data from these two publications into Table_S1 and discuss them.

Correct the way the papers are cited on line 244 [ Larsen et al.(Palacios et al. 2017) found...]


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: CHARACTERISTICS OF THE GUT MICROBIOME IN PATIENTS WITH PREDIABETES AND TYPE 2 DIABETES
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Thank you for answering the comments and improving the manuscript accordingly.


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: ASSOCIATION BETWEEN URIC ACID LOWERING AND RENAL FUNCTION PROGRESSION: A LONGITUDINAL STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Proposed paper is really interesting since it explore something that is today really under discussion regarding the role of uric acid lowering. However some revision are needed before it can be accepted for pubblication. Please see below for detailed comment.
Experimental design: Some problems regarding population description (only diabetic subjects? diabetic and IFG/IGT? really population cohort?) and models are present. Please see below for detailed comment.
Validity of the findings: Some conclusions are not related to study results. Please see below for detailed comment.
Additional comments: Major comment:
- From the methods section (as well as abstract) it is not clear wether or not patients are all diabetic. This need to be specified because the name of the study define the diabetes as a charactheristic of the population however this is not specified in inclusion criteria. If yes this need to be inserted in the title and discussed in the relative section. In fact this is not a population cohort but a cohort of diabetic patients in which renal end-point could be determined by this condition itself. Furthermore, in methods section diabetes and prediabetes were defined, the prevalence of the two condition need to be presented. If prevalence of prediabetes is high the population should not be defined as a diabetic one. Confusion is now present regarding what patients are we evaluating.
- Is diabetes disease duration available? if yes this need to be described and inserted into the model.
- In the multivariate model SBP should not be inserted toghether with PP variation in order to avoid collinearity. Please insert only SBP variation. On the same way, HbA1c should be replaced with HbA1c variation.
- Why renal declined was not used as a continue variables? although categorization permit to calculate HR it determines the lost of a lot of information. On the same way also categorization of UA changes in quartiles determines a most simple way to describe findings but result in information lost. Also multivariate models with UA changes and GFR changes both as continue variables need to be presented in order to confirm the association founded with categorical variables.
- One important and recently published papers that focused on uric acid and target organ damage (with also a focus on renal damage) has been forgotten and need to be cited (J Clin Hypertens (Greenwich). 2018 Jan;20(1):193-200.)
- Although the exclusion of subjects on uric lowering therapies is described in the discussion this is not clearly stated in the methods and exclusion criteria. Furthermore, was therapy assessed at both visit or only at the first one. If it was not assessed at the second one it means that some subjects could start hypouricemic agents the day after the first visit and no one will never known. If this is the case this need to be inserted into discussion as an important limitation.
- The following sentence is completely disregarding respect to the study results: "Our study found that uric acid lowering was beneficially associated with changes in renal function, which indicated that uric acid-lowering therapy should be timely and appropriate and should be a part of the comprehensive management of chronic renal disease.". In fact if patients on this study are not treated what we found is that spontaneous variation of UA are related to variation in GFR. No conclusion regarding therapies can be done!!!

Minor comment:

- In abstract and aims section please delete the following sentence: "and to provide support for clinical strategy and community health management". In fact this is the aim of every published research and it doesn't need to be stated.
- In the abstract please clarify if quartile 1 is the one with the higher or the lower changes (conversely it will define also quartile 4) because as it stand is now unclear. The same need to be inserted also in the methods section and recal at the beginning of results section.
- Prediabetes is an incorrect definition, please use Impaired Fasting Glucose or Impaired Glucose Tollerance as appropriate.


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: ASSOCIATION BETWEEN URIC ACID LOWERING AND RENAL FUNCTION PROGRESSION: A LONGITUDINAL STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript is generally well-written, although the titles of tables 2 & 3 need to be modified (please see comments for the author).
Experimental design: This manuscript is within Aims and Scope of the journal. Research questions were well defined and meaningful. The study was performed according to ethical standard. Methods description was clear.
Validity of the findings: The association between uric acid and renal function was well documented. This study provided further information in a reasonable sized longitudinal study.
Additional comments: The authors performed a longitudinal study for uric acid change and renal function progression in the REACTION cohort. Their results showed that the decrease (or increase) of plasma uric acid levels were associated with the improved (or decreased) eGFR.

Several issues needed to be addressed before the manuscript could be accepted for publication: 1) The authors emphasized that the "lowering" uric acid was associated with changes in renal function. Actually, it was not an intervention prevention study.  The authors may need to explain the decreased uric acid level in their population, since they did not provided how many subjects had received uric acid lowering agent during the 4-year follow up. 2) The decline and improvement of renal function were simply defined by a dichotomous eGFR change. However, subtle changes in BUN and creatinine measurements could be common in any given follow-ups, less significant eGFR changes should not be interpreted as a declined or improved renal function.

A minor issue: in titles of tables 2 & 3 were a little misleading, "prevalence" are actually hazard ratios.


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: ASSOCIATION BETWEEN URIC ACID LOWERING AND RENAL FUNCTION PROGRESSION: A LONGITUDINAL STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: authors replies to all the query and paper improves.
Experimental design: experimental design is now valid
Validity of the findings: findings are now valid
Additional comments: Authors replies to all the query raised and paper signifcantly improves. It can now be accepted in its present form.


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: CHARACTERIZATION OF SPRAY-DRIED GAC ARIL EXTRACT AND ESTIMATED SHELF LIFE OF Β-CAROTENE AND LYCOPENE
Review round: 1
Reviewer: 1

Output:
Basic reporting: well described
Experimental design: well described
Validity of the findings: well described
Additional comments: the manuscript is well written and obtained good results. Congratulation!

Introduction: well described

Materials and method:
- Line 90: Rpm to g, also abstract
- Line 96-97: there are 3 constant variables, drying air flow rate, compressor air pressure and feed rate. But only 2 numbers are shown.
- Line 145: it would be better if word “wavelength” is added after “450 nm”.

Results and discussion:  well described
- There is an interesting pattern from figure 2A  in the B-carotene reduction and content between 18-21 days which might be important to be explained

Table and figure:
- The data should be better if are shown in 3 decimal
- Consistency in writing ß in figure 2
- The statistical analysis of the differences in carotene and lycopene content during the storage is important to be shown (whether its in a different table or adding the error bar in figure 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: CHARACTERIZATION OF SPRAY-DRIED GAC ARIL EXTRACT AND ESTIMATED SHELF LIFE OF Β-CAROTENE AND LYCOPENE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript presents one of the interesting properties of gac fruit. However, the clarity and sentence structure of the manuscript require improvement. For example, the second sentence in the introduction requires rephrasing for more clarity.
Experimental design: The authors should provide values for the parameters analyzed before initial drying. This should be included in Table 1. Alternatively, they can explain why the data were not included in the report (or the experimental design).
Validity of the findings: 1. The values of the β-carotene found in this study are lower (Kindly check https://doi.org/10.1016/j.anres.2016.04.003). The authors should explain the possible scientific-based reasons for their results.

2. Make more comparison of the values obtained in your study with those obtained by other researchers for similar studies

3. Improve your sentence structure and grammatical correctness. For example, in line 252 and 253, where you stated that "...β-carotene and lycopene reduced from 1.403±0.019 and 0.225±0.004 to 0.117±0.014 and 0.033±0.004 mg/g sample after kept for 42 and 24 days (data not shown)", you need to add the word "respectively" after the sentence; except if stated otherwise.
Additional comments: Major revision is required as highlighted


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: CHARACTERIZATION OF SPRAY-DRIED GAC ARIL EXTRACT AND ESTIMATED SHELF LIFE OF Β-CAROTENE AND LYCOPENE
Review round: 2
Reviewer: 1

Output:
Basic reporting: It has been improved
Experimental design: Well described
Validity of the findings: well described
Additional comments: General comments:
All comments from the previous review have been addressed well, thank you. However, there are some minor comments:
RESULTS:
- Line 221-222: it is stated “This study showed no effect by spray-drying temperature on the water solubility of spray-dried aril powder”. In Table 1, water solubility index shows temperature 160 C had a higher WSI which is shown by statistical difference.Please check and confirm.
- Line 229-230: based on Table 2, all colour parameters are significantly affected. But it is written “....except for lightness”. Please check and confirm.
- Line 233-234: “the decreased in L…”from table 2, there is no difference in L, please confirm what this sentence means.
Line 235: delete (.)

DISCUSSION
- Line 272-273: delete  “and, in this study, the Custer DextrinTM.”
- Line 284: add “respectively” at the end of the sentence.
- Line 292, it is mentioned that differences in drying agents influence the reduction of carotenoid. But, in - line 299, it is stated that this study confirmed that the reduction occurred regardless the type of the wall material used. Does this mean that the result obtained in contrast with the other papers? If so, please state clearly. Or, if not, please make it more clear.


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: CHARACTERIZATION OF SPRAY-DRIED GAC ARIL EXTRACT AND ESTIMATED SHELF LIFE OF Β-CAROTENE AND LYCOPENE
Review round: 2
Reviewer: 2

Output:
Basic reporting: Satisfactory
Experimental design: Satisfactory
Validity of the findings: Satisfactory revisions
Additional comments: Satisfactory revisions


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: CHARACTERIZATION OF SPRAY-DRIED GAC ARIL EXTRACT AND ESTIMATED SHELF LIFE OF Β-CAROTENE AND LYCOPENE
Review round: 3
Reviewer: 1

Output:
Basic reporting: Satisfy
Experimental design: Satisfy
Validity of the findings: Satisfy
Additional comments: Thank you very much for your revisions and congratulations!


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: USING DEMOGRAPHIC DATA TO UNDERSTAND THE DISTRIBUTION OF H1N1 AND COVID-19 PANDEMICS CASES AMONG FEDERAL ENTITIES AND MUNICIPALITIES OF MEXICO
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. I am not sure of the referencing style used, where in most cases a URL has been provided and a year given in the manuscript. It should be the author/organization and year

2. Review of the risk factors of the pandemic not sufficient

3. The title does not reflect the analysis conducted

4. The current paper would fit better into a narrative style for COVID-19, otherwise more detailed analysis needed
Experimental design: 1.	It's not clear what the first objective is when the author says analyse. This should be phrased more clearly.

2.	Line 125, what do cc and d mean ( prefixes in COVID-19) ccCOVID-19, dCOVID-19. Did the authors mean comparisons of COVID-19 cases and deaths, then write in full before abbreviating. The same applies to ccH1N1 and dH1N1

3.	Line 136, how were the factors arrived at? Many factors would have been considered, why the specific ones listed and not others. In literature, there have been many risk factors of COVID-19 including for severe cases and deaths.

4.	 What does (hab.)  mean? If it’s the unit of measurement, write in full then in brackets the abbreviation. The same applies to the other units of measurements used in the manuscript

5.	The methods are not clear. Please tabulate all the factors considered after clearly indicating the rationale for their inclusions. Why was correlation chosen as the measure of association? Why did the authors not use regression analysis where they would also control for cofounders and spatial autocorrelation?

6.	The title Mapping the clustering of people to further understand the spread of viral infections is not appropriate given the evidence presented. There was no clustering analysis presented or any evidence on the spread of viral infections.

7.	The current analysis has looked only into correlations between the pandemics and randomly chosen factors among many risk factors. The title should be paraphrased to reflect the analysis done. Or add more analysis such as clustering to substantiate the title.

8.	Line 157-181, these are not results but country context details. Add a section at the beginning of the methods called country context. Some maps would help visualize these data in the country context section.

9.	Line 183 to 202, the details presented are mainly describing the pandemic with few results or correlation. Would the authors consider this being a description of the two pandemics as opposed to analysis paper? The details presented would fit better into a paper describing how the COVID-19 ( exclude H1N1) unravelled in Mexico and the control measures the government has been putting into place.

10.	 Further, why investigate the correlation between deaths and cases of the same pandemic? A death will be a case in the first instance. Further, there are different riks factors of deaths and different risk factors of the cases, therefore the variables should have been separated to those that are associated with a death and those associated with a case. These can be mined from the literature of the two pandemics. The same applies to the COVID-19 description

11.	The authors should note that correlation does not refer to causation. For example, in line 247, the authors are implying causation which is not quite right

12.	The cases presented in the maps should also be corrected for the population. The more populate areas will have cases, therefore dividing by the population gives a better measure per geographical unit
Validity of the findings: The discussion, methods and results can be combined into a narrative/descriptive paper describing how the COVID-19 ( exclude H1N1) unravelled in Mexico and the control measures the government has been putting into place.
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: USING DEMOGRAPHIC DATA TO UNDERSTAND THE DISTRIBUTION OF H1N1 AND COVID-19 PANDEMICS CASES AMONG FEDERAL ENTITIES AND MUNICIPALITIES OF MEXICO
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is written in clear, unambiguous, and technically correct language.
The introduction and background revealed the impact of the disease and the relevance of the analysis performed. some edits are suggested.

I find confusing when the authors describe percentages using “% X/hab.” (As done for the variables ‘%ccH1N1/hab.”, “dH1N1/hab.”, “%ccCOVID-19/hab.” and “%dCOVID-19/hab.”) I would remove the "/hab." from the names it since it could be misinterpreted and you are already describing that you are using a percentage of the total population (i.e. using the name “%ccCOVID-19” instead of "%ccCOVID-19/hab.").

Raw data has duplicated columns per sheet, non descriptive column names, and not well labeled for the disease outcomes analyzed (it doesn't specify which table is for which disease).  Providing a data dictionary and cleaning it a bit more is suggested.
Experimental design: The aims were well established, and the presented results describe well the relationship between population density, and case counts and deaths.

I don’t see the point of getting a correlation between the case counts and deaths since this just describes the same as disease specific mortality, which I think it would be more interesting to look at and can provide more succinct and complete information. I suggest the authors to use disease specific mortality to describe their results on the lines 213-239 and include it in the tables and thematic maps.
Validity of the findings: Underlying data have been provided and conclusions were limited to supporting results.
Something that is not mentioned and I think is very relevant to the conclusion on the geographical scale, is that when you've from the federal level to municipal, you are effectively increasing your sample size for quite a lot (from 32 to 2455). Smaller p-values are expected when you have a larger sample size and the null hypothesis is true.I would suggest to add something about this to the manuscript.
Additional comments: No additional comments


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

Input:
Title: USING DEMOGRAPHIC DATA TO UNDERSTAND THE DISTRIBUTION OF H1N1 AND COVID-19 PANDEMICS CASES AMONG FEDERAL ENTITIES AND MUNICIPALITIES OF MEXICO
Review round: 2
Reviewer: 1

Output:
Basic reporting: I found some gramatical errors in lines:
31 -  Should be “to collect…”, instead of 'to collected..."
451 - Did you meant “viral diseases” instead “viral deceases”?

For some sentences I understand what you are trying to say, but I would suggest to try to improve it to ensure that the audience clearly understand, including sentences from lines:
123-124 “There are no standardized medical therapeutics or vaccines against COVID-19 are just coming out and there is probably no pre-existing immunity in the population”, if you meant to mention about the newly developed vaccines or the treatments, I think you need to paraphrase it better.
154 - I suggest to add the last part as a third objective, i.e. “c)  investigate the correlation between the pandemic…”
284 -  To avoid any confusion, I suggest to change ‘crude mortality rate’ for ’cause specific mortality’. Crude mortality rate could be interpreted of deaths by any cause.
454 - "The local (municipality) scale that consisted of 2455 data the only 32 data of federal scale showed typically “smaller p-values” decreasing the uncertainty of the analysis" This sentence is a bit unambiguous please be more specific regarding what you mean with "typical smaller p-values" part.

The raw data looks great
Experimental design: For the regression, I suggest to also provide more details such as the p-value and/or confidence intervals for the coefficients, which would provide a better idea of the uncertainty of your estimations.

Last time I suggested you to use disease specific mortality, which I really meant to say disease fatality (deaths/cases) I apologize for my mistake, but I want to mention it because I think it should be very quick to calculate and could provide you interesting information for your tables and thematic maps.
Validity of the findings: My main concern regarding the regression model used, is that you didn’t explored or even mentioned anything about spatial autocorrelation. Remember that one of the assumptions of regression is independence, which is violated when you have autocorrelated samples.
So maybe consider expanding on the discussion about spatial autocorrelation mentioning it as a limitation or including a random effect for the municipal level model to account for this.
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: USING DEMOGRAPHIC DATA TO UNDERSTAND THE DISTRIBUTION OF H1N1 AND COVID-19 PANDEMICS CASES AMONG FEDERAL ENTITIES AND MUNICIPALITIES OF MEXICO
Review round: 3
Reviewer: 1

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


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

Input:
Title: USING DEMOGRAPHIC DATA TO UNDERSTAND THE DISTRIBUTION OF H1N1 AND COVID-19 PANDEMICS CASES AMONG FEDERAL ENTITIES AND MUNICIPALITIES OF MEXICO
Review round: 3
Reviewer: 2

Output:
Basic reporting: The grammar and structure of the text has improved, well done
Experimental design: The authors made several changes after the suggestions we did, great work!
Validity of the findings: Everything looks ok.
Additional comments: Thanks for addressing the comments, I am satisfied with the improvements you have made to the manuscript.


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

Input:
Title: THE EFFECT OF CARBON MONOXIDE ON MEIOTIC MATURATION OF PORCINE OOCYTES
Review round: 1
Reviewer: 1

Output:
Basic reporting: In general, it is a well written manuscript; however, the introduction section lacks recent references, they did not include articles from 2019 or 2020 I suggest including some recent articles with the use of antioxidants during in vitro culture.
Experimental design: Line 79, is it TCM-199 media instead of M199?
Provide the number of evaluated/used oocytes per experiment or total in the experimental design
Lines 91-92, why these concentrations were chosen?

Statistical analysis section
All figures only present the mean + SEM, not mean +- SEM, please modify

Discussion should include recent studies,  2019 and 2020 references are lacking
Validity of the findings: Findings are interesting; however, authors should indicate at which concentration CO inhibits maturation or reduces ROS. Conclusion is speculative and should only indicate the findings
Additional comments: No additional comments


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

Input:
Title: THE EFFECT OF CARBON MONOXIDE ON MEIOTIC MATURATION OF PORCINE OOCYTES
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: 1.	The deduction that is given from line 63 is interesting, however, it would be important to point out if there is any previous study carried out in oocytes, even in other species, if not, mentioning the absence of other studies would give greater relevance to this work.
2.	It would be important to check if there is more recent literature, in particular lines 54-57 and 66.
3. In lines 107-108, verify the meaning of RT-PCR, it would be "Reverse Transcriptase Polymerase Chain Reaction". Which would coincide with the transcription of RNA to cDNA that was carried out in the methodology.
4. On line 86, put CO2 with the subscript 2.

Overall, the work is very well done and promising. It leads to further studies.


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

Input:
Title: THE EFFECT OF CARBON MONOXIDE ON MEIOTIC MATURATION OF PORCINE OOCYTES
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Since the authors are stating that CO inhibits meiotic maturation, it would have been interesting to know whether or not this inhibition is reversible.
Since the authors are suggesting that CO may impact signaling pathways, it would have been interesting to know whether or not cumulus cels are contributing to this inhibition.


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

Input:
Title: THE EFFECTS OF PARENTAL AGE DIFFERENCE ON THE OFFSPRING SEX AND FITNESS OF EUROPEAN BLACKBIRDS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Uploaded review
Experimental design: Uploaded review
Validity of the findings: Uploaded review
Additional comments: Uploaded review


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

Input:
Title: THE EFFECTS OF PARENTAL AGE DIFFERENCE ON THE OFFSPRING SEX AND FITNESS OF EUROPEAN BLACKBIRDS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Thhis is an interesting paper described, sex ration in European blackbirds in relation to various parameters. Manuscript is whriten well, used clear and understanding English.
In Abstract section related to the methods is not related to the really used methods in the study.  Authors suggested long-term studies, therefore analyses are based on data from only four breeding seasons. In this part authors should give information about methods related tho the main goals of the paper.
Experimental design: The material seems to be well documented but, after carefully reading, I have some questions that need clarification.
My main doubts are related to the material used in this manuscript. Lack is information that part of analysed data were published in earlier paper: Nowacki P, Piliczewski P, Rek T, Kiriaka B. 2016. Secondary sex ratio of nestlings of the blackbird (Turdus merula) Secondary sex. Acta Biologica 23:69–74. DOI: 522 10.18276/ab.2016.23-06. In both papers was used data from the same population and one period; breeding seasons 2006-2007.
In Methods section some information should be added. How hatching date was determined? In which way fathers of the nestlings were determined? Authors stated that “relative chick mass on the day of ringing as a measure of hatching order”. It needs more details, criticism and discussion of used method.
Validity of the findings: The fidings are interesting and worth for publication. The conclusions are related to the aims and questions, and are supported by the results. However  in the Discussion section authors indicated that they did not find that laying date affected the sex ratio in the brood. But no such analyses exist in this manuscript.
Additional comments: No additional comments


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

Input:
Title: THE EFFECTS OF PARENTAL AGE DIFFERENCE ON THE OFFSPRING SEX AND FITNESS OF EUROPEAN BLACKBIRDS
Review round: 2
Reviewer: 1

Output:
Basic reporting: This article is improved much. The manuscript is written in clear English and text is technically correct. The article conforms to professional standards. This is an interesting paper described, sex ration in European blackbirds in relation to various parameters.
The structure of the article contains all standard sections.
Abstract section now is improved and is related to the used methods and obtained results.  The introduction is sufficient,  based on prior literature and knowledge related to the main topic of the article.
Experimental design: The method section is improved. At this moment authors cleary define goals and the research question. Methods are described with sufficient details. My questions related to the determination of the hatching, fathers of the nestlings, relative chick mass are explained.
Validity of the findings: The data are properly analysed using appropriate statistical tools. Conclusions are besed on results, well stated, related to original research question.
The manuscript fullfils gap knowledge related to the determination of sex ratio in nestlings and  heritable traits of the parents. The manuscript is  improved  and in this form is worth for publication.
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: TWO PREDICTED MODELS BASED ON CERNAS AND IMMUNE CELLS IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The contribution of this paper, in terms of novelty and performance improvement, is not enough.
Experimental design: No comment.
Validity of the findings: The introduction and discussion of variety of the algorithm principles used in this study and the responding results were insufficient.
Additional comments: This study constructed two cox regression models based on crucial ceRNAs and immune cells to predict prognosis in LUAD. This is a meaningful study, but there are several issues that the authors should address.
1. Two cox regression models based on crucial ceRNAs and immune cells respectively were constructed in this study, however, what the correlation of these two regression models were stated insufficient. In short, the contribution of this paper, in terms of novelty and performance improvement, is not enough.
2. In this study, ESTIMATE and CIBERSORT were mentioned to evaluate or estimate the proportion of stromal and immune cells, what’s the different between these this two methods, and why these two methods are both chosen should be clarified.
3. In addition, in the ‘Multiple databases validation’ part, UCLAN, GEPIA and TIMER (line 181) are also used on different databases; and in line 194, ssGSEA also as an algorithm for calculating the abundance of immune cells was used, I doubted why so many different immune cell infiltration related abundance-calculating methods were used on different datasets? If the authors want to compare the effectiveness of different algorithms, the results of different algorithms on the same dataset should be given, or the same algorithm is compared on different datasets.
A wide range of different datasets were used in this manuscript, at the same time, a variety of methods were used to calculate the abundance of immune cells such as CIBERSORT, ssGSEA, UCLAN, GEPIA and TIMER, how and why these different abundance calculating methods were selected was not stated. A sufficient discussion of the advantages and disadvantages of each method and why they are chosen should be provided.
4. For Figure 3 and Figure 7, the content in the figures should be described in detail, such as the meaning of the shapes, colors and size of the node, and so on.
5. This article uses many algorithms like CIBERSORT to calculate the abundance of immune cells, but the introduction and discussion of the algorithm principle and some necessary details of parameter setting for all the applied methods were missing.
6. A variety of datasets were used in the manuscript like mRNA, lncRNA, miRNA, for each dataset, however, the details including the number of samples and RNAs, information of patients and so on are missing.


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: TWO PREDICTED MODELS BASED ON CERNAS AND IMMUNE CELLS IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Grammatical error in line 28, 169.  spell check for DESeq in line 69
Figure legends need to be more elaborate
Experimental design: na
Validity of the findings: The authors have efficiently used the published dataset and tried to eludeidate a bunch of biological questions related to LUAD.

In performing the differential expression between normal and tumor
      A. Did the authors compared their results to the results from 3 previous studies ; https://doi.org/10.1186/s12935-020-01295-8, https://doi.org/10.3892/ol.2018.9336, https://doi.org/10.3892/ijo.2016.3716, https://doi.org/10.1111/jcmm.15778
      B. Looking at the heat maps of the DEG in fig1c - the 2 groups are not segregateing very well based on the clustering. If the genes were significantly different, I would expect clustering to separate them out.
      C. A fold change of >|1| seems a bit lenient especially considering the fact effect sizes in tumor vs normal
      D. For the dataset used in the first set- were any other co variants taken into account? Especially doing the cell type estimation

2. It is not clear what motivated the authors to look into the smoking status in nomograph predictions?
3. Is it possible for the authors to show some kind of clustering for the high vs. low risk groups transcriptomic profiles overlayed with any other important covariate?

4. More information is needed in the figure legends to make it more interpretable.

5. Also for the validation dataset, which seems to be a set of samples with different mutations, how was the dates used or conditioned for these mutations.

6. Now that there are tremendous amount of single cell transcriptomic data available, can the authors validate/look up some findings in any public datasets if available?

7. In fig 5E- except for a subset of samples- the distribution of the >20 and <20 smoking status look very similar. Did the authors look into what that subset is which is driving the most differences?
Additional comments: The authors in this manuscript used published dataset to identify differentially expressed mRNA,miRNA,lncRNA between normal tissue and tumor samples which were then used to construct a regulatory network of interactions. They further predicted patients prognostic values based on these findings. They also used CIBERSORT to estimate the immune cell types differences in normal vs. tumor and eventually integration into the multivariate model. They found the hub ceRNA to be associated with stemless and the microenvironment of which the key members were validated in another independent published dataset.


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: TWO PREDICTED MODELS BASED ON CERNAS AND IMMUNE CELLS IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: NA
Experimental design: NA
Validity of the findings: NA
Additional comments: The authors have addressed the concerns.


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

Input:
Title: IMMUNOINFORMATIC APPROACH TO DESIGN A MULTIEPITOPE VACCINE TARGETING NON-MUTATIONAL HOTSPOT REGIONS OF STRUCTURAL AND NON-STRUCTURAL PROTEINS OF THE SARS COV2
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript requires thorough proof-reading to remove errors.
Experimental design: The author has carried out Immunoinformatic approach to design a chimeric multi-epitope vaccine against non-mutational hotspot region of structural and non-structural proteins of the SARS CoV2, Notwithstanding the study fails to have a novel approach and steady conclusion.
Validity of the findings: The results are not novel at any point and add no further information to the already known research in this field. The authors must make the needful incorporations before considering to communicate the article further.

•	The 3D structure of construct is modeled on homology principle which even if just prediction very wrong as the construct sequence is showing very less sequence homology to the available experimental structures. The authors should use ab initio principle instead.
•	The docking studies must be cross validated by another protein-peptide server to assure correctness of the binding mode with the receptor.
•	Further, MMPBSA binding free energy calculations must be performed on the MD trajectories to validate the docking predictions.
Additional comments: Some serious questions need to be answered before considering this work for publication. Based on my review, I concluded the rationale of designing a vaccine when already licensed vaccines are available this work is very weak for publication.
•	Why this study is performed when already licensed vaccine for SARS-COV2 is already available? This comment must be answered very clearly in the introduction section and strong rationale should be provided if any?
•	How this vaccine construct is different from that many reported in the literature?


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: IMMUNOINFORMATIC APPROACH TO DESIGN A MULTIEPITOPE VACCINE TARGETING NON-MUTATIONAL HOTSPOT REGIONS OF STRUCTURAL AND NON-STRUCTURAL PROTEINS OF THE SARS COV2
Review round: 1
Reviewer: 2

Output:
Basic reporting: Abstract: Need to give more reasons about the significance of your study and how it will leads to the formation of novel vaccine.

The manuscript is clear and professional english is used. The literature is well discussed.

In Paragraph 2.1, Protein sequence collection: Mention  more information about the retrieval of protein sequence. For instance, the source, organisms name, incubation period, site etc.

In Paragraph, 2.12, Give more dara on which sequences of human microbiome was taken into consideration.
Experimental design: The experimental data is discussed very well.

In fig.3, Meet to add more information regarding what is the significance of red, yellow and green colour spirals shows.
Validity of the findings: The validity of findings is perfectly mentioned in the manuscript.
Additional comments: Need to check the palgriasm of the whole manuscript. Rest of all findings are well discussed.


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

Input:
Title: IMMUNOINFORMATIC APPROACH TO DESIGN A MULTIEPITOPE VACCINE TARGETING NON-MUTATIONAL HOTSPOT REGIONS OF STRUCTURAL AND NON-STRUCTURAL PROTEINS OF THE SARS COV2
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1- Although the manuscript is fairly easy to read and understandable, grammatical errors can be found throughout the paper (just a few examples have been highlighted in blue ). I might suggest the authors check the manuscript for all language errors and inconsistencies to reach the international audience.

2- The references are properly formatted. However, it seems references are lacking in certain sections (e.g., lines 90-95, line 180; no reference for PATCHDOCK server, line 321; should change to (Tiwari 2020a, Tiwari 2020b) as opposed to (Tiwari 2020a) (Tiwari 2020b), etc). Please check draft and add references where necessary throughout the manuscript.

3- All tables, figures, and data have been provided in good quality and scientific clarity. In particular, I would like to mention Figure 1 is very well-designed, and much-appreciated as it clarifies your approach towards chimeric protein engineering.

4- The research question is clear, and has been backed up by sufficient evidence in the main text. However, the background section of the abstract needs more explanation. It is vital to describe the rationale behind the employed in silico approach more precisely.

5- Lines 36-39: Moving such points to the methods section may improve the integrity of the abstract. The authors can start the results section by mentioning a multi-epitope vaccine was built. The results section of the abstract could as well include more precise quantitative results.

6- Lines 120-121, 137-144: The long list of HLAs is appreciated, as it increases vaccine coverage, and shows the precise reporting of study. However, it would be easier on the eye if only tabulated.
Experimental design: 7- It is appreciated that, the knowledge gap has been identified properly in the title and throughout the paper as "targeting non-mutational sequences of SARS-CoV2".

8- Lines 70-73: I might add the authors mention a high affinity to ACE2 for mutational regions of the SARS-CoV2 S protein. This seemingly contradicts their rationale behind selecting non-mutational regions, in particular from the surface glycoprotein (NCBI Protein code: YP_009724390.1). I might suggest such points are include in the discussion section, as possible limitations of vaccinology against COVID-19.

9- I would like to appreciate the authors' work for Sections 2.8, 2-12, and 2.13, as these sections highlighted "cross-reactivity" and "mutation" an a vital consideration in the in silico approach towards combat against viral infections. I might encourage the authors highlight points related to these sections further in the discussion section.

10- Section 2.11: The use of up-to-date software like Desmond is appreciated, as it is a widely-accepted approach to MDS. I might suggest you explain this section with further MD settings and details. Moreover, it would be great if you could perform MM-PBSA analyses, or similar binding analyses during the MD run (see: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4487606/). If MM-PBSA analyses are not possible, you can extract MD trajectory frames every 1-2ns, and perform binding analysis of the TLR-Vaccine complex by the PRODIGY server (https://wenmr.science.uu.nl/prodigy/) or similar tools. This gives your binding analysis much more validity and significance.

11- As the methodology stands now, the methods used in the present study are valid and robust. However, the current standard of in silico vaccine research indicates the need for more explorations, as have been used by many prominent studies (just to name a few, https://www.frontiersin.org/articles/10.3389/fimmu.2020.01784/full, https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0244176#, https://www.frontiersin.org/articles/10.3389/fimmu.2020.02074/full).

It is advised that the methodology is refined further in certain areas:
I.	The present protein structure quality assessment method should be extended. The 3 region Ramachandran plot has been shown to be beneficial, yet inadequate for the purpose (see: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3061398/).
Please assess the designed protein quality further by ProSA tool (https://prosa.services.came.sbg.ac.at/prosa.php) or QMEAN server (https://swissmodel.expasy.org/qmean/), and provide a Z-score based validation for your protein structure as it is much more reliable than solely using a Ramachandran plot (see: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3031035/).

II.	 To determine the conserved regions of obtained FASTA formats, you can perform multiple alignment by online servers or BioEdit software and only select the conserved region across all proposed PubMed Protein FASTA formats.

III.	You can submit your PDB structure to GalaxyRefine server (http://galaxy.seoklab.org/cgi-bin/submit.cgi?type=REFINE), and provide a comparative Ramachandran plot to indicate the protein structure has been properly refined.

IV.	Please explain why your sequence does not start with "M". This may cause issues with real-world vaccine expression (see: https://pubmed.ncbi.nlm.nih.gov/20970746/). You may use SnapGene software (https://www.snapgene.com/) to conduct in silico cloning, and determine if the vaccine can be experimentally synthesized in a specific organism (e.g., E. coli).
Validity of the findings: 12- The results are very valid, but may be enhanced, in particular in the area of protein quality validation. I might suggest the authors use suggested servers and add their findings to enrich the paper.
Additional comments: 13- Generally, the present research is very well-conducted and can contribute to the field of in silico vaccine research. However, the methodology should be at least in part extended, in particular in the area of protein structure refinement and validation. Language and the structure of the manuscript, in particular the abstract, should be improved.

Note: All specified lines which may require enhancements have also been highlighted in yellow in the attached PDF. Also, there are many positive attributes to this work, which have been highlighted in green.


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: IMMUNOINFORMATIC APPROACH TO DESIGN A MULTIEPITOPE VACCINE TARGETING NON-MUTATIONAL HOTSPOT REGIONS OF STRUCTURAL AND NON-STRUCTURAL PROTEINS OF THE SARS COV2
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: English language has been improved significantly. Also, the authors have refined their methodology to match state-of-art techniques, bringing remarkable significance to their findings. Finally, the work adds new visions into vaccine development, in particular, because the authors have taken viral mutations and cross-reactivity into account, an aspect often overlooked in the existing in silico literature. I would like to recommend acceptance and congratulate authors for their novel work.


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: SPATIOTEMPORAL DYNAMICS OF HABITAT SUITABILITY FOR THE ETHIOPIAN STAPLE CROP, ERAGROSTIS TEF (TEFF), UNDER CHANGING CLIMATE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The agricultural systems in African countries are mostly rain-fed, and it is important to predict the crop yield in Africa. Therefore, the topic of the paper is interesting and the background of the study is well addressed. But the paper is poorly prepared. There is no workflow and the result, discussion, and conclusion sections are confusing. It is very difficult to understand what is new in this paper. Authors should read more academic papers and try to learn how to organize a scientific paper.
Experimental design: There is no workflow, and the method is not well described.
Validity of the findings: It seems there are some experiment results relates to the objectives in the result section, but there is no further analysis, and the conclusion section does not relate to the objectives listed in the introduction section.
Additional comments: Major concerns:
There is no workflow included in the paper. So that the experiment design is not clear. The authors should clearly show how they address the three objectives. I think the methodology section should be re-structured.
Line 253: Is Jackknifing a toolkit used for variable importance analysis? The authors should explain the tool kit in the method section.
Line 294: Is MaxEnt used to predict the distribution of teff? if yes, please analyze the teff distribution maps predicted by MaxEnt.
The discussion section should compare the result in the paper with other existing studies, show the advantages, limitations, and uncertainties of the paper.
line 416; In the conclusion section, I did not see anything related to the three objectives listed in the introduction section.

Minor concerns:
line 84: "have increased 0.85 ° from 1880 to 2012 0.85 ° C"
line 141-142: "genetic algorithm for the production of rules (GARP), BIOCLIM (the model widely fused tools to predict current and future species distribution of response)"
What are the number is the x-axis of Figure mean?


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: SPATIOTEMPORAL DYNAMICS OF HABITAT SUITABILITY FOR THE ETHIOPIAN STAPLE CROP, ERAGROSTIS TEF (TEFF), UNDER CHANGING CLIMATE
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper is generally very interesting with great results. However, the flow of thought in the paper is poor. The introduction, for example, is poorly structured with several editorial errors. The sentence on lines 119 – 121 (see the attached annotated PDF document) suggests the information on Teff is over but all of a sudden, the authors (by the sentence on lines 122 - 125) again bring in climate issues (which were discussed earlier). My suggestion is that the authors rearrange the introduction to ensure a smooth flow of thought. I would advise that the authors finish talking about climate issues and how they affect food production before introducing Teff. Afterward it not suitable to bring in climate again in the introduction because it disrupts the flow of thoughts.

Also, the sentences on Lines 192 – 200 (under the subsection "Teff Distribution Data") should be removed entirely from the manuscript or moved to other sections like STUDY AREA or INTRODUCTION where they may best fit.

I also noticed the same back and forth movement of thoughts in the discussion section.

Finally, I think the abstract is too long and needs revision.
Experimental design: Very well written and well explained. However, the authors should address the the following comment:

The parameters used for running the MaxEnt model have been provided but no explanation is given. Did these parameters provide the best model performance, for example? It is important to tell the reader. I think the 75%/25% training/test split is more or less a rule of thumb so explanation is not necessary.
Validity of the findings: no comment
Additional comments: A few more comments:

1. How different is the information on cultivated area of Teff (Lines 114 & 115) from “Teff crops occupy 988,638.5 square kilometers” (Lines 130)? They appear confusing to the reader and should be revised if both are needed.

2. Stewart & Getachew (1962) which was used to support the sentence "It is estimated that Injera, made from teff, provides up to two-thirds of the food consumed by Ethiopians" is too outdated. The situation may have changed. I suggest the authors use a more current reference.

3. Please be consistent with your measurement scale. If you want to use , m^2, km^2, etc. then convert other values from ha to metric scale as well.

4. Did the authors preform any statistical significance testing? I ask because of a sentence like “The potential teff distribution under current projected climate change was significantly affected…”

5. Some headings have all words capitalized while others do not. Ensure consistency.

6. Change bio1, bio2, … to  Bio1, Bio2, … so that they are consistent with the rest of the text.


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

Input:
Title: SPATIOTEMPORAL DYNAMICS OF HABITAT SUITABILITY FOR THE ETHIOPIAN STAPLE CROP, ERAGROSTIS TEF (TEFF), UNDER CHANGING CLIMATE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The paper has been improved significantly and I could better understand the work with the revised version. Then, I still have some concerns.
Experimental design: it's fine.
Validity of the findings: The result need more explanations, please see the Comment in the "General comment".
Additional comments: 1. I suggest to use a figure to show the workflow of this paper, for example, it seems there are two steps for this paper (1. variables importance estimation and 2. teff distribution prediction based on multiple climate conditions), the workflow figure should show inputs and outputs of each step and then link these several steps together. This workflow figure could make it clear to the readers how the authors have designed their experiments. In addition, its new section should be added as the "workflow of the study" to describe the workflow figure.

2. Line 331~333 and 364~369，can you explain why your results are different from the existing studies? As they have concluded that precipitation in the wet season affected spatial of teff but your conclusion is the lowest temperature in the coldest season affects teff distribution the most, so can you any principle which could support your conclusion? I think this is important as the teff prediction result is highly dependent on the high contribution factors.

3. Line 386~403, this paragraph is not related to any experiment results, it seems it is better to provide this background information in the "Introduction section".

4. I suggest further discuss the possible effects of climate data uncertainty (particularly the uncertainty of future climate data) on your teff prediction results.

5. The conclusions section could be further improved. I suggest summarizing the conclusions as (1) conclusion of variables importance, (2) conclusion of predicted teff distributions, and (3) effects of climate on teff distributions based on your 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: SPATIOTEMPORAL DYNAMICS OF HABITAT SUITABILITY FOR THE ETHIOPIAN STAPLE CROP, ERAGROSTIS TEF (TEFF), UNDER CHANGING CLIMATE
Review round: 3
Reviewer: 1

Output:
Basic reporting: All my concerns have been addressed and I am satisfied with the revision.
Experimental design: It's fine.
Validity of the findings: It's OK.
Additional comments: All my concerns have been addressed and I am satisfied with the 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: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Written text

The manuscript is well-written in general, but a fine tuning in the written English should improve some details in the text. I made some suggestions over the manuscript with track changes. I don’t believe it needs a native speaker or professional editing for the language revision.

Also, please revise typos and missing commas. Example "et al.," and not "et al." Revise terms in italics, such as “i.e.,”.  Revise values and statistics regarding empty spaces between values and symbols.

Table
For Table 1, herkogamy should be written in full. There is no information on which side is for which population. Besides, Table 1 is referred in the text when authors discuss on progenies and lineages (ie, basically the first four lines or the other lines, but separately). To me, it is confusing. It would be good if in the Table, not only the population information is cited, but also the progeny or lineage "test" aspect of the comparisons.

Figure in general
Figures represent well the results. However, much improvement is needed. In my opinion:
-	there should be a standard way to refer to single panes in the figures (use of letter A, B, etc across figures should be similar).
-	Colored bars in grey can be spared for simplicity and for less ink use.
-	Make sure to produce graphs with similar font types and sizes. Some axes are represented with values in bold and normal font type, others are in bold and rotated by 90 degrees, for instance.
-	P values shown are not equal to 0. Thus, they should be reported as for example P < 0.001 and not P = 0.00.
-	Non-significant slopes (lines) should actually not be plotted in my understanding (see Good, P. I., & Hardin, J. W. (2012). Common errors in statistics (and how to avoid them). John Wiley & Sons.). In case authors find it necessary, maybe distinguish them from significant correlation lines making them dashed or in lighter color.
Specific figures

Fig. 1: it seems that the tip of error bars are incomplete (only half) in some cases. Vertical bars are at some points separated by white empty spaces and other times not.

Fig 2: and its caption should be improved. Labels for areas (population) should be centered and reduced in font size. X Axis should better represent the values for the histogram bars (the way it is now is a bit misleading). Besides, the caption should clarify that for each population there are two graphs (boxplot and histogram). At first glance the separate plots seem to be isolated. Color for bars and boxplots could be spared.

Fig. 3 and 4: specify which correlation estimation used in caption (ie. Pearson’s correlation…), consider including the slope/R2 in the significant regression lines. Letters A-H were not mentioned in the caption. In Fig 3, dots may be set as translucent to allow overlaying points to be seen better.
Experimental design: •	The submission should clearly define the research question, which must be relevant and meaningful. The knowledge gap being investigated should be identified, and statements should be made as to how the study contributes to filling that gap.

The final paragraph of the Introduction section sounds to me more like a Methods (ie, species and area description), and as such this info should be merged with the info in Methods. Additionally, it would be perhaps meaningful to formulate a better Introduction end, by stating the main gap in the knowledge and the objective of the study.

•	Methods should be described with sufficient information to be reproducible by another investigator.

A statistical analyses section should be created and more detailed in my view. It is fragmented between sections, and thus a bit repetitive (with regards to software used). Moreover, I made some comments in the annotated manuscript
Validity of the findings: •	Replication experiments are encouraged (provided the rationale for the replication, and how it adds value to the literature, is clearly described); however, we do not allow the ‘pointless’ repetition of well known, widely accepted results.

Authors could discuss on the presence of other populations or related species that might have been added to the story about the evolution of selfing in Datura. This would contribute to the Discussion section in my view.
Additional comments: CM and Map abbreviations are actually not always used across the text.

Comments and suggestions by line
line 20: "Nevertheless, despite the potential advantage of selfing, it may restrict gene flow within and among populations producing high levels of inbreeding between individuals and increasing homozygosity (Ho), and thus the likelihood of inbreeding depression (ẟ)"
In my opinion this long sentence could be reformulated and more clearly be contextualized. For example: Despite the potential transmission/reproductive advantage, selfing as a mating system may as well restrict gene flow within and among populations. Consequently, selfing might increment levels of inbreeding and homozygosity (Ho), and ultimately the likelihood of inbreeding depression (ẟ) in populations.

line 47: "(i.e. no herkogamy)"
Is it the same as nil herkogamy. Nil herkogamy was not defined in the manuscript.

line 230: "To estimate phenotypic variation in herkogamy for each population, we calculated the within- and between-individual variances. Individual plants were considered a random effect. The residual variance is accounted by the intra-plant variation. The significance of individuals was evaluated by a likelihood-ratio ꭓ2 test (LRT) between a model that includes individuals as random effect (function lme) and one that does not (LRT1; function gls) (Zuur et al., 2009). Analyses were conducted with nlme package (Pinheiro et al., 2017) in R software (R Development Core Team, 2008)."
It is not clear what this step is testing and why. At this point, I didnt understand why authors had to compare differences between individuals, but rather herkogamy vs. populations for instance. Since individuals are taken as random effects, how can they be tested for significance rather than being assigned as a fixed-effect predictor? Thus, it needs a better clarification.

line 240: "...glasshouse..."
Maybe describe the conditions of light/humidity/temperature

line 248: "Since r is a proportion with binomial distribution, correlation with herkogamy was calculated by a generalized linear model with a quasi-binomial distribution error (Crawly, 2013)."
I understand that the calculation of correlations using proportional data pose a mathematical problem compared to traditional continuous data correlations. I am not completely convinced, however, about the method used, since the use of quasibinomial distribution assumes a particular variance structure, but this was not mentioned in the methods throughly. The reference on Crawly (2013) doesn't seem to actually explain the use of quasi-binomial distribution on testing correlation. My suggestion would be to (1) better fundament the use of the method (i.e., linear model and distribution) or (2) use another distribution that clearly fits into 0-1 such as beta/binomial (for example, in R one might use the betareg package to run a beta regression).

line 307: "To estimate inbreeding depression (ẟ), we collected 150 fruits from different individual plants in each population"
How many plants and did they include the tagged ones from the measures above? If so, the proportion of those would be interesting to know.

line 381: "This suggests that the inbreeding history at each lineage not only depends on herkogamy; instead, purge of inbred individuals and/or biparental inbreeding may also come into play."
However, they missed to discuss why it was the case by suggesting possible explanations and references that tackle this issue

line 398: "On the other hand, the mating system of the population of Cañada de Moreno is predominantly selfing, and no association between selfing rates, herkogamy and inbreeding coefficients in the adult cohort were detected."
However in Table 1, we see that

line 432: "pollination likelihood by animals" what does this mean. Better formulation


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: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors report on a study of two populations of Datura inoxia in Mexico, aimed at quantifying variation in floral traits, mating patterns, and inbreeding among individual lineages within each population. These associations have been well studied in a range of systems, but as the authors point out, relatively few studies have assessed these associations within (rather than among) populations.

The authors have assembled a rather large dataset on flower morphology, outcrossing rates and inbreeding coefficients derived from microsatellite markers, and have also quantified inbreeding depression experimentally in the greenhouse. I found the data collection to be adequate, and the dataset to be appropriate for quantifying the focal variables.

Most of my comments relate to the presentation of the manuscript, which I found less than ideal. Most of this is language issues, and could thus be fixed by revising the text.
Experimental design: The data collection protocols appear adequate to me, although the presentation could be improved in some places as outlined in my line-by-line comments.

In the analyses, I suggest considering testing for differences between populations in ANCOVA-type analyses (y ~ herkogamy*population). In the results can then focus more on the contrast between the two populations, than on the statistical significance of patterns within each population.
Validity of the findings: I agree with most of the authors conclusions based on the study, but have suggested some areas where further elaboration could increase the impact of the paper.

The relationship between individual-level herkogamy and mating patters differs a bit from the expectations for among-population relationships. Among populations we except herkogamy-outcrossing relationships to have evolved in response to variation in, e.g., the reliability of pollination. Within populations, a correlation between herkogamy and outcrossing rate could relate to processes such as stigma clogging and related mechanisms of sexual interference, as originally discussed by Lloyd and Webb 1986. I suggest expanding a little on the specific mechanisms in play at the within vs. among-population level. A recent review of these ideas is given in Opedal 2018 IJPS.

Unpredictable pollination service is casually discussed as a potential source of (fluctuating) selection on mating systems, but it is not so clear whether this relates to daily fluctuations vs. variation within and among seasons. The link from fluctuating selection (at the population level) to maintenance of individual lineages with contrasting mating histories is not that clear to me. Selection for reduced herkogamy as a mechanism of reproductive assurance, sure, but why would not the population reach an ‘optimum’ herkogamy, with some phenotypic variation around it (due to plasticity etc.). At least these issues could be discussed more explicitly.

The data allows estimating (early-acting) inbreeding depression in the greenhouse, and also (potentially late-acting) inbreeding depression in the field (the comparison of F to Fe, as outlined by Ritland 1990). This could be made a more explicit part of the presentation, as it is currently unclear how often these two approaches give similar results.
Additional comments: Some suggestions for clarification of individual points and sentences:

Abstract
I found it a little confusing to read about ‘genetic variance in herkogamy’ in the Abstract, as this does not seem to be addressed

Introduction
Line 2: Plant mating system affect the demographic dynamic and genetic structure of populations, and thus their adaptive potentials.
Line 4: Mating systems often vary widely among populations of self-compatible species.
Line 9: Maybe start directly from: ‘The transition from…’
Line 14: Two main mechanisms are thought to favour the evolution of selfing
Line 15: ‘genomes’ -> gametes
Line 41: See also Opedal 2018 IJPS for a review of the expected functional relationships between herkogamy and mating patterns, and a review of the literature.
Line 43: Herkogamy is a complex trait determined by…
Line 53: There are a few more studies, see the review in Opedal 2018 IJPS
Line 57: Maybe change heterogeneous to stressful?
Line 74: What suggests that the variation in adaptive? Association with different pollination environments?
Line 77-78: Or selection on herkogamy which affects the mating system and generates an inbreeding history? Not sure correlated selection needs to be invoked here.

Methods
Line 81: ‘Study species and sites’?
Line 85: September, similar to other Datura species
Line 88: stigma receptivity
Line 88: from a few up to
Line 90: Flower traits … vary widely within and among populations
Line 92: forage for nectar
Line 97: Maybe ‘herkogamy, selfing rate, and inbreeding coefficients’ is more logical?
Line 99: ‘e.gr’ - > including
Line 101-102: ‘indicate substantial within-population variation’
Line 110-102: Please clarify the focus here – daily fluctuations or seasonal variation?
Line 116: Measured as, or calculated as?
Line 117: Mention how approach/reverse herkogamy translates into positive/negative values as recorded in this study.
Line 121: I don’t think there is any need for this statistical hypothesis test. Simply quantifying the variance components should suffice (possibly with confidence intervals).
Lines 132-136: If 20 seedlings per maternal family was analysed, no need to point out that 25 were initially collected.
Line 142: shown -> described. (And thank you for that!)
Line 152-160: I like the analysis of primary selfing rates!
Line 161: Why is biparental inbreeding not a part of the mating system?
Line 169-170: ‘larger differences’?
Line 172: Consider citing Allard et al. 1968 here, where the equivalent formulation s/(2-s) appears.
Line 174 and throughout: multilocus outcrossing rate
Line 177: Is the herkogamy-outcrossing relationship ‘just a correlation’, or a causal relationship?
Line 178: error distribution
Line 184: on -> in. Please specify greenhouse growth conditions.
Line 186: reproduction -> flowering
Line 188: maternal plant
Line 189: hand-fertilized -> hand-pollinated
Line 195: fitness components

Results
Line 205: Variation in herkogamy within and among populations
Line 206: At Cañada de Moreno, plants were on average approach herkogamous, but with substantial variation (mean = m range =).
Line 208 and throughout: Consider replacing ‘showed approach herkogamy’ with something like ‘were approach herkogamous’ or ‘exhibited approach herkogamy’. Reads better at least to me
Line 212: Herkogamy varied proportionally more among plants at Mapimí than at Cañada…
Lines 214-216: I suggest dropping this hypothesis test. The variance components speak for themselves.
Line 221: to -> towards
Line 229-230: Make the distinction between the greenhouse and population-genetic estimate of inbreeding depression clearer.
Line 232: Relationships between herkogamy and mating-system parameters
Line 233 Maybe focus more on the difference between populations than on the statistical significance within each. Consider even testing for differences in an ANCOVA-type analysis (y ~ herkogamy*population)
Line 240-241: So the take-home here is that herkogamy is unrelated to the breeding history of the population? This would be highly surprising at least to me

Discussion
The discussion overall flows better than e.g. the introduction.

Line 249-250: Sounds a little strange that it is the ‘association between the traits and the mating strategies’ that is favored. Perhaps rephrase to reflect that certain traits (like herkogamy) is likely to coevolve with the mating system, because it is a direct modifier of the efficiency of self and cross-pollination?
Line276: Maybe explain -> facilitate?
Line 297: Very inaccurate. Rewrite to reflect the relationships between additive variance, evolvability (mean-scaled additive variance) and heritability (variance-scaled additive variance). What Opedal et al. 2017 argue is that herkogamy is highly evolvable on a proportional scale, i.e. it’s easy to change herkogamy by e.g. 10%, compared to changing e.g. flower size by 10%. This is in part due to relatively weak constraints arising form the covariation between pistil length and stamen length in many systems. Note also the high evolvability estimate for D. stramonium presented in Opedal et al. 2017, could be cited in support of the present findings of substantial among-family variance.
Line 301: Point out that these percentages represent proportional variation (because the two populations differ in phenotypic variance, the percentages do not map directly to variation in units of mm).

Table 1: Why not present this as a correlation matrix with standard errors/support values on one of the diagonals?

Figure 1: Eliminate the x-axis labels, as mostly confusing?

Figure 2: Do the box plots show means or medians (normally medians)?

Figure 3: Why are linear trend lines shown if analyses were done with binomial GLMs?

Figure 4: The hypothesis test seems strange for the deterministic relationships shown in C and D.


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: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comments
Experimental design: The experimental design is generally standard and consistent with previous research, except for some potential flaws that may require concerns.

1. Estimation of the inbreeding depression may be biased. Inbreeding depression is estimated based on the artificial pollination treatment. It is quite likely that an individual with a higher selfing rate will also have a lower number of deleterious mutations in the genome, which means a higher fitness. Given there is a high variation in selfing rate, this correlation may be strong.

However, this correlation between selfing rate and fitness is not reflected in the artificial pollination treatment. Therefore, inbreeding depression estimated using the artificial pollination experiment will be different from the inbreeding depression when pollination happens naturally. This inconsistency affects the calculation of the primary selfing rate, as the seeds used for estimating selfing rate are collected from the natural population.

2. The accumulative inbreeding depression (δ) used to calculate the primary selfing rate (r) is problematic. In the equation r=s/(1-δ+δs), δ accounts for the selfed offspring died from the stage of fertilization to the stage of the measure of the selfing rate s in the offspring seedlings.

However, it is said the cumulative inbreeding depression has two components: mean seed set and seed mass. Why can seed mass be used as a component of fitness? Is there any evidence in Datura inoxia showing a linear relationship between seed mass and the probability for a seed to germinate to a seedling? If not, then seed mass will not play a role in reducing the selfed offspring at the stage of the measure of the selfing rate, and should not be included into the cumulative inbreeding depression.
Validity of the findings: Generally the resutls presented are good, but there is one suggestion:

1. It may be unnecessary to show a correlation between herkogamy and the equilibrium inbreeding coefficient F_e in Fig. 3. F_e=s/(2-s) is a theoretical prediction based on the selfing rate s. Therefore, given the correlation between the primary selfing rate r and herkogamy has been already shown, it seems redundant to show a correlation for F_e.
The same logic also applies to Fig. 4, where showing a correlation between F_e and the primary selfing rate r is of no value, as it is almost guaranteed by the calculation of F_e. Also, based on the equation F_e=s/(2-s) and s=(1-δ)r/(1-δr), it is clear F_e will not have a linear relationship with r.
Additional comments: Generally, the paper is good and a standard investigation on the relation between phenotype, selfing rate, inbreeding coeffcient and inbreeding depression. It may be interesting to have an explanation why inbreeding coeffcient is negative.


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: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: I believe the authors adequately accepted/answered the issues raised in my previous feedback. After the revisions accounting for the comments and suggestions of the reviewers (including myself), I consider that the manuscript is adequate for publication. The written text and illustrations are improved and to my opinion ready for publication.

Two minor details, however, I would still raise for fine tuning (regarding text format only):

1) line 127: "error SD". I believe authors refer to standard deviation error. This notation should be the same as for the other notations across the text, which were noted as "sd" (lower case). Figure 1 caption also uses SD (uppercase)

2) line 203: "Statistical analysis" is a subsection of "Material and Methods", and therefore not bold, but rather in italics.

Finally, I would make one final suggestion, but I leave it to the authors to decide since it is not a fundamental point. Since figures comparing the two populations are more clean now, maybe using a nice colour code could help distinguish the two populations more easily.
Experimental design: no comments
Validity of the findings: no comments
Additional comments: no 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: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 2
Reviewer: 2

Output:
Basic reporting: See below
Experimental design: See below
Validity of the findings: See below
Additional comments: The authors have done a good job revising the manuscript, which is clearly improved from the previous version. I have made only minor comments and suggestions on the attached file. The text could benefit from some final polishing. I congratulate the authors on an interesting study!


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

Input:
Title: MATING SYSTEM OF DATURA INOXIA: ASSOCIATION BETWEEN SELFING RATES AND HERKOGAMY WITHIN POPULATIONS
Review round: 2
Reviewer: 3

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


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

Input:
Title: CHARACTERISTIC CHANGES IN MALT, WORT, AND BEER PRODUCED FROM DIFFERENT NIGERIAN RICE VARIETIES AS INFLUENCED BY VARYING MALTING CONDITIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall quality: The subject of the paper is interesting and it fits well with the scope of PeerJ journal, and with conclusions of interest for the scientific community. Regarding the conception of the paper and the presented results, the paper can be graded as good, but requires some improvements and correction before publishing.
•	The professional English language is used throughout the manuscript.
•	The topic of the study is interesting, it fits into trends in science as well as in the manufacturing practice. Submitted paper is compliant with the aims and scope of the scientific journal PeerJ.
•	The title cover the main aspects in this paper, it reflect the aim and scientific purpose of conducted experiment.
•	Presented abstract doesn’t explain the significance of the paper. This part of the manuscript seem to be descriptive.
o	Please rewrite it clearly stating the facts; focus more on how your research has contributed to knowledge gaps; describe research limitations for future research and restate your major findings.
o	Instead of indicatively stating the obtained results, it would be better to explain the reasons for choosing the process parameters and substantiate them with the obtained results. Eg., for samples A, the application of the following production conditions is recommended in order to obtain more desirable quality parameters, which is confirmed by the performed sensory analysis.
•	Introduction provides a good, generalized background of the topic giving the reader an appreciation of the wide range of applications for this technology.
•	The methods used in this paper are appropriate to the aim of the study. Methods are clear and replicable.
•	Results are presented in an appropriate format (clear and legible), well labelled and described. Raw data were supplied as supplementary file.
•	The literature cited in this paper is relevant to the study. The paper use appropriate references in the correct style to promote understanding of the content.
Experimental design: The methods used in this manuscript are standard and appropriate to the aim of the study. Methods are clear and replicable. In this research, no measurement of parameters such as foam stability or bubble size and distribution, microbiological analysis of beer, etc. was performed. These parameters could greatly contribute to the quality of research.
The disadvantage of this paper is that it does not clearly highlight the scientific contribution that the results will bring. It could be said that the research question is not well defined (in terms of defining process conditions of beer production) /Line106-108.
Validity of the findings: Conclusions presented in this paper correlate to the results found. Given the scope of the results presented, it is necessary to improve conclusion section. Please rewrite it and focus more on how your research has contributed to knowledge gaps; add scientific and practical significance of the selected method.

•	Uniform the numbering of the titles and subheadings of the paragraphs.
Line 34: Please check the expression “thousand corn weight”!
Line 40, 47: Correct ABV to %ABV.

M&M
•	In general, the font and the way of writing mathematical equations should be standardized. Some mathematical equation also lack explanations of variables and/or units of measurement.
o	Equations 1, 2, 7, 8 gives the full names of the variables, in Eq. 3, 4 and 5 is a mix, and in Eq. 6 and 9 only the short ones. Furthermore, in Equation 10, in addition to the full name of the variable, its abbreviation is also given. In some equations (eg Eq. 11) the unit of measurement is given next to the variable, while in others it is not. Align it.
•	In some places in the text the term “rice malt” is used while in others it is just “malt”, this needs to be standardized.
•	Where possible, add the number of iterations/determinations of the analysis performed.
Line 171: The title says a thousand grains of corn. Is that a mistake? Should “corn” be replaced by “grain of rice”?
Line 182: It is necessary to explain the variable "Dormancy" in the mathematical equation shown.
Line 191: What is the variable called X. State its units of measurement.
Line 192-195: Please state the units of measurement for the following variables: Wi, Wf, MC, D. There is missing explanation of the variable D.
Line 204: Delete the colon in the paragraph title.
Line 210: Explain the S.G variable. If the authors meant the variable Specific Gravity Degrees, then it should be clarified.
Line 211: Why the title of the paragraph begins with the number 2.4.1.2., when the previous paragraph is numbered 2.5.1.1?
Line 213: Should be written Lintner Degrees per Kilogram (°Lintner/kg).
Line 224-225: If it is a Lintner degree please correct “°L” to “°Lintner”. Lintner is an index that measures a malt’s diastatic power (DP), commonly written as “°Lintner”. To avoid possible misinterpretation of the results (degrees Lovibond or Lintner), it is recommended to write the DP unit of measurement as °Lintner. Please make mentioned correction in the Eq. 5.
Line 269: Add an explanation of the variables TSN and TN.
Line 319: What about measurements performed in triplicates (Line175)?

R&D
Line 538 and 651: Change ABV to %ABV.
Table 3, 4: To avoid possible misinterpretation of the results (degrees Lovibond or Lintner), it is recommended to write the Lintner degree as “°Lintner”. Please apply this change to all Discussion section.
Additional comments: No additional comments