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: PHYLOGENOMIC ANALYSES REVEAL A GONDWANAN ORIGIN AND REPEATED OUT OF INDIA COLONIZATIONS INTO ASIA BY TARANTULAS (ARANEAE: THERAPHOSIDAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is an important contribution to the understanding of the systematics and biogeography of Theraphosidae spiders. Although not many new sequences are generated, the authors compile a meaningful number of sequences from online repositories and reanalyze them under the lens of new question, such as are theraphosids of Gondwanan origin? What is the origin of Asian Theraphosidae?

All sections of the manuscript (intro, methods, results and discussion) are well written, but several minor issues should be addressed before publication, a list of which can be found below:

Line  109: correct  “dating to dating”

Lines 149-150: “Given that the theraphosid tree from Foley et al. (2019) received 100% bootstrap support at almost all nodes, we used this as the reference tree to compare with each gene tree.” Phylogenomic and transcriptomic analysis usually get 100% bootstrap support due to the massive amount of data that always support a given topology. One way to assess if the highly supported branches are found across the genes is to run a concordance factor analysis (performed by IQTREE). It is quite simple, and you can use your data in the format as is: http://www.robertlanfear.com/blog/files/concordance_factors.html

Line 169: “Rosamygale grauvogely” should be in italics.

Line 195: “(A) Indian 195 Subcontinent; (B) Asia; (C) East of the Wallace Line; (D) Americas; and (E) Africa”
These biogeographic areas were delimited based on which criteria? Did you use a reference from literature? There is plenty of references showing the importance of delimiting reasonable biogeographic areas, e.g. https://www.researchgate.net/publication/322820908_Biogeographical_units_matter
The biogeographic areas indicated as ABCDE are used only in the Supplementary Table 1 and supplementary figures. Why not use it in the text and in Figure 3? It will surely be helpful and will make the results more intelligible. While at it, the Supplemental Table 1 could also include a column with a more precise location for the species (at least at country level), not only the geographic range.

Line 201: “via model testing (Table 1) BioGeoBEARS”
There is something missing here, it should probably be “in BioGeoBEARS”

Lines 205-206: Although the “j” parameter have been criticized recently, it has also received support and further detailing in its use: https://onlinelibrary.wiley.com/doi/abs/10.1111/jbi.13496; https://academic.oup.com/sysbio/article/69/1/61/5490843

Line 213: Ages of theraphosid clades
The first paragraph of this section contains a mix of information. The first sentence is about the phylogenetic position of Rhianodes atratus (which is mostly expected), and the rest of the paragraph is about incongruences between the biogeographic analyses, but no dates are mentioned at all. I suggest this paragraph to be moved somewhere else in the text.

Line 224: “is recovered between 120Ma and 115.5Ma in the Americas (Figure 3).”
Figure 3 doesn’t show the variance bar, that is Figure 2.

Line 227: “The Indian Plate is first represented between 108Ma and 103.5Ma (node 25).” The Indian Plate is not represented in your dating analysis, the age of the node is what is being recovered in agreement with the position of the Indian Plate. Please rephrase.

Line 227: Here the text gets a bit confusing because the node numbers are shown in Figure 2, while the image with a better resolution of species names and with the ancestral areas reconstructed in nodes are in Figure 3. So, when you say “The first such node (26) emerges between 99Ma and 95Ma” I can see in figure 2 that node 26 in over that date range, but I don’t know which ancestral area it occupies, so I move to figure 3, but there’s no node 26 in figure 3. So, I have to go back to figure 2, lean in to try to read the species names, go back to figure 3, locate the clade and finally visualize what the biogeography of the group is. This kind of back and forth of figures happens during the whole results and discussion. Maybe this could be simplified by adding the node numbers in Figure 3.

Lines 227-235 (and in several parts of the text): “between 108Ma and 103.5Ma; between 99Ma and 95Ma; between 57.4Ma and 55Ma; between 47.5Ma and 45.5Ma”.
All this can be written in a more succinct way with a range, such as 108¬–103.5 Myr, 99–95 Myr, 57.4¬–55 Myr …

Line 248: “The event matrices of all analyses for this node are given in Table 2.” This should be in the methods section.

Line 261, 262: Please, add node numbers to Figure 3, it will make the life of readers much easier. Another option is, when citing Figure 3, not referencing node numbers at all, because we can already guide ourselves with species and clade names.

Line 311: “which split off quite close to the root of the tree”
Catumiri does not split off close to the root of the tree (it does only in figure 3, which is a crop of the larger phylogeny, which includes the actual root of the tree).

Figure 1: This figure doesn’t have much information (like distribution points) so there’s no reason for it to be this big. I suggest placing it, reduced in size, together with Figure 3, this way it will easier for the reader to associate the colors of the biogeographic regions with the colors on the tree.

Figure 2: the years in the time scale are negative and should be corrected to non-negative values.

Figure 3: Maybe change the star to another symbol to represent independent colonization of Asia; you already use starts to represent calibration nodes in Figure 2 and it gets confusing as readers reach figure 3 with a preconception of the meaning of a star in a dated phylogeny.

All supplementary tables should have legends!

Supplementary table 4 contains information that are too important to be buried into supplementary materials. It should be reformatted and included in the main paper (at least the theraphosid part of it).


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: CONTENTS AND YIELDS OF COPPER, IRON, MANGANESE AND ZINC WOULD BE AFFECTED BY LUCERNE AGE AND CUT
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: in the experimental design, authors defined the age, cut of the lucerne, but these definitions should be detailed introduction as the results.
Validity of the findings: no comment
Additional comments: This manuscript presented the changing microelements of lucerne with stand ages, cuts and organs. Authors verified the characteristic of the changing factors. These results have certain significance to understand the growth law of lucerne. But there were also some questions as follows. Thus, the manuscript can be accept after correct these questions.
1，in the experimental design, authors defined the age, cut of the lucerne, but these definitions should be detailed introduction as the results.
2，Please correct the number of decimal places for uniform data retention in same line of tables.
3，The headings were too detailed in result, please combine and redescribe them.


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: CONTENTS AND YIELDS OF COPPER, IRON, MANGANESE AND ZINC WOULD BE AFFECTED BY LUCERNE AGE AND CUT
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comments.
Experimental design: no comments.
Validity of the findings: no comments.
Additional comments: Line33：”cutting period” must be given the specific growth stage, maybe”early flowering stage”?
Line 37: or should be changed as “and”.
Line 27 and 43: Unified microelements or trace element.
Line 82: “plateau” changed as “Plateau”.
Line 83:“480mm to 660mm”changed as “480~660 mm”.
Line 91: the cutting period? When or which stage?
Line 102-103: “ml” changed as “mL”.
Line 109: Three-way analysis? The correct description maybe Multivariate analysis of variance analysis?


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: CONTENTS AND YIELDS OF COPPER, IRON, MANGANESE AND ZINC WOULD BE AFFECTED BY LUCERNE AGE AND CUT
Review round: 1
Reviewer: 3

Output:
Basic reporting: -	The English language could be improved to ensure that an international reader can clearly understand your text. Some examples where the language could be improved include but not limit to:
	[line 29-30] It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 30-31] What do you mean by “stand age”? It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 33-34] It could be changed to “the contents and yields of Cu, Fe, Mn and Zn in lucerne”.
	[line 35-36] It’s a little confusing when you use “respectively” here. It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 36-37] The same “respectively” issue, it would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 46] “basal”  “basic” Is that what you mean to say?
	[line 48] It’s a little bit confusing about the use of “and” and “but” in this sentence. It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 58] “Microelement as macroelement” could be changed to “Similar to microelement, microelement…”
	[line 60] delete the “and” at the end of this line
	[line 61] It’s a little bit confusing about the use of “As well” in this sentence. It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 66] The “ha” in the phrase “32 million ha” could be replaced by its full name “hectare”, because this is the first time to use it. Then you can choose to use abbreviation later.
	[line 94] It’s a little bit confusing about the use of “secondly” in this sentence. It would be helpful to rephrase this sentence to allow readers to better understand it.
	[line 99] “digestive tube”  “digestion tube”
	[line 100] It would be helpful to mention the concentration of nitric acid you used.
	[line 101] It would be helpful to describe more details about your chemical analysis. For example, you could provide more details on your digestion method and parameters you used (e.g., temperature, power, use gradient program or not, etc).
	[line 103] “solution of beaker”  “sample solution”
	[line 104] It’s a little bit confusing about the use of “at a constant volume” in this sentence. Do you mean you measured and recorded the volume?
	[line 104] “ice” needs to be replaced by its full name. And please detail your analytical method and parameters when you use this instrument. It would be very helpful for the readers to reproduce your work.
	[line 211] “most positive correlations were found” seems a little bit vague. It would be better to restate your results and conclusions more clearly.
Experimental design: In this paper, the research question was relatively well-defined and meaningful. Both experimental and statistical investigation have been performed. However, it would be more helpful to add more description and information to help readers to better understand on the experiments and results.
Validity of the findings: The results are valid and useful.
Additional comments: It would be helpful to update your references and cite more recent research to replace relatively old ones.


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: CONTENTS AND YIELDS OF COPPER, IRON, MANGANESE AND ZINC WOULD BE AFFECTED BY LUCERNE AGE AND CUT
Review round: 1
Reviewer: 4

Output:
Basic reporting: Please see my comments in "General comments for the author"
Experimental design: Please see my comments in "General comments for the author"
Validity of the findings: Please see my comments in "General comments for the author"
Additional comments: The article presents a study of contents and yields of copper, iron, manganese, and zinc
in lucerne of different ages and cuts, as well as the differences among organs. Statistical analyses were also employed to confirm the alteration. Overall, the language is fair, and the manuscript is generally well-organized.

Major scientific comments:
1.	In the “Introduction” section, the motivation of this study is unclear. It seems that the authors did this study because “the elements play important roles, so we want to know the alterations”. The authors had a small paragraph (line 58-64) discussing the relationship between microelements and plant growth and the health of livestock. This might be a meaningful reason that drives this study. However, the authors just huddled thru with limited examples. It is suggested to include more details/examples in this paragraph to interest readers. The authors should introduce more on the relationship of microelements and plant growth/animal health.
Similarly, in the “Conclusions” section, authors should give some perspective on how the current study benefits, but not just give a summary of the results.
2.	In the Methods section, line 98-106, the authors employed a digestion&spectrometer-based method to analyze the chemicals. Is this method previously reported? If so, please cite it. Otherwise, authors need to do some method validations, e.g. linear range, LOD/LOQ, accuracy/precision. It is important to know these parameters since some of the results differed from the literature, which was discussed in the “Discussion” section. Note if the method is not accurate for the analytes, it will also lead to differences.
Line 105-106, the authors claimed, “Lastly, the contents were calculated by a formula.” Is this “formula” well-known? Or is it created by the standard curve made in house? Please cite or show the formula if it is known. Otherwise, if it is built by a standard curve, authors need to indicate the standard chemicals they used.
3.	Significant correlations among the biomass, contents and yields of Cu, Fe, Mn and Zn were shown in Table 7. It is suggested to move this paragraph to the “Discussions” section. Moreover, there is no single reference in this paragraph when the authors discussed the correlations among elements. Instead, the authors used ambiguous sentences “ The Fe content increased with the Mn and Zn contents, and it positively contributed to the accumulations on yields of Fe, Mn and Zn.” I assume there are plenty of studies regarding relationships of element up-taking in the literature. The authors should find and cite them instead of just guessing.

Minor comments

4.	In Table 4-6, lowercase letters “a” and “b” were shown. Their meanings were not clearly annotated.  I assume they are referred as “ The significant difference was determined by one-way ANOVA, and it was represented as the different lowercase letters (P<0.05).” Additionally, the authors need to clarify what the difference is between two lowercase letters. Probably, one for p<0.01 while another for p<0.05?


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: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 1
Reviewer: 1

Output:
Basic reporting: The Manuscript is clear and easy to understand.
Experimental design: Experimental design was good but I feel there was scope of improvement.
Validity of the findings: Conclusions are well stated and according to the study's finding.
Additional comments: Dear Authors,
First of all, I would like to congratulate you for the fantastic work you have done in this study. The manuscript by Diaz-Velez et al. discussed the serological surveillance, based on the measurement of the presence specific antibodies (the test kit used in this study was approved by FDA) against SARS-CoV-2 in a given population in Lambayeque region of Peru. Population-based serosurveys are a powerful source of information to monitor the progression of the epidemic. The authors have examined the presence of IgG/IgM antibodies from 2010 people. The results and conclusions of the papers are interesting and important. I also find the study to be relevant and well written. I liked reading this paper, and the text description of the results was convincing. However, I found some concerns that need to be addressed before considering this manuscript for publication.
1.	Why were the individuals below the age of 9 not included in the study? This study may not be the representative of the population.
2.	The percentage of participants that had previous of SARS-CoV-2 PCR testing was not reported in this study therefore it is not clear whether or not there was a bias for probable cases.
3.	What measures had been taken to differentiate between a true and false positive?
4.	I am wondering, were the authors considered repeated measurements of antibodies at different time points for the seroprevalence estimation? Antibody titers may decline with time and this would give falsely low prevalence estimates.
5.	It could have been better if the authors would consider the gender, body mass index and smoking habit as a trait for figure 2C.
6.	Did the authors estimate the infection-to-fatality ratio of COVID-19 in the given population?


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: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 1
Reviewer: 2

Output:
Basic reporting: -	Please cite a reference for lines 48-50.
-	Lines 60-64 need to be divided into multiple sentences. The sentence construction is not ideal.
-	The introduction section needs more information. This includes: What is the prevalence of false positives in the serological COVID tests? What is the population division by age, gender, preexisting conditions in the Lambayeque region? What are the possible causes for almost 88.7% of Peru’s cases being asymptomatic?
-	Line 241 should be “this finding” and not “his finding”. Please correct.
Experimental design: This was an exhaustive study in a set-up which is ever changing and therefore difficult to conclude. The authors have done a commendable job considering the limitations, however there are some major aspects which have not been listed in the experimental design:

-	For the selected population, what percentage had preexisting conditions asper the collected questionnaire? Was this confirmed with medical records of the patients?
-	Both IgG and IgM positive results were considered. Explain how the results between the 2 antibodies could vary and how they could lead to different conclusions.
-	For individuals who did not practice strict isolation, was the use of masks taken into consideration? How did that affect the results?
-	Were any of the seropositive individuals tested with qPCR too? Would that have helped in having greater faith in the results?
-	Was an orthogonal testing algorithm as recommended by the CDC employed for this study?
Validity of the findings: -	The findings of the study though interesting, are based on experimental design which has scientific omissions and lack of rigor. Hence it is difficult to look at the findings without taking these errors into consideration.
-	Statistical analysis is thorough and reliable.
Additional comments: Listed in the sections above.


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

Input:
Title: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article describes the rationale and the methodology of the study in sufficient detail. The message that the article wants to convey can be much easier to elucidate if grammatical errors are addressed. For instance, lines 59-64, 134-142 needs to be rephrased. Authors should also address the typographical errors in the text.
Experimental design: The experimental design is well-thought-of. It will be nice if the authors could show some kind of validation for some of the positive tested samples using conventional qPCR. This will help better in assessing the false-positive or false-negative rates.
Validity of the findings: The authors have not commented on the false positive and false negative rates for their analysis. It has been reported that some of these tests do have a kind of false positivity or negativity. Did the authors observe this in their analysis?
Additional comments: Authors have very nicely demonstrated the SARS-CoV-2 seroprevalence study in Lambayeque, Peru in the manuscript and have reported their analysis following proper statistical tests. The manuscript is suitable for publication, provided the above comments are addressed.


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

Input:
Title: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 2
Reviewer: 1

Output:
Basic reporting: The Manuscript has improved considerable after the authors considered the reviewer's suggestion.
Experimental design: Authors were acknowledged the limitations of the study in the revised version.
Validity of the findings: Conclusions are well stated and according to the study's finding
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: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 2
Reviewer: 2

Output:
Basic reporting: The language has been corrected where necessary and typographical errors have been addressed.
Background section is robust and authors have duly added information that was previously asked.
Experimental design: Robust experimental design. The lack of qPCR testing has been considered as a limitation.
Validity of the findings: Data is statistically sound and adequate controls have been used.
Additional comments: The authors have a done a sound job with addressing previous 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: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 2
Reviewer: 3

Output:
Basic reporting: The language can be better
Experimental design: It needs some more rigorous analysis as I will highlight later
Validity of the findings: This is a well done paper but could be made better.
Additional comments: The authors do a good job of rebutting the reviewers. However, I would like to ask some more analysis from the authors
1) a)"The high seroprevalence reported in our research could be due to the identification social isolation measures and 27.1% have visited a market 14 days prior to the investigation. "
b) "This is because 25.9% reported that they have not maintained the mandatory that could explain the differences found is the method used to detect antibodies against SARS-CoV-2, which has lower sensitivity in contrast to ELISA or chemiluminescence tests (Lisboa Bastos et al. 2020)."
Can the authors support both the above statements with more rigorous analysis as opposed to just percentages? This analysis can then extended to other such factors which can explain the high seroprevalance.
2) How much did the past medical history of the sample size influence SARS-COV2 positive test rate? Is it possible that the general population was less healthier before SARS-COV2, which caused more people to fall sick, and thus the high seroprevalance?

Minor comment
1) Language should be brushed up


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: SARS-COV-2 SEROPREVALENCE STUDY IN LAMBAYEQUE, PERU. JUNE–JULY 2020
Review round: 3
Reviewer: 1

Output:
Basic reporting: No issues
Experimental design: Good
Validity of the findings: Good
Additional comments: The authors did not quite respond to the issue I raised especially point 1. However, the manuscript is good and can be accepted.


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: COMPARATIVE ANALYSIS OF MAPK AND MKK GENE FAMILIES REVEALS DIFFERENTIAL EVOLUTIONARY PATTERNS IN BRACHYPODIUM DISTACHYON INBRED LINES
Review round: 1
Reviewer: 1

Output:
Basic reporting: Comment 1: Line no. 283-286 says, “To explore the expression variation of the MPK and MKK gene family in Bd21, BdTR8i and Bd30-1 that belongs separately to three different genetic groups, the expression profiles for five different tissues (root, stem, leaf blade, leaf sheath and spikelet) of MPK and MKK genes were preformed”. The authors have performed the expression analysis of all the MKKs and MPKs in the given Brachypodium inbreed lines for five different plant tissues. The MAPK genes are involved in various abiotic stress tolerance responses and developmental processes, which are justified by Goyal et al., 2018 in his paper “Analysis of MAPK and MAPKK gene families in wheat and related Triticeae species”. They have written, “Mitogen-activated protein kinases (MAPKs) are key signaling enzymes involved in the regulation of various aspects of biology in eukaryotic organisms, including cell division, development, metabolism and stress responses”. The authors should consider providing some abiotic stress tolerance data concerning their identified MKK and MPK genes based on the pivotal importance of the MAPK signaling genes in abiotic stress tolerance mechanisms.
Experimental design: Comment 1: Line no. 90-93 says, “To identify these genes in the 53 diverse B. distachyon inbred lines, BLASTP (Altschul et al. 1997) searches were performed by orthologous protein sequences using BdMAPKs and BdMKKs as the query search in BrachyPan (https://brachypan.jgi.doe.gov/) (Gordon et al. 2017)”. The authors have stated the use of orthologous genes for blast-p analysis. A pint-sized explanation can be provided here in this section for doing the same.
Comment 2: Line no. 111-113 says, “Phylogenetic trees based on the alignment of all MAPKs or MKKs were conducted using the maximum likelihood (ML) method with the Jones-Taylor-Thornton (JTT) model, 2000 bootstrap values and partial deletion by the MEGA 6.0 software”. Why consider using 2000 bootstrap replicate values rather than a 1000? An explanation can be provided for the same.
Comment 3: The authors have used the Maximum Likelihood (ML) method to construct the phylogenetic trees and have represented the trees across figures 1 and 2. The authors can improvise on the representation of the trees. The huge number of genes for both the MKKs and MPKs have created a pile of gene ids in a confined space, which is almost impossible to figure out. The authors can also consider the use of Neighbor Joining (NJ) algorithm to represent their phylogenetic trees and provide them in supplemental files.
Comment 4: Line no. 261-263 says, “Gene duplication was considered as the primary force of the species evolution, which resulted in the gene to form the gene families. In addition, some tandem duplication was observed in monocots MKK10 paralogues, such as in B. distachyon (Jiang and Chu 2018)”. The authors should provide a little more clarity in the above-quoted sentence.
Validity of the findings: No comment
Additional comments: Dear authors, firstly I would like to appreciate the hard work that has been put together to compose this manuscript. To better understand the evolution and function of MAPKs and MKKs in B. distachyon inbred lines, the authors conducted their systematical molecular evolutionary analysis among the 799 MAPKs and 618 MKKs, which were identified from 53 B. distachyon inbred lines, respectively. Phylogenetic analyses and intron-exon patterns were analyzed followed by the expression analysis of all the MKKs and MPKs. These findings can provide new insights into the functional evolution of genes in closely inbred lines. Nevertheless as a reviewer of the scientific community, These are some of the aspects of the manuscript, which can be touched upon and improvised.


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: COMPARATIVE ANALYSIS OF MAPK AND MKK GENE FAMILIES REVEALS DIFFERENTIAL EVOLUTIONARY PATTERNS IN BRACHYPODIUM DISTACHYON INBRED LINES
Review round: 1
Reviewer: 2

Output:
Basic reporting: No Comment
Experimental design: No Comment
Validity of the findings: No Comment
Additional comments: The manuscript entitled "Comparative analysis of MAPK and MKK gene families reveals
differential evolutionary patterns in Brachypodium distachyon
inbred lines" submit by Jiang et al. is a robust analysis of MAPK and MKK gene families in 53 inbred lines of Brachypodium distachyon. The analysis is good and presentation of the entire manuscript is also impressive. The authors have identified a total of 799 MAPKs and 618 MKKs in 53 inbred lines of B. distachyan, respectively. Theses MAPKs and MKKs are divided into four groups as reported earlier in Arabidopsis and other plants. They also analysed the expression of these genes in different tissues. The highlight of the study is  revealing the role UTRs on structure and evolution of MPK21-2 genes and differential evolution of MKK10 paralogues with ancient tandem duplication. I overall liked the manuscript, however I have following very minor concerns that authors may choose to attend:
1. I would prefer to highlight the specific numbers of MAPKs(14/15/16) and MKKs(12) members in each lines of B. distachyon in the abstract.
2. Since authors have performed MSA of 53 different inbred lines of B. distachyon, I was looking for a conserved domain specific to B. distachyon  members compared to other plant species. If possible, this information may be added in Figure 5.
3. Authors may provide a bit more details about the age of plants used from which tissues were harvested for expression analysis in the legend of Figure 7.


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: COMPARATIVE ANALYSIS OF MAPK AND MKK GENE FAMILIES REVEALS DIFFERENTIAL EVOLUTIONARY PATTERNS IN BRACHYPODIUM DISTACHYON INBRED LINES
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The Authors have appropriately modified the revised version of the manuscript. I have no further question for this article.


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

Input:
Title: BEHAVIORAL RESPONSE OF PANONYCHUS CITRI (MCGREGOR) (ACARI: TETRANYCHIDAE) TO SYNTHETIC CHEMICALS AND OILS
Review round: 1
Reviewer: 1

Output:
Basic reporting: I have not reviewed the complete manuscript due to very poor English. At several instances I was unable to understand whole sentence or even whole paragraphs (see result section). Please see my comments in the attached file
Experimental design: Although, the experimental design is valid and it need many additional information that should be added in the manuscript to make it clear and understandable for the readers. Please see my comments in the attached file
Validity of the findings: Please see my comments in the attached file
Additional comments: Please see my comments in the attached file


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

Input:
Title: BEHAVIORAL RESPONSE OF PANONYCHUS CITRI (MCGREGOR) (ACARI: TETRANYCHIDAE) TO SYNTHETIC CHEMICALS AND OILS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Throughout the manuscript there are some inconsistencies with respect to the figures' calls in the text. Please review them.
Experimental design: No comment
Validity of the findings: No comment.
Additional comments: The manuscript is very detailed, however, it is necessary to make some adjustments that were verified throughout the text to improve comprehension.

Line 111-113: In the methodology mentions that the CL'30 and CL'50 were used, where did these values come from? A previous experiment was carried out? Or are they data from the literature? Please rewrite to clarify this issue.

What type of vegetable oil and why was it used? Justify to improve understanding.

Line 149-151: The information is not clear or is not consistent with the result shown in figure 2B, where there was no significant difference between the CL30 and CL50 for both SYP-9625 and EnSpray 99.

Line 152-153: This information does not match as shown in the figure. In the caption of the figure lowercase letters = difference between CL 30 and 50. In figure 2C The CL's of EnSpray 99 do not differ. Please review! And change Fig. 1C to Fig. 2C (line 153).

Line 155: “Fig. 1D ”does not match what is written.

Line 346: The conclusion of the manuscript item 2 mentions that the preferred leaf surface was adaxial. Probably the largest amount of mites recovered on the right adaxial surface is related to the release point, this may have tended the number of recovered on the adaxial surface.You could justify in your methodology why there was only this point of release. For the ideal would be to either release in alternate points or an intermediate point between the two areas: right and left.

Figure 2 legend: I suggest you insert what blue and white means in the captions of the figures.

Figure 3 – 9 legend: Please add in the caption what *** and “ns” mean.

Congratulations on the 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: BEHAVIORAL RESPONSE OF PANONYCHUS CITRI (MCGREGOR) (ACARI: TETRANYCHIDAE) TO SYNTHETIC CHEMICALS AND OILS
Review round: 2
Reviewer: 1

Output:
Basic reporting: See my comments on the attached file
Experimental design: See my comments on the attached file
Validity of the findings: See my comments on the attached file
Additional comments: See my comments on the attached file


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

Input:
Title: MULTI-SCHEMA COMPUTATIONAL PREDICTION OF THE COMPREHENSIVE SARS-COV-2 VS. HUMAN INTERACTOME
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript investigates interactions between the proteins of the SARS-CoV-2 virus and human proteins. Two top protein-protein interaction (PPI) predictors are used and further efforts are made to identify the most confident PPIs. In addition, the prediction are enhanced by the identification of the subsequences that medite the interactions. The main goal is to provide additional information for the community that can be useful in understanding and disrupting its host-pathogen mechanisms.

The manuscript is clearly written, provides appropriate references, includes relevant
figures and makes its data available.
Experimental design: The research described is very well within the scope of the journal. The research is clearly described and important for the problem investigated. The methods are new and interesting, well presented and reproducible.
Validity of the findings: Findings are important, valid and potentially very useful for a large community. To the extent that they overlap, they agree with existing knowledge about the virus.

The overall evaluation of the manuscript is highly positive. It provides an interesting,  useful and timely contribution to a subject of high importance.
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: MULTI-SCHEMA COMPUTATIONAL PREDICTION OF THE COMPREHENSIVE SARS-COV-2 VS. HUMAN INTERACTOME
Review round: 1
Reviewer: 2

Output:
Basic reporting: On page 5, the Kneedle algorithm should be referenced: DOI: 10.1109/ICDCSW.2011.20

In Figure 2, I imagine that the dotted line represents the knee? This should be annotated.

Line 350: "lifecylce" should read: "life cycle"

The color scale of Figure 6 could be adjusted such that differences between the values are more visible. It seems like little to no values are above 3, which could be the maximum of the color scale.

The unit/nature of the values on the legend of Figure 6 and 8 are not reported.

Line 446: there should be a comma before the word "which".
Experimental design: While I am sure that there is a reason for this, why were only 14 proteins of SARS-CoV-2 used for the protein-protein interaction predictions. This should be justified in the manuscript.

This is not a request per se, but I am curious whether the Kneedle algorithm strategy could have been replaced with a null model assessing the statistical significance of a given confidence score. For instance, one could shuffle the protein sequences provided as input and compare the confidence scores of the predicted interactions of those shuffled sequences to the interaction predictions obtained using real sequences. Comparing the number of interactions passing a given confidence score threshold could provide an estimate of the false discoveries. While what I am proposing is a rudimentary approach and that a more sophisticated null model could certainly be designed, I was wondering whether such a general approach would make sense? I would be curious to know what the authors think about this. This could potentially be a discussion point if the authors believe that it is relevant.
Validity of the findings: I understand that there is no overlap with the predictions performed in this manuscript and the Gordon et al., 2020 study of the SARS-CoV-2 and human interactome. However, I was curious to see if there is any overlap in terms of the GO terms identified in this study and the Gordon et al. study. Indeed, many of the interactions reported by Gordon et al. may not be direct interactions due to the nature of the experimental protocol performed. One could easily imagine that a predicted interaction in this study may be involving other indirect interactions that were detected by Gordon et al. Hence, while the interactions do not overlap, the GO terms in which the proteins are involved in may overlap, since some of the proteins may be implicated in similar processes or machineries. Reporting such an overlap analysis would be interesting.
Additional comments: This is a very well written manuscript describing a comprehensive study that is of interest for the community performing research on SARS-CoV-2. The methods are sound, based on well-established tools, PIPE and SPRINT, and the work is also of high interest for the field of protein-protein interaction prediction in the context of poorly characterized organisms. The study provides information that could be used in the future for the development of therapeutics against SARS-CoV-2 and even other coronaviruses.


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: THREE NEW LUTICOLA D.G.MANN (BACILLARIOPHYTA) SPECIES FROM RAPA NUI (EASTER ISLAND) FOUND IN TERRESTRIAL DIATOM ASSEMBLAGES DOMINATED BY WIDELY DISTRIBUTED TAXA
Review round: 1
Reviewer: 1

Output:
Basic reporting: see additional notes under general comments.
-- Some editing of sentence structure, grammar, correction of typos will need to be completed. Otherwise, the writing is clear.
-- Sufficient and appropriate references are used. Refs – check for italics of genus and species names. (also spacing, use of periods after years, colon versus comma after journal volume, etc.).
--Manuscript goals were laid out.
Experimental design: The manuscript contains original primary research and contributes to knowledge on diatom taxonomy with respect to the genus Luticola. See notes below for how authors can strengthen their description of the research gap.
-- Authors provide appropriate detail and descriptions for the new species proposed.
-- Image plates are professionally laid out and conform to discipline standards.
-- Though the sample size is limited, the authors explain the origin of the samples and identify the limitations in the discussion.

1)	Methods – Suggestions for additional details needed in the methods section.
a.	Include a general map of the global location of the island and sampling site. Is there an image of the cave?
b.	Sample collection: Where in the cave was the sample collected (floor, wall, etc).  Was the sample analyzed from the soil associated with the plant collection, or was it collected with diatoms in mind (it appears as if it was associated with the plant collection).  Provide additional detail here.
c.	L135 – replace ‘community’ with ‘assemblage’ (community implies multiples trophic levels). Do a check throughout the manuscript.
d.	Provide information relevant to new species descriptions. How many valves/frustules were measured under the LM for morphometric descriptions? How many under the SEM for ultrastructure?
e.	Include the literature used for identifications and for the diatom terminology. Consider using parallel terminology for raphe ends: for example, you call distal raphe ends “distal”, but the “proximal” raphe ends “central”.
Validity of the findings: Results are valid. Data provided, include appropriate sample analysis and image documentation. Diatom repositories identified. Discussion is appropriate and links to research goals and is within the bounds of the results.
Additional comments: General comments
The authors use light and field emission microscope evidence from analysis of a diatom sample from a cave on the isolated island of Rap Nui (Easter Island) to describe and document three new species of Luticola (L. georgzizkae, L. rapanuiensis and L. moaiorum). These new taxa are compared to another Luticola taxon present in the sample, L. ectorii, and other Luticola species. The diatom diversity at this site, reported as low, contained other taxa common to aerophytic habitats. These descriptions and documentation of Luticola add to our understanding of this diatom genus and the diversity in soil-associated diatom floras. The remote nature of the island provides information relevant to diatom biogeography that the authors discuss.

The article will be of interest to readers of the journal Peer-J, especially those interested in diatom taxonomy, Luticola, biogeography. And cave, aerial and limno-terrestrial habitats. I enjoyed this manuscript.

2)	Abstract:
a.	 L29: Add in a little more information about the type of samples/location within the cave (e.g. soil from a plant collection on the floor of the cave? Near the entrance, etc.). This will tell the reader a little more about the type of sample collected in the abstract before article download.

b.	L36 – Include here the information the descriptions are based on (e.g. external and internal morphological features from light and field emission scanning electron microscopic analysis; versus genetic analysis)

3)	Introduction.
a.	The first part of the introduction provides a brief history of the study of diatoms on Rapa Nui. It would strengthen the case to flesh out this introductory paragraph to set the stage for the diversity documented to date, and any habitats that remain to be studied (inland areas??? statues?  caves? Other? - are caves common in the inland or the coast?). Do you expect endemism or common taxa at these sites (why/why not….you hint at some thoughts/evidence here).
i.	Of the freshwater floras examined to date, did the taxa documented contain new species?  For example, L44-45 – How many of the 70 taxa were new to science? common?
ii.	Were the diatoms from the core diverse? Any new to science?
iii.	Are there other inland habitats, such as caves or the stone statues where the diatom flora remains underexplored? Given the remote nature and history of this island, the diatom flora of the sites studied to date, do you expect these habitats to contain other new taxa or mostly common taxa?
iv.	…why this location…why now.. (It’s an interesting habitat and a unique location… …)

b.	L50 – start a new paragraph here. You will need to add a connector set of sentences to go from the introduction of the island to the introduction of Luticola.

c.	Carefully consider the use of the term aerophytic to confirm this is the correct term. There are a number of terms out there that describe different types of aerial to terrestrial environments, for example depending on the moisture source. There is not enough information here to determine if aerophytic is the right term. You associate the term soil in parentheses beside the word and reference that samples were collected from a cave.  Take a short paragraph in the methods or intro to shore up this definition and to clear up how you use the term throughout.

4)	Results.
a.	New species descriptions are consistent and appropriate (watch for typos). See some comments below for strengthening the connection between the descriptions and the image plates.  It is great to have a nice LM size series for each new taxon, and to have multiple SEM images for each taxon!  Thank you for taking the time to do that. Plates are well organized and laid out. Consider running a temporary line horizontally across the middle to line up the central area of each specimen (I think you will find a few will get reordered to create the size diminution series).

b.	Summarize the measurements/features of the Luticola species in a table for future comparison or reference. You could also consider adding in measurements for some of the similar Luticola species. For example, like the tables in Pavlov et al. (2009) on Luticola grupcei, or Levko, Metzeltin, Pavlov (2013)

c.	Given the unique location and the expanding studies of aerophyllic/soil-associated habitats, it would absolutely be fabulous to have an additional plate with a representative image of the other taxa.

5)	Results: Figures/Captions.
a.	Overall, I would add a little more detail to the caption to identify features included in the species description. Add in additional arrows.

b.	Fig.1.: Add in a little more detail. A-N – size diminution series (relevant to all captions). Indicate which is the image of the holotype and lectotype.

You indicate the “H” is a spineless (implied SEM given sequence) – but H is an LM.  Reword or re-order phrasing.
(Z) arrow stated in Fig. caption – none on the image (add in)

c.	Fig. 2. Add in a little detail. For example, what do the external views highlight or show? What about the girdle or internal view? (Q-w) did you mean valve? Or vale?  Arrow only on U and V – either add arrows or reword to be clear.  Identify/include the holotype and lectotype images.

d.	Fig. 3 V – it mentions that “V” shows the stigma (looks to be covered with debris)…reference a different image for this feature. Z – mentions arrow – add in the arrow in the image. Identify in the caption and in the image the holotype and lectotype.

e.	Luticola rapanuiensis. L197 did you mean “within”?
I’m not sure undulate is the right word to capture the raphe shape…or maybe broadly undulate or…? (or maybe it is…maybe revisit).  Where you describe the irregular shallow grooves by the proximal raphe ends, do you mean to reference these as ghost areolae? You label them as such in the figure caption, but not necessarily in the description.  Be clear and consistent. You may also want to indicate in the caption that the valve in image V is tilted to skew the view of the valve face/ highlight its topography.

6)	Discussion -

a)	The discussion points seem relevant and provide appropriate comparisons with similar taxa.

b)	The authors may consider making a point on how the source of the sample (e.g. a botanical sample) may provide other unique opportunities to study diatoms from unique or understudied environments. Any plans to continue the study on the diatom flora of Rapa Nui?

7)	Minor/other:
a)	There are lots of inconsistencies around the italics of the parentheses of the in-text citations (some are italicized, others are not, or one is and one is not). Do a careful edit of these.

b)	L44 missing a word after microscopy-based? (or sentence needs an edit). -- Overall, there are numerous typos or minor grammar corrections needed throughout the manuscript. For example., L393 “not exits on” (“do not exist at”); rhomic-elliptic (did you mean rhombic-elliptic); “Levkow et. al., 2013, refs” – should be Levkov; “Rap” Nui versus “Rapa Nui”.

c)	You flip between the use of Rapa Nui and Easter Island. Consider if you want to present the location consistently one way or the other throughout or if you want to use both.  For example, in the discussion on biogeography you use Easter Island, but then for the conclusion, flip back to Rapa Nui. (I might lean toward using the original name..like you do in the title and abstract)


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: THREE NEW LUTICOLA D.G.MANN (BACILLARIOPHYTA) SPECIES FROM RAPA NUI (EASTER ISLAND) FOUND IN TERRESTRIAL DIATOM ASSEMBLAGES DOMINATED BY WIDELY DISTRIBUTED TAXA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Dear Editor,
The submitted manuscript reports diatom findings from a single sample taken from cave on the isolated island Rapa Nui. Among other aerophilic genera, four Luticola species were found in the sample and three out of them are described in the manuscript as new to science.
Experimental design: A single samples was examined for the diatom species composition under the LM and SEM microscopy which is basic and common treatment for the diatom research. Presented species are well documented and described in the text. It is clear, that authors have experience in the taxonomic work and are familiar with the genus Luticola. The important comparison with other similar species is included.
Validity of the findings: The genus Luticola has been recently very popular diatom genus as model organism to study the species distribution and its biogeography. Many new species have been described all over the world during last decides and we can see a high number of endemic species presented only in specific areas. Specifically, in this study, the geographical isolation is so high that it is not surprising that species new to science have been found there.  It is very species rich genus accommodating around 240 species these days. The genus has received many attention after its generic establishment, recently mainly withing the new monograph written by Levkov et all and work done at south-Asia and Antarctica. Many times we can hear that those regions are the diversity  hot spots, however the question is, if that is not influenced just by under-sampling of other localities worldwide. So from that point of view I see very important and I welcome very much any record from other parts of the world presenting the species composition and distribution of the Luticola species.
Additional comments: It is well written manuscript but what would be excellent is if authors treating (not only) this genus would consider including also molecular data of the species as it is the key to uncover the real diversity of the genus Luticola.


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: THREE NEW LUTICOLA D.G.MANN (BACILLARIOPHYTA) SPECIES FROM RAPA NUI (EASTER ISLAND) FOUND IN TERRESTRIAL DIATOM ASSEMBLAGES DOMINATED BY WIDELY DISTRIBUTED TAXA
Review round: 1
Reviewer: 3

Output:
Basic reporting: For the Introduction, I recommend checking the articles “Bustos S., Morales M. R. & Maidana N. I. 2017. Diversidad del género Luticola (Bacillariophyceae) en sedimentos holocenicos de la Puna Jujeña, Argentina. Boletín de la Sociedad Argentina de Botánica 52: 13–25.”, “Straube A., Tremarin P. I. & Ludwig T. A. V. 2017. Species of Luticola D.G. Mann (Bacillariophyceae) in the Atlantic Forest rivers from southern Brazil. Diatom Research 32: 417–437.”, and “DA SILVA-LEHMKUHL A. M., Ludwig T. A. V., Tremarin P. I. & Bicudo D. C. 2019. On Luticola Mann (Bacillariophyceae) in Southeastern Brazil: Taxonomy, ecology and description of two new species. Phytotaxa 402: 165–186.” in which describe new species of Luticola in the Andes and South America rainforest.

I considered that the background about vascular plants is a little bit extensive, maybe you can shorten this.

If you want you can add a map to highlight the isolation of the island.

I think when you talk about a structure is necessary to add references to the figures where the structure can be observed.

Do you have another picture of the internal view of Luticola rapanuiensis? I am not sure about the stigma internal opening, I saw a mark around the opening, maybe is a scar of the stigma structure.

About the stigma internal opening in Luticola georgzizkae and Luticola moaiorum I think that in both species is a circular and lipped structure without a distinct opening that in the pictures is broken, basing me that in the Luticola moaiorum pictures X and Y the “opening” have different shapes.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: I consider that the article is well structured and presents new relevant information to advance the understanding of the diversity and distribution of the Luticola genus. The species are well defined with strong characters that clearly differentiate them from those present in the literature.
I think the article only needs a few minor corrections, please check the pdf file with my 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 PATIENTS DISCONTINUING OUTPATIENT SERVICES UNDER LONG-TERM CARE INSURANCE AND ITS EFFECT ON FRAILTY DURING COVID-19
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: The impact of COVID-19 varies by region. Please describe the impact of COVID-19 around your facility briefly in Introduction section.

Please describe the outpatient rehabilitation program briefly in Materials & Methods section.

Generally older females use outpatient rehabilitation services more frequently than older males. Please describe why the number of older females was less than that of older males in your study in Discussion section.
Validity of the findings: Although Frailty Screening Index is a screening tool for detecting frailty status, it is not the most suitable assessment tool to detect the frailty status change. Authors should describe it as a limitation.
Additional comments: Thank you for the opportunity to review this article. Our health, psychological status and social activity have been significantly affected by COVID-19 pandemic. The study of the effects of discontinuation of outpatient rehabilitation services for older adults during COVID-19 pandemic is an important subject.

Line 157-159
Authors should describe the following sentence in Materials & Methods section.
“Before the Frailty Screening Index survey was used, participants that had aphasia, dementia, were hospitalized, or could not be interviewed over the telephone were excluded (Figure 1)”.

Line 222-225
The following sentence is not necessary in Conclusions section.
“The purpose of this research was to study community-dwelling older adults needing long-term support who utilize outpatient rehabilitation services to shed light on the characteristics of patients who discontinued their use of outpatient rehabilitation services because of COVID-19 prevention efforts.”


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 PATIENTS DISCONTINUING OUTPATIENT SERVICES UNDER LONG-TERM CARE INSURANCE AND ITS EFFECT ON FRAILTY DURING COVID-19
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is clear and well written. The manuscript introduction, background, structure, tables, and figures meet PeerJ standards. The raw data are provided and easily understandable. Indeed, in my opinion, the article needs only some minor revisions to improve its readability (please, see General comments).
Experimental design: This research is within the aim and scope of the journal and the research question is well defined and contextualized within the current literature. The investigation was conducted with the technical and ethical rigor by the journal. Although some minor revisions are necessary, the methods are described properly (please, see General comments).
Validity of the findings: Notwithstanding the relatively small sample size considering the statistical analysis performed to test the primary aim (i.e., binomial logistic regression analysis), I find the primary aim results of this article robust and the statistical analysis sound considering the observational nature of the study. In this regard, I suggest stating the type of study (e.g., observational, prospective, retrospective, etc.) within the manuscript.
As previously mentioned, the raw data are provided and easily understandable.
Overall, the conclusions are well stated, even though I recommend rewording some sentences (please, see General comments).
Additional comments: Line 34. Please, consider changing “Factor analysis”. Although I understand its meaning and it is grammatically correct, “factor analysis” might create misunderstanding due to the statistical analysis called “factor analysis”.

Line 48. Is it refrain from “physical activities” or “outpatient rehabilitation services”?

Line 49-50. Please, consider rephrasing this sentence “Furthermore, the results suggest that frailty status in the discontinued group might worsen compared to the continuing group, with declines in mental and physical function”. Specifically, due to the observational nature of the study, it is difficult to define what is the cause or the effect; hence, terms like “might worsen” and “declines in” might be inappropriate and could be changed in “is worse” and “lower”. Moreover, I do not understand why the authors report declines in mental and physical function? When rephrasing this sentence, please consider Table 3 results.

Line 59-61. Any reference to support this claim?

Line 78. Please, consider rephrasing this sentence “The effect of discontinuation on frailty was examined” (see comment line 49-50).

Line 78-80. Please, consider changing the position of this sentence “and measures were considered to prevent the worsening of frailty status, such as isolation and disuse syndrome”. In the present form, readers might think that the authors have some results regarding the “measures to prevent…“, whereas those measures are only present in this article’s discussion.

Line 98. Please, consider changing the title “Evaluation of frailty” into something that also includes the other concept within the paragraph (e.g., Assessments).

Line 104. How is the “long-term care level” measured? Please, provide the reference if available.

Line 108-109. What was the elbow position (angle) when the grip strength was measured?

Line 110-112. Any reference regarding the methods used to measure the normal walking speed?

Line 114-116. Isn’t this concept “The participants who continued using outpatient rehabilitation were interviewed in September as well” already explain in the previous sentence?

Line 130-131. “to find the factors for the discontinued group” seems incomplete to me. I would expect to find a verb like associated, affect, or predict. Please, consider the term according to the observational nature of the study.

Line 134-136. Please, consider changing the sentence “In addition, the discontinued group and the continuing group were compared against the Frailty Screening Index in April and again in September” into something like “In addition, the Frailty Screening Index of the discontinued group and the continuing group were compared in April and again in September” to clarify that comparisons were performed only between group and not over time.

Line 137. Why the “presence” of frailty? Were the authors referring to frailty levels or scores?

Line 145-146. Please, consider reporting the overall goodness of fit of the binomial logistic regression (e.g., Cox and Snell R square or Nagelkerke R square).

Line 167. Please, consider changing “participants” into something like “participants of the two groups”.

Line 192. Please, consider changing or deleting “From the above results” to avoid giving the impression that the authors are using the results of this study to support the statement that starts with “we believe …”, which, otherwise, I find appropriate within the discussion.

Line 199. Please, consider changing “suggesting” into something less strong and direct.

Line 210. Please, consider changing “will cause” into something less strong and direct.

Line 227-228. Please, consider rephrasing this sentence “Furthermore, ….” (see comment line 49-50).

Figure 1. Please, consider changing “participants measured”. It could be similar or equal to the title that will replace “Evaluation of frailty” (please, see comment line 98).

Table 1. Please, clarify what the grip strength values represent (e.g., average of the right and left arm). If necessary, also edit the method section and Table 2.

Table 3. I have difficulties understanding what statistical test was used to compute the p values in the last row (i.e., 0.087 and 0.281). If Wilcoxon rank sum tests were used to compare the Frailty Screening Index and not the number of participants in each group, I would recommend adding a row with that index’s descriptive information with their p value on the same row.


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 PATIENTS DISCONTINUING OUTPATIENT SERVICES UNDER LONG-TERM CARE INSURANCE AND ITS EFFECT ON FRAILTY DURING COVID-19
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: The time of "5m walking test" of No. 247184 was 141.58 in the Supplementry_file1(raw_data).
Please check again if you should include the data of No. 247184 in the analysis or not.
Additional comments: I confirmed the revisions of your manuscript.
Thank you for your very careful and extensive 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: CHARACTERISTICS OF PATIENTS DISCONTINUING OUTPATIENT SERVICES UNDER LONG-TERM CARE INSURANCE AND ITS EFFECT ON FRAILTY DURING COVID-19
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Thank you for your willingness to address each recommendation and comment I made. I appreciate the time and effort invested in the edits you made. I am satisfied with the authors' responses.


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: HONEY BEES COMMUNICATE DISTANCE VIA NON-LINEAR WAGGLE DURATION FUNCTIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: -Is line 41 missing the word “bee”?
-Line 40 and 41. There should be a full description of the dance itself and how is interpreted to convey resource coordinates. For example, how the orientation plays a role in location information. I wonder if this can be added as a graphical abstract? (See [1] for an example).
-Line 67-69. This sentence does not make sense.
-Paragraph beginning at 99. This paragraph needs to be improved in clarity. Example. “To settle this ambiguity in the dance-distance relationship, we trained bees to visit feeders located at varying distances greater than 1km. We then determined the duration of the waggle dance components: waggle phase and return phase.” Then add how this data was analyzed to show the non-linear relationship.
-Line 126. Spelled shellac.
-Results Section. It will improve readability if the results are stated to be reported as mean ± std, then simply report the numbers without restating mean±std every time the numbers are given. “We report results as mean±std.” Subsequent results are just “2.20±0.19”.
-Line 401. Please consider the verbiage. “We suspect these later studies to be limited due to the shorter distance of the resource and thus their results are applicable to distances less than 1km.”

[1]  Automatic detection and decoding of honey bee waggle dances
Wario F, Wild B, Rojas R, Landgraf T (2017) Automatic detection and decoding of honey bee waggle dances. PLOS ONE 12(12): e0188626. https://doi.org/10.1371/journal.pone.0188626
Experimental design: -Can the authors add a sentence or two describing how the quality of the resource (feeder) was maintained for the experiment, given that this affects dance performance? This is indicated in the discussion but having it here will quash doubts early on in a reader.
-Could the authors also clarify local feeder conditions (i.e. heavy wind, dim sunlight, competing foragers from native hives, hornets)? Adverse feeder conditions will decrease the bee’s perception of the food source and can affect the return duration.
-I am also curious to know if there were tremble dances from the painted bees as the distance increased. Tremble dances are known to occur in order to inhibit recruitment to a resource.
Validity of the findings: The data analysis appears straightforward and the csv file is readable.
Additional comments: -Citation format seems to randomly change (i.e. line 207).
-Might be a good idea to join the Figure 2 plots. Since the distance is not scaled the same per plots, they look comparable on first sight when they are really not. Perhaps try dashing either von Frisch’s data or Wenner’s data so they can be easily distinguished if joined?

This is a simple yet informative study that clarifies the relationship between waggle dancing and the distance information it conveys. I welcome this study as it clarifies suspicions myself and colleagues had at the time. We suspected the location information to be conveyed in a logarithmic fashion (in line with the flattening theory) but this study suggests a sharper break in distance information. This is very useful information for decoding foraging sites and when interpreting dance information from training experiments.

I would like the authors to seriously reread and revise the verbiage in this paper. For instance, line 90 could be simply written as “It is unclear whether Wenner’s results contradicts the data of von Frisch and Jander, as Wenner’s distance of 1230 m could be too short to detect the waggle duration-distance curvature.”


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: HONEY BEES COMMUNICATE DISTANCE VIA NON-LINEAR WAGGLE DURATION FUNCTIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The article is well-written, well-structured, and flows smoothly from introduction to conclusion.

The only very minor issues I noticed were two sentences that were a little unclear:
Line 344: "When the same group of bees shall be observed dancing for feeders at several distances"- was the meaning that "To ensure that the same group of bees dance for feeders at several distances"?
Lines 365-366: " and little informative with respect to distance"- was the meaning here to say that the return phase duration doesn't convey much information about the distance advertised?

And a few typos:
Line 209: "it was done" -> "was done"
Line 218: "it has been done" -> "has been done"
Line 340: "feeder" -> "feeders"
Line 342: "dances" -> "danced"
Line 352: "Eavesdropping waggle dances" -> "Eavesdropping on waggle dances"
Lines 387-388: "Making an own calibration for the local bees is anyway recommended" -> I would suggest: "We also recommend generating a calibration specific to the bees of interest"
Experimental design: You clearly illustrated why it is important to know whether the waggle run duration-distance function is linear. You presented a thorough review of the literature on the subject, including helpful figures and tables to show comparisons between different studies. Figures 1, 2, and 4 were quite interesting, and figure 3 was nicely-formatted to highlight the different functions for waggle phase, return phase, and circuit durations. The methods section is clear and the methods seem appropriate.

I have two minor suggestions, although these changes are not necessary for publication. First, I would suggest saying something more explicit about optic flow likely experienced along the route to the feeders in this study. Of course, there is still uncertainty about the degree to which different landscapes affect the optic flow experienced by bees during their flights. Also, you did say that all feeders were along homogeneous terrain (a straight gravel road through beech forest). However, you might add a sentence to more specifically address the potential consequences for optic flow as some readers will wonder about it. Second, it would be helpful to include the R code that you used in the supplementary materials. It was very helpful that you included a csv of your data.
Validity of the findings: Your findings are interesting and potentially have significant consequences for basic science questions about the evolution of the waggle dance and practical questions about how to accurately map waggle dances. I think you did a good job of acknowledging the limitations of your study, considering the relatively small sample size, the fact that you could not examine differences in the duration-distance calibration among bees, the range of distances that you examined, and the possibility that these findings may not be true for all subspecies of Apis mellifera or species of Apis. However, as you illustrated nicely in your figures using the data from von Frisch and Jander (1957), there are multiple studies pointing to a similar conclusion. Your findings clearly indicate the importance of having more site- and population-specific calibration datasets that include greater distances.
Additional comments: Thank you for revisiting this question. It was clearly needed. I enjoyed reading the paper. You wrote a very thoughtful discussion of the implications of your results, and your figures are both helpful and elegant. I look forward to seeing further work on this question with other populations of bees and with an even greater range of distances.


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: HONEY BEES COMMUNICATE DISTANCE VIA NON-LINEAR WAGGLE DURATION FUNCTIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have went above and beyond in addressing my concerns in this section. The writing has improved and is far more clear.
Experimental design: They have addressed my primary concern regarding feeder conditions.
Validity of the findings: No comment.
Additional comments: I think the authors have done a great job in improving an already good paper. They met my concerns and went above my expectations in addressing them. Well done!


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: HONEY BEES COMMUNICATE DISTANCE VIA NON-LINEAR WAGGLE DURATION FUNCTIONS
Review round: 2
Reviewer: 2

Output:
Basic reporting: All comments were addressed by authors
Experimental design: All comments were addressed by authors
Validity of the findings: No comment
Additional comments: You did an excellent job of clarifying the text and methods, including adding references and sharing your R code, which is very helpful. It is an interesting study, and I look forward to seeing it published.


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

Input:
Title: CONSTRUCTION AND VALIDATION OF AN RNA-BINDING PROTEIN-ASSOCIATED PROGNOSTIC MODEL FOR COLORECTAL CANCER
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: Line 109: Please briefly describe the inclusion and exclusion criteria
Please add the IHC method under the Method Section
What’s your clinical trial registry?
Validity of the findings: Line 374: The database provided the association in a cross-sectional manner, which does not show causality. So the RBPs are just showing diagnostic/prognostic functions. Further studies are warranted to determine their therapeutic effects.

The authors have identified multiple RBPs that are correlated with CRC progression. Is there any RBP or couple of RBPs you want to highlight?
Additional comments: The authors provided novel and extensive evidence showing that several RBPs are differentially expressed in CRC vs non-CRC tissues, and could be potential biomarkers of CRC progression. The study was well-designed and correctly translated.


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: CONSTRUCTION AND VALIDATION OF AN RNA-BINDING PROTEIN-ASSOCIATED PROGNOSTIC MODEL FOR COLORECTAL CANCER
Review round: 1
Reviewer: 2

Output:
Basic reporting: This paper is well-organized with background material that can be improved. There can be several improvements that can be made to the manuscript to improve basic reporting and overall usefulness.

a) minor Grammar Issues
1) "shown that that" should be "shown that " on line 89
2) "are still poorly understand" on line 339 should be "are still poorly understood"
3) missing articles at some places throughout the paper.

b) Background context

1) "the role of RBPs in tumor development is relatively unknown" is pretty bold since there are multiple studies published over last few years

b) I think authors have skipped some background information about various studies done recently which studied the role of RBC's in CRC
Integrated analysis of RNA-binding proteins in human colorectal cancer | World Journal of Surgical Oncology | Full Text (biomedcentral.com)

c) Figures and tables

1) Authors should improve legibility of text and resolution in all figures except figure-4

2) Risk Score should be mathematically written
Experimental design: Experimental design
The article Fits well within the Aims and Scope of the journal.

1) Did the Authors perform some variable selection or regularization to reduce overfitting in the case of multivariate cox regression? Did they select a model just based on AIC criteria

2) Is there any rationale for selecting mentioned GSE studies as a similar methodology is used in different studies :
Integrated analysis of RNA-binding proteins in human colorectal cancer | World Journal of Surgical Oncology | Full Text (biomedcentral.com)
Frontiers | Molecular Characterization and Clinical Relevance of RNA Binding Proteins in Colorectal Cancer | Genetics (frontiersin.org)
Validity of the findings: No comments
Additional comments: No additional comments


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

Input:
Title: CONSTRUCTION AND VALIDATION OF AN RNA-BINDING PROTEIN-ASSOCIATED PROGNOSTIC MODEL FOR COLORECTAL CANCER
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The revised manuscript has been improved significantly.


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: CONSTRUCTION AND VALIDATION OF AN RNA-BINDING PROTEIN-ASSOCIATED PROGNOSTIC MODEL FOR COLORECTAL CANCER
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have addressed my comments
Experimental design: The authors have addressed my comments
Validity of the findings: The authors have addressed my comments
Additional comments: Thanks for citing those two papers. Can you please include how your method differs from the below two papers in your introduction section( not just the rebuttal)

1) Integrated analysis of RNA-binding proteins in human colorectal cancer | World Journal of Surgical Oncology | Full Text (biomedcentral.com)

2) Frontiers | Molecular Characterization and Clinical Relevance of RNA Binding Proteins in Colorectal Cancer | Genetics (frontiersin.org)


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: UTILIZING NETWORK PHARMACOLOGY AND MOLECULAR DOCKING TO EXPLORE THE UNDERLYING MECHANISM OF GUIZHI FULING WAN IN TREATING ENDOMETRIOSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Wang and Zhou et al conducted a nice in silico network biological study to elucidate the potential mode of action of Guizhi Fuling Wan in endometriosis.

The English of the manuscript is professional, but a little polishing may be needed.

The background and the importance of the topic in the introduction are established.

Please add the database citations, not just the links (Line 104, 105, 114).
Experimental design: I have a few major problems with the manuscript.

1. Both the BATMAN-TCM database and the TCMSP database are using predicted targets.
The manuscript considers these targets as the "real" targets of the TCM compounds. We cannot be sure about it. The authors must consider this. This is a major limitation of the manuscript.

2. The authors used a network biological technique to find the hub proteins between their targets. It resulted already well-known hubs e.g. p53, EGFR, EGF, AKT1. The authors miss mentioning the study biases and network analysis in the discussion.

3. The network figures need to be redrawn. I strongly suggest for the authors to read Marai et al visualisation guide about network figures.  https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1007244
In details:
Figure 2: The mol numbers do not contain information for the reader. Please use the name of the molecules.
Figure 4 and 5: Both networks do not contain information. I am against to add a network figure only for degree selection. Please use a force-directed layout or spring embedded layout for it. That should draw the pathways and molecular functions which are involved in the putative mode of action of GZFLW. The large betweenness and degree nodes can be highlighted in the network. Does the network have multiple components? Are the ingredients of GZFLW are acting on the same part of the network? The network can be coloured by the five ingredients. This way it can show which way each of the 5 ingredients acts. Those targets that are targeted by more than 1 ingredient can be coloured by an additional colour.
Figure 6 The authors tried to answer which compound act on which hub protein. The double circle layout does not help. I suggest instead two columns/rows the highest degree compound/protein at the top. This way the interactions will be a bit more clear. Also, the compound names need to be on the figure just like in Figure 2.
Figure 7 I am not satisfied with the layout. A non-circular layout would be better. It would show the interactions between pathways and targets better. The figure by itself does not contain extra information a table can be as good. But I have another underlying problem of this part.

4. For the GO and the KEGG enrichment analysis (DAVID/METASCAPE) the backgrounds are not mentioned. The problem with this is the following: Tha authors filtered proteins which are involved in endometriosis and which are drug targets. The first is done by the filtering to the disease. The second is based on the used databases. Any drug-target database will only have the druggable genome. The intersection of these two sets will be highly overlapping to the specific targets of GZFLW.
The problem in the sense of pathway enrichment analysis we will hardly get any specific result to GZFLW, it is possible it is coming just by the filtering criteria. (endometriosis, drugable genome) To see which pathways/molecular functions are specific for the compound combination I suggest using an enrichment analysis of all the endometriosis targets from BATMAN-TCM database and the TCMSP.

5. If possible I suggest having more thorough docking study for each target.

6. Please describe where are the oral bioactivity and drug-likeness are coming from, and explain why these cut-offs were chosen.
Validity of the findings: I have not much to add except winch I described on the experimental design. The limitations of the study need to be addressed.

The compounds and their putative targets are discussed in the discussion. However, the targets are putative and this needs to be emphasised.
Additional comments: Please add a space before the citations.


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: UTILIZING NETWORK PHARMACOLOGY AND MOLECULAR DOCKING TO EXPLORE THE UNDERLYING MECHANISM OF GUIZHI FULING WAN IN TREATING ENDOMETRIOSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1. The introduction needs more details. It is suggested that the authors improve the description at lines 71- 81 to make the introduction of Traditional Chinese medicine (TCM) network pharmacology clear and have literature well referenced.
2. In addition, to improve the readability, the language of this manuscript should be improved, e.g., the meaning of "multi-component target medicine" is ambiguous in Page 3.
Experimental design: There are some unclear points described in Method section.
1.	The collection of GZFLW ingredients should consider some other high-quality databases, then the results from different databases should be sufficiently compared to obtain more complete and reliable GZFLW ingredients.
2.	The GZFLW targets prediction in this manuscript is rather like targets collection. How to ensure the reliability of GZFLW targets for further study should be sufficiently considered.
3.	In addition, the definition of “Betweenness Centrality” (Line 120) should be checked again.
4.    How to condiser the traditional efficacy of GZFLW in treating endometriosis? It is suggested that authors take into consideration of the traditional efficacy of GZFLW based on the biological network related to TCM syndrome (ZHENG), for example, the Cold Syndrome network or the Hot Syndrome network (IET Syst Biol 2007 [PMID: 17370429]).
Validity of the findings: 1.	In discussion section, the expression of “immune response” and “Inflammatory response” should be unified.
2.	It is not enough to just list the analysis results of GO and Pathway enrichment and infer that combined synergetic effects of these pathways were the mechanisms of GZFLW. It should be provided with several aspects of solid evidence to support these results.
Additional comments: I suggest authors make major revisions of this paper.


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

Input:
Title: UTILIZING NETWORK PHARMACOLOGY AND MOLECULAR DOCKING TO EXPLORE THE UNDERLYING MECHANISM OF GUIZHI FULING WAN IN TREATING ENDOMETRIOSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript improved. I am really happy that the authors have addressed the network visualisation comments of mine. The network figures look much better.  However, I still have some points which need to be addressed, mostly English.

I will list here some English mistakes which need to be addressed:
Line 55 extra space
Line 56 extra space
Line 70-71 use instead of "to network target mediated by a multi-component system"  use "to multiple interacting targets mediated by multiple compounds."
Line 78 I have never heard the word "systematicness"
Line 84 through the STRING database
Line 118 The active compounds' targets in GZFLW were obtained...
Line 127 The interactions of the targets were obtained, not the targets.
Line 131  Betweenness centrality is the number of the shortest path going through a given node/edge. Sometimes it is measured by the absolute value or the percentage of all shortest paths. - The authors use percentage-based betweenness centrality in their manuscript.
Line 156 KEGG citation missing
Line 157 Cytoscape citation missing
Line 196 performed and resulted in 116 GO...
Line 249 I guess: The differentially expressed genes are related to inflammation in EM.?
Line 287-290 The authors are using "tight combination" instead of high docking score or strong putative interaction. Please use one of the tight combinations is meaningless in this case.

Figure 2 Please add the figure description of the ingredients name. It makes it much easier to read.

Table 2 The Betweenness centrality is not in the table
Table 3 What is the measurement of the docking score? Usually, it is some kind of energy best of my knowledge?
Experimental design: What was the cut-off for "tight combination"?
Validity of the findings: I have here two large issues.

The authors forget to mention after I have asked them in my previous revision that most of the targets of the compounds are predicted, putative targets. In their response, they mention that they wrote in the limitation paragraph, but it is not there.

Another validity question regarding the study is the interaction of the compounds and various ligands such as EGF, VEGEFA, IL-6. It is hard for me to imagine biologically relevant inhibition or effect if the compound directly targets the ligands and not the receptors. The authors specifically mention EGF as their most strongly interacting protein with the TCM ingredients target. Why do they think that the interaction is relevant? What do the TCM compounds do with EGF? Use less strong language, if no further validity is available.
Additional comments: The manuscript improved a lot. Please mention that the databases are mostly predicted interactions! Even after the docking study, the results are suggesting a putative mechanism of action of the TCM combination. It is a nice network biological work but this is the maximum what we can say based on the 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: UTILIZING NETWORK PHARMACOLOGY AND MOLECULAR DOCKING TO EXPLORE THE UNDERLYING MECHANISM OF GUIZHI FULING WAN IN TREATING ENDOMETRIOSIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: In the revision, authors did not provide any solid and new evidence from experimental and clinical studies to ensure the reliability of GZFLW targets, GO and Pathways, as well as synergetic effects to support their claims.
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: OVEREXPRESSION OF A ROSA RUGOSA THUNB. NUDX GENE ENHANCES BIOSYNTHESIS OF SCENT VOLATILES IN PETUNIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comments
Experimental design: No comments
Validity of the findings: No comments
Additional comments: The article is devoted to the study of  petunia plants transformed with the NUDX gene from rosa plants.
The authors believe that this gene is capable of increasing the content of aromatic oils and is promising for biotechnology.
The authors have done a great work, obtained important data of interest for modern plant physiology.
The experiment was well thought out, the set goals were achieved, and adequate methods were applied.
However, there are minor comments on the work.

One of the main remarks is the lack of a description of the statistical methods used by the authors in the article.

In the materials and methods, it is necessary to write in more detail how and under what conditions calli were formed, how the regeneration was carried out, to describe in more detail the conditions of electroporation and what device was used for this.

I'm not a native speaker and it's hard for me to judge, but there are a few mistakes in 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: OVEREXPRESSION OF A ROSA RUGOSA THUNB. NUDX GENE ENHANCES BIOSYNTHESIS OF SCENT VOLATILES IN PETUNIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The basic reporting of the manuscript entitled 'Overexpresstion of a Rosa rugosa NUDX gene enhances biosynthesis of scent volatiles in petunia' is sound and concise. The manuscript reads well and has a great flow. The authors covered a wide range of literature in the intro. However, there are a few grammatical errors that have been highlighted in the attached document.
Experimental design: The experimental design is non existent. I have highlighted the suggestions on how to include the experimental design in the manuscript. Check the attached review document for more explanation.
Validity of the findings: This study covered a wide range of aspects from gene expression, to metabolomics, physiological and the extend of the results obtained are valid and coherent. The biological pathways investigated corresponded with the results obtained.
Additional comments: In general, the manuscript is well written and contains sound results. However, the discussion section lacks depth. I suggest that the authors validate their findings by explaining the biological processes obtained with some literature. In addition, the authors also need to compare and contrast their findings with previous studies. Further suggestions are included in the attached document.


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: OVEREXPRESSION OF A ROSA RUGOSA THUNB. NUDX GENE ENHANCES BIOSYNTHESIS OF SCENT VOLATILES IN PETUNIA
Review round: 1
Reviewer: 3

Output:
Basic reporting: In this manuscript author's have nicely compiled their hypothesis and results to show that NUDX1 gene can be utilized for enhancing the scent of flowers in different flower species.

English need some improvement otherwise literature is cited in sufficient detail. Rest of the structure of the manuscript is fine.
Experimental design: Experimental design is well planned. Research question has approached the practical aspect of increasing demand of aromas.
Validity of the findings: The functions of the gene NUDX1 is validated  in petunia to confirm the hypothesis. Conclusion and inferences made in the discussion are well supported by the results.
Additional comments: 1. From the introduction what I understood that NUDX are multiple genes then what is special about targeting the NUDX1 gene? Is this based on homology with Arabidopsis or based on the information available in Magnard et al., 2015? Explaining it in introduction is important because whole story of the paper is around NUDX1.

2. Line 30: Consider using “expensive” in place of extremely expensive.

3. Line 37: Revise the sentence “Terpenes are the most abundant volatiles in floral aroma”, including…..

4. Line 153: Add citation for the software or from where you get this software.

5. Fig 2: For P2 tissue the expression is low or comparable with other remaining low expression tissue P3 but the content of citonellol is quite high. For this tissue expression level may not be consistent with content.

6. Applying statistical significance test on the expression and metabolite content is important.


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

Input:
Title: A TOTAL OF 219 METAGENOME-ASSEMBLED GENOMES OF MICROORGANISMS FROM ICELANDIC MARINE WATERS
Review round: 1
Reviewer: 1

Output:
Basic reporting: In their study, Jégousse and co-workers describe 219 archaeal and bacterial MAGs reconstructed from 31 metagenomes taken close to Iceland. The manuscript is well organized and written. It contains most of the information necessary for this type of article (see details below). The figures are appealing. The raw data has been deposited in a public repository.
Experimental design: Te manuscript is a data report. The methods section is fine but the information about the co assembly and the binning is missing (You need to check the figure and the results to llearn which tools were used). In addition, the authors should provide version numbers for all tools used (MegaHIT, CheckM, MetaBAT2, CONCOOT, MaxBin2, prodigal and so on). Also provide the version of the GTDB database.
Validity of the findings: All data has been provided with one exception. Please upload your MAGs to GenBank (the fasta file for each MAG). Otherwise this important information is not available to researchers.
Additional comments: Good Job. Only my previous comments and three minor comments:

Line 18: ... and sink lower depths of the ocean. -> Rephrase.
Line 23: (please change to) ... to bridge this knowledge gap.
Line 49: uncultured microorganisms (change to) -> yet uncultured marine lineages